1. No category

Anatomy and
Neuroscience
Research Projects for 2013
(Includes Honours and PhD)
Research in the Department of Anatomy and Neuroscience
There are many opportunities for students to undertake research in the Department of Anatomy and
Neuroscience. An Honours year is open to students who have completed an undergraduate degree,
while students with a BSc (Hons) or BBiomed (Hons) can train for a career in science by undertaking a
three to four year PhD.
In this brochure, you will find a wide range of research projects that can meet the needs of potential
Honours or PhD students. You should look through the projects and contact the Head of the laboratory
of any projects that interest you. Alternatively, you can contact the departmental co-ordinator for
Honours and PhD, Dr Peter Kitchener (8344 6746, [email protected]).
CONTENTS
Honours ..............................................................................................................................................5
PhD ......................................................................................................................................................6
Biomechanical Engineering (David Ackland)
Project 1: Single leg squat performance in runners: biomechanical analysis and orthotic
effects ............................................................................................................................................7
Project 2: Strength of a novel shoulder rotator cuff reconstruction technique .............................. 7
Neuronal Differentiation Laboratory (Colin Anderson)
Determining neuronal identity during development....................................................................... 8
Cell Migration Laboratory (Richard Anderson)
Project 1: Factors influencing the migration of neural crest cells within the caecum. .................. 9
Project 2: Role of L1CAM as a modifier gene in Hirschsprung’s disease. ................................. 10
Ocular Development Laboratory (Robb de Iongh)
Project 1: Role of integrin-linked kinase in lens development and cataract ............................... 11
Project 2: Role of Fz5 and Fz7 genes in lens development ....................................................... 11
Stem Cell Laboratory (Mirella Dottori, Mark Denham)
Project 1: Investigating specification of neural progenitors using human pluripotent stem cells12
Project 2: Characterizing Neural Progenitors Derived from Friedreich Ataxia Induced
pluripotent stem cells ...................................................................................................................12
Project 3: Generation of dopaminergic neural progenitor cells from human pluripotent stem
cells ..............................................................................................................................................12
Visual Neuroscience Laboratory (Erica Fletcher, Una Greferath)
Project 1: Investigation of potential therapeutic targets aimed at inhibiting photoreceptor death
in a mouse model of retinal degeneration ...................................................................................13
Project 2: The role of microglia in Age-Related Macular Degeneration ..................................... 14
Project 3: The role of histamine in the development of the retina .............................................. 14
Autonomic and Sensory Neuroscience Laboratory (John Furness, James Brock)
Digestive Health and the Enteric Nervous System: From Molecule to Human ......................... 15
Transplantation science: Improving the survival of transplanted organs................................... 15
Nutrition: How nutrients modify digestive function ...................................................................... 15
Spinal Cord Injury: Failure of bladder, bowel and blood pressure control ................................. 16
Finding novel receptors important for breast and other cancers ................................................ 16
Mechanisms regulating neural control of arterial vessel constriction ......................................... 16
Structure function relationships in peripheral sensory nerve terminals ...................................... 16
Neuron Development and Plasticity Laboratory (Jenny Gunnersen)
Molecular mechanisms of maladaptive plasticity in neuropathic pain ........................................ 17
Is Sez-6 important for learning and memory in the adult brain? ................................................. 18
Secreted signalling molecules in excitatory synapse development ............................................ 18
The role of TCF7L2 (TCF4) in regeneration after spinal cord injury ........................................... 18
Stem Cell Genetics Laboratory (Gary Hime)
Analysing the role of transcriptional repressors in Drosophila and mouse stem cells ............... 19
Analysing genetic roles of translational repressors in Drosophila stem cells ............................. 20
Analysing the role of a novel zinc finger protein in stem cell maintenance ................................ 20
How does regulation of nuclear importation affect stem cell maintenance ................................ 20
Front cover image by Annette Bergner, Department of Anatomy and Neuroscience
2
Neural Mechanisms of Bone Pain (Jason Ivanusic)
Project 1: Changes in the neuro-chemical phenotype of primary afferent neurons that innervate
bone in an animal model of inflammatory bone pain. ................................................................. 21
Project 2: Changes in neurotrophin signalling molecules in primary afferent neurons that
innervate bone in an animal model of inflammatory bone pain .................................................. 21
Project 3: Investigation of the central nervous system areas associated with bone pain .......... 22
Neural Development, Injury and Pain Laboratory (Janet Keast, Peregrine Osborne)
Project 1: Development of sacral autonomic circuits .................................................................. 23
Project 2: Improving the recovery of injured visceral nerves ...................................................... 23
Project 3: How nociceptor neurons change in a rodent model of endometriosis ....................... 24
Project 4: Neuroanatomical specialisation of pelvic sensory and pain pathways....................... 24
Project 5: Neuropharmacology of the GDNF family of neurotrophic factors in peripheral nerve
regeneration and pain..................................................................................................................24
Project 6: Mechanisms of central pain after spinal cord injury (SCI) .......................................... 24
Project 7: New uses of local anaesthetics in the treatment of visceral pain ............................... 25
Project 8: Application of multielectrode arrays (MEAs) in models of chronic pain ..................... 25
Project 9: Sensory functions of the bladder urothelium in a co-culture system .......................... 25
Glia, Myelin Biology and Multiple Sclerosis Research (Trevor Kilpatrick)
Major Project Areas, Overview ..........................................................................................................26
CNS Myelination Laboratory (Ben Emery) ........................................................................................27
Projects available ........................................................................................................................27
Myelin Repair Laboratory (Holly Cate) ..............................................................................................28
Projects available ........................................................................................................................28
Neurotrophin Signaling Laboratory (Simon Murray, Junhua Xiao) ................................................... 29
The influence of BDNF on CNS & PNS Myelination ................................................................... 29
Multiple Sclerosis Genetics Laboratory (Judith Field) ....................................................................... 30
Regulation of Interleukin 2 receptor alpha (IL2RA) DNA methylation in MS .............................. 30
Functional consequences of Multiple Sclerosis risk genes......................................................... 30
Early Brain Development Laboratory (Peter Kitchener)
Project 1: Microglial invasion and migration into the embryonic brain........................................ 31
Project 2: The formation of the cardiovascular control centres in brainstem.............................. 31
Behavioural Neuroscience at the Florey (Andrew Lawrence)
Project 1: The effects of toluene exposure on the adolescent brain........................................... 32
Project 2: Identification of hypothalamic peptides that regulate reward-seeking........................ 33
Project 3: Neural circuitry underlying extinction of fear across development ............................. 33
Project 4: Extinction of drug-seeking: pathways & mechanisms ................................................ 34
Brain and Behaviour Laboratory (Mark Murphy)
Project 1: What circuits in the brain are involved in a particular behaviour? .............................. 35
Project 2: Plasticity in learning and memory. .............................................................................. 35
Project 3: Genetics of Behaviour.................................................................................................35
Project 4: Methods to ameliorate cognitive decline and memory deficits in Alzheimer’s
disease ........................................................................................................................................36
Neurotrophin Signaling Laboratory (Simon Murray)
Project 1: Analysis of BDNF and its mimetics ............................................................................. 37
Project 2: Mechanisms of growth factor signaling....................................................................... 38
Animal Brain Imaging using MRI (Roger Ordidge)
Project 1: Optimisation of MRI tractography by Diffusion-weighted MRI .................................... 39
The Ion Channels and Disease Laboratory (Steven Petrou)
Project 1: Multi site patch clamp recording of cortical micro networks ....................................... 40
Project 2: High density multi-electrode array recording of in vitro networks in epilepsy ............ 40
Project 3: In vivo electrophysiological analysis in mouse models of genetic epilepsy ............... 40
Project 4: The glass brain: "Connectomics" in epilepsy .............................................................. 41
Project 5: MRI tractography in mouse models of genetic epilepsy: Creation of prognostic and
diagnostic structural biomarkers..................................................................................................41
3
The Ion Channels and Disease Laboratory (continued)
Project 6: High content automated analysis of ion channels in epilepsy .................................... 41
Project 7: Optogenetic modulation of the area tempestas - an epilepsy hot spot ...................... 41
Project 8: Exploring the role of GABA mediated tonic inhibition in depression .......................... 42
Project 9: In vitro study of the mechanism of action of a naturally occurring pain killer ............. 42
Project 10: Zinc and seizures ......................................................................................................42
Project 11: HCN channels, epilepsy and memory ...................................................................... 42
Physical Anthropology Laboratory (Varsha Pilbrow)
PhD in ancient DNA.....................................................................................................................43
MSc in isotope analysis ...............................................................................................................43
MSc in discrete dental trait analysis ............................................................................................43
Honours in hominid taxonomy .....................................................................................................44
Animal Models for Cancer and Neurodegenerative Disease (Leonie Quinn)
Project 1: Transcriptional control of the MYC oncogene ............................................................ 45
Project 2: Drosophila models for leukemia .................................................................................45
Project 3: Drosophila models for neurodegenerative disease .................................................... 46
Project 4: Dissecting growth regulation by novel signaling pathways ........................................ 46
Project 5: The MYC oncogene in stem cells ............................................................................... 46
Project 6: Defining novel roles for ASCIZ....................................................................................46
Neural Regeneration Laboratory (Ann Turnley, Kim Christie)
Project 1: Does traumatic brain injury (TBI) regulate hypothalamic neurogenesis? .................. 47
Cancer Biology Laboratory (Elizabeth Vincan)
The FZD7/Wnt Signalling Pathway in Hepatocellular Carcinoma .............................................. 48
Autonomic Neuron Development Laboratory (Heather Young)
Project 1: Live cell imaging of neural crest cell migration along the developing gut .................. 49
Project 2: Migration of melanoma cells in the neural crest environment .................................... 49
Project 3: Proliferation of enteric neuron precursors................................................................... 50
Project 4: Potential of cell therapy to treat Hirschsprung’s disease............................................ 50
4
HONOURS
What is a BSc (Hons) or BBiomed (Hons)?
A BSc (Hons) or BBiomed (Hons) is a year of study following a Bachelor of Science or Bachelor of
Biomedicine degree. It consists of a combination of a research project and course work. The research
component is carried out under supervision in one of the research groups in the department. Your
research project is included in the 75 point subject "Anatomy and Neuroscience Research Project".
80% of this subject is assessed via 7000 word research thesis submitted at the end of the year. Other
assessment tasks include a literature review, a short oral presentation of your work as well as a grade
from your supervisor. The remaining 25 points are from two 12.5 point coursework subjects,
“Introduction to Biomedical Research” and “Seminars in Anatomy and Neuroscience”.
The BSc (Hons) or BBiomed (Hons) year starts in February and ends in November and is only available
full time.
Entry requirements
You must have completed a suitable degree (B.Sc., B.Biomed. or equivalent) and achieved a faculty
honours score of 65 or equivalent. You must also have the agreement of a supervisor in the Department
of Anatomy and Neuroscience to supervise you in the project.
How to Apply
Detailed instructions on how to apply, and the circuital dates, are available at the MDHS student centre
website:
http://www.sc.mdhs.unimelb.edu.au/how-apply
Essentially, there are three sequential steps in applying for honours.
Step 1: You will need to decide which projects / supervisor you would like to do your honours year with:
because there may be more applicants than places available for particular projects or supervisors, it is
important to identify a number of potential projects (within this department or other departments in the
MDHS Faculty). Information about the range of honours research projects is provided on our
departmental website, and also in hardcopy (this booklet). In addition, it is a good idea to come to the
department’s Honours Information Session, which is held mid September (details of the time and
location will be advertised on our website from late August). This session is an opportunity to meet and
talk with supervisors from all of the research laboratories in the department. Information about projects
in this department, and other departments in the Faculty, will also be available at the MDHS faculty
honours information session (also held in mid September).
Step 2: All applicants (local and International) must lodge an online application for Honours to the
Faculty of Science. (See the website above for instructions and timelines.)
Step 3: After having decided on a project(s) and submitting your online application, you will need to
lodge your project preferences with MDHS through the Honours Application and Tracking System
(HATS). It is essential that you have already identified which projects you wish to apply for by speaking
to potential supervisors (i.e. Step 1) and have applied for Honours through the Student Portal (i.e. Step
2) before you carry out Step 3.
5
PhD
What is a PhD?
A PhD is three to four years full time work doing supervised research in a laboratory. It is a necessary
step in a career path to becoming a professional scientist. There are no exams or course work. The
research work is written up as a thesis and assessed by two experts in the field. By the end of a PhD,
you will be able to work as a professional scientist.
Why do a PhD in the Department of Anatomy and Neuroscience?
The Department of Anatomy and Neuroscience has long been a leading research department in the
Faculty of Medicine, Dentistry and Health Sciences. We offer a range of research topics in anatomy,
neuroscience, cell and developmental biology and molecular biology, and many of the laboratories
collaborate with leading research groups around the world. The dynamic research in the department is
due in part to the excellent facilities, as well as to the presence of a range of full time research-only
staff.
Where does a PhD lead?
Following completion of a PhD in Anatomy and Neuroscience, you might move on to a job, often
overseas, in a research institute or University, working full time as a scientist. You could remain working
full time as a Research Scientist for the rest of your working life or eventually take a job as an academic
in a University, doing both research and teaching. A PhD in Anatomy and Neuroscience may also lead
to a range of other jobs, particularly if you have other postgraduate qualifications. These jobs could
include patent attorney, clinical trials or research co-ordinator, or biotechnology manager.
What to do if you are interested?
The first step is to identify projects of interest from this booklet and contact a potential supervisor, who
will assist you with the enrolment process. For general information about postgraduate research in the
department, contact the PhD Co-ordinator, Dr Peter Kitchener.
Entry requirements
Entry requirements to a PhD are that you have a BSc (Hons), BBiomed (Hons) or equivalent degree.
PhD scholarships are also available to support you while you study. They are competitively awarded on
the basis of your academic record.
Further information can be found at http://www.anatomy.unimelb.edu.au/researchprojects/phd.html.
6
Biomechanical Engineering
Dr Natalie Collins (Room 207, Level 2, Mechanical Engineering, ph. 8344 3910)
Email: [email protected]
Project 1: Single leg squat performance in runners: biomechanical analysis
and orthotic effects
The single leg squat is a simple clinical tool used by physiotherapists
and other health professionals to evaluate dynamic unilateral lower
limb control. Patients are rated as “good” or “poor” based on specific
observed criteria that relate to trunk, pelvic, hip, knee and foot
control, as well as overall performance. This study will investigate
biomechanical analysis of the single leg squat task in order to
address three specific aims. The primary aim is to determine
whether runners who have been rated as “poor” demonstrate
different biomechanics (kinematics and kinetics) during the single leg
squat compared to runners rated as “good”. Secondly, it will
investigate whether biomechanical performance correlates with
clinical measures of balance, strength, and lower limb alignment.
The third aim is to evaluate whether off-the-shelf foot orthoses can
improve biomechanics during the single leg squat in those who have
been rated as “poor”.
Dr David Ackland (Room 205, Level 2, Mechanical Engineering, ph. 8344 8646)
Email: [email protected]
Assoc Prof Martin Richardson
Email: [email protected]
Project 2: Strength of a novel shoulder rotator cuff reconstruction technique
Rotator cuff tears are a common cause of shoulder pain and account for approximately 75,000
operations each year. The past decade has seen the evolution of rotator cuff tear management from
open and minimally open repairs, to all-arthroscopic techniques. The now common use of suture
anchors in shoulder surgery has revolutionised athroscopic rotator cuff repair. Despite technological
advances, complications may occur with arthroscopic suture anchors, including failure of the tissue,
suture, or anchor before healing has occurred. This project, in conjunction with the orthopedic company
Smith & Nephew, will investigate the pull-out strength of a novel anchor and double row suture
combination. It is hypothesised that using a conventional suture anchor in combination with a double
row of sutures will lead to improved tendon fixation compared to the anchor alone or the anchor in
combination with a single row of sutures. Experiments will be performed on shoulder cadavers using a
material test system (MTS) and custom designed fixtures. The student will work with an orthopedic
surgeon to perform the shoulder reconstructions.
7
Neuronal Differentiation Laboratory
Determining neuronal identity during development.
Assoc Prof Colin Anderson (Room E723, ph 8344 5807)
Email: [email protected]
Dr Kylie Cane (Room E503, ph 8344 3979)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/anderson/index.html
The nervous system is the most complex of all biological structures. It contains many different types of
neurons, which differ in the neurotransmitter they release, the receptors they express and the
connections that they make. Yet many neurons that are distinctly different in their mature form originate
from the same stem cells. Furthermore, this process yields the right number of cells in the right place
and at the right time to create a functional network of neurons. Understanding the processes that control
expression of neuronal diversity is one of the most challenging problems facing biologists today.
In our laboratory, we study autonomic neurons to determine how they adopt their distinctive, mature
forms.
1. How do neurons and glial cells arise from a common pool of neural crest progenitors in a common
environment?
2. What is the relationship between when a neuron is born and the phenotype it later adopts?
3. Does the developing sympathetic ganglion exhibit any topography in the distribution of stem cells and
neuronal and glial precursors?
4. How is the axon of a developing autonomic neuron directed to its target?
Our approach is to use mice as models. This allows us to use a range of transgenic animals, with
specific genes inactivated or with reporter genes indicating when specific genes are activated. In
combination with modern culture and time lapse techniques, this gives us a powerful insight into
development in a mammalian embryo.
Any one of these questions would make a suitable topic for a PhD, Honours, AMS or 516307 project.
Projects can also be tailored to match the specific interests or goals of a particular student. All projects
will provide training in key techniques widely used in modern biological sciences, Including multiplelabelling immunofluorescence, organ culture and confocal microscopy and will provide the basis for a
range of future career choices. Students should feel free to contact me to discuss possible projects in
my laboratory, or to discuss Honours in general.
8
Cell Migration Laboratory
Dr Richard Anderson (Room E525, ph 8344 5783)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/anderson_r/index.html
During the development of the enteric nervous system (ENS), neural precursors from the hindbrain
must first migrate into and colonise the entire gastrointestinal tract (gut). This migration is very
interesting because (a) it takes a long time (>4 weeks in humans and 4 days in mice), (b) the cells have
to migrate very long distances, particularly those that colonise the colon and rectum, and (c) if cells fail
to colonize the distal gut in humans, a disease called Hirschsprung’s disease results which requires
surgery.
Timetable of enteric neural crest cell migration
Project 1: Factors influencing the migration of neural crest cells within the
caecum.
We have recently shown that when neural crest cells reach the caecum, their migratory properties are
altered such that they are no longer able to migrate within the midgut. This suggests that the caecum
possess molecules that can alter neural crest cell migration. In this project, you will identify molecules
that are expressed within the caecum (using microarrays and in situ hybridisation) and then examine
whether these molecules alter cell migration (using a variety of in vitro assays).
Comparison of distance that pre-caecal and caecal cells migrate in recipient precaecal and post-caecal gut. The most distal cell is indicated with an open arrow.
Pre-caecal cells colonised both pre-caecal and post-caecal recipient gut equally
well (top panel). Caecal cells colonised pre-caecal recipient explants poorly, but
migrated a similar distance along post-caecal recipients to pre-caecal cells
(bottom panel).
9
Project 2: Role of L1CAM as a modifier gene in Hirschsprung’s disease.
L1CAM is a X-linked gene that encodes the cell adhesion molecule, L1. In humans, mutations in
L1CAM have been implicated in a range of neurological disorders. Notably, some individuals with
L1CAM mutations also have Hirschsprung’s disease, suggesting a possible role for this gene in ENS
development. We have recently shown that: (i) L1 is expressed by enteric neural crest cells as they
migrate through the developing mouse gut; (ii) disrupting L1 activity retards enteric neural crest cell
migration in explants of embryonic mouse gut in vitro; and (iii) L1CAM null mutant mice show a
significant delay in enteric neural crest cell migration. However, the entire gastrointestinal tract is fully
colonised prior to birth. This suggests that L1CAM may function as a X-linked modifier gene in
Hirschsprung’s disease. A modifier gene is defined as a gene that when mutated, is insufficient on its
own to produce an effect. However, when coupled with another genetic mutation, it produces or
enhances an effect. In this project, you will examine whether genetic interactions between L1CAM and
other known Hirschsprung susceptibility genes results in a Hirschsprung-like phenotype. To do this you
will use a variety of methods, including genetic and molecular analysis, microdissection, histochemistry
and confocal microscopy.
Wholemount
preparation
of
gut
from
+/+
+/-/+/L1CAM ;Sox10 (A, C) and L1CAM ;Sox10
(B, D) mice.
A megacolon is apparent
-/+/macroscopically in the L1CAM ;Sox10 gut
+
(B). (C-D) At higher power, β-gal neurons are
present in ganglia (arrowheads) within the distal
most region of the large intestine of
+/+
+/L1CAM ;Sox10 mice (C), but are absent
-/+/from the gut of L1CAM ;Sox10 mice (D).
10
Ocular Development Laboratory
Dr Robb de Iongh (Room E631, ph 8344 5788)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/deIongh/index.html
Overview
Research in the Ocular Development Laboratory is directed at identifying the molecular mechanisms
that regulate development, growth and pathology of the eye, particularly the lens. We use a variety of
techniques to investigate the genes and signalling pathways involved in the cellular interactions and
processes that control the development, structure and growth of the eye in the vertebrate embryo.
Project 1: Role of integrin-linked kinase in lens development and cataract.
During development of the vertebrate lens there are dynamic interactions between the extracellular
matrix (ECM) of the lens capsule and lens cells. Disruption of these interactions can result in
perturbation of lens development and cataract. Integrins are key receptors for ECM signals and various
studies have documented distinct repertoires of integrin expression during lens development, and in
anterior subcapsular cataract (ASC). One of the key mediators of integrin signalling in the cell
cytoplasm is integrin-linked kinase. We have generated mice with conditional null mutation of Ilk and
found that it affects cell proliferation and survival. However, our, cell culture experiments indicate that
over-activation of ILK can lead to an epithelial-mesenchymal transition (EMT), similar to that found in a
form of human cataract. We have now generated transgenic mice that express a hyperactive kinase
form of Ilk in the lens. This transgene can rescue the null phenotype and shows effects on lens
epithelial cell morphology in vitro.
This project will further investigate the effects of the hyperactive Ilk transgene on lens cell phenotype.
Techniques will include management of mouse colonies, PCR genotyping, and examining the
responsiveness of epithelial cells to TGFβ and FGF in tissue culture and examining molecular markers
by RT-PCR, immuno-histochemistry/western blotting and in situ hybridisation. The project may also
involve generating a new transgene for a kinase-dead form of Ilk, which will be used to generate a
transgenic mouse line.
Project 2: Role of Fz5 and Fz7 genes in lens development.
We have already shown that the Wnt/β-catenin pathway is central to regulating lens stem and
1
progenitor cells during development. Recent evidence indicates that the Wnt/PCP pathway also plays
2
roles in lens development. To investigate the contribution of Frizzled (Fz) receptors in mediating Wnt
signals we are conducting Cre-LoxP experiments to conditionally delete Fz5 and Fz7 genes from the
developing lens. Techniques include breeding and management of mouse colonies, histology, and
investigating molecular markers of lens development by RT-PCR, immuno-histochemistry/western
blotting and in situ hybridisation.
1
Cain S et al., Dev Biol 321:420 (2008); Martinez G et al., Invest Ophthalmol Vis Sci 50:4794
(2009).
2
Sugiyama Y et al., Dev Biol 338:193 (2010).
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Stem Cell Laboratory
Dr Mirella Dottori ([email protected])
Dr Mark Denham ([email protected])
Level 2, Kenneth Myer Building, ph. 8344 3988
Project 1: Investigating specification of neural progenitors using human
pluripotent stem cells.
Human pluripotent stem cells serve as a useful cellular model to study early patterning and cell fate
events occurring during human embryogenesis. We have recently developed a chemically defined
system that enables us to generate neural crest cells from human pluripotent stem cells. During
embryogenesis, neural crest cells generate most of the peripheral nervous system but they also
differentiate to non-neural cell types, including cartilage and melanocytes. Lineage determination of
neural crest progenitors depends on spatial and temporal cues occurring during development. This
project is aimed at identifying the extrinsic and intrinsic signals that determine commitment of human
stem cell-derived neural crest progenitors to neural and non-neural lineages. These studies involve
culturing neural progenitors with various factors at different time points and then identifying their
potential to generate peripheral neurons and non-neuronal lineages. Techniques associated with this
project include tissue culture, immunofluorescence analyses and Q-PCR analyses.
Project 2: Characterizing Neural Progenitors Derived from Friedreich Ataxia
Induced pluripotent stem cells.
Friedreich ataxia (FA) is an autosomal recessive disease characterised by neurodegeneration and
cardiomyopathy and is the most common form of all inherited ataxias known to date. The cause of FA is
due to the presence of a trinucleotide GAA repeat expansion in the first intron of the FXN gene,
resulting in an insufficiency of the mitochondrial protein, Frataxin. Reduced levels of Frataxin protein
leads to mitochondrial dysfunction, cell toxicity and cell death, particularly within the nervous system
and cardiac tissue. In FA research there is a strong need to develop human cellular models of the
disease to further study the cellular pathology of FA as well as develop therapies. To meet these needs,
we have generated induced pluripotent stem cell lines derived from skin biopsy samples of FA patients.
We have shown that the FA iPS cells retain some of the fundamental molecular and genetic
characteristics of this disease, including low levels of Frataxin and GAA repeat instability within the FXN
gene locus. The main focus of this research project is directed at using the FA iPS cell lines to generate
neuronal cell types that predominantly degenerate in FA, and to examine how these cells may display
the disease phenotype. These studies are the first steps to obtaining a human cellular model system of
FA.
Project 3: Generation of dopaminergic neural progenitor cells from human
pluripotent stem cells
Parkinson’s Disease has been one of the major targets for using stem cells as a potential form of
treatment. This is because it is hypothesized that only a few dopaminergic neurons are required to
restore function. One of the major objectives of using stem cell therapies to treat Parkinson’s Disease is
to obtain human mesencephalic dopaminergic (mesDA) neural progenitors that, upon transplantation,
can integrate within the brain and restore function. hESC provide an unlimited source of stem cells that
can be used to investigate such objectives. During embryonic development, mesDA progenitors arise
from the floorplate regions of the neural tube. This floorplate expresses several transcription factors,
including Gli1, and FoxA2, all of which are required for mesDA neuronal fate. We are currently
optimizing how to activate intrinsic cellular pathways needed for generating mesDA neurons from
pluripotent stem cells. This approach is also useful for the generation of other ventral neural cell types
including motor neurons which are affected in Motor Neuron Disease. These studies will shape our
understanding of how to use stem cells to derive the most ideal donor cell type for transplantation
therapies and obtain the desirable outcome of restoring function.
12
Visual Neuroscience Laboratory
Dr Erica Fletcher (Room E721, ph 8344 3218)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/fletcher/index.html
Dr Kirstan Vessey (Room E708A, ph 8344 5769)
Email: [email protected]
Dr Andrew Jobling (Room E708A, ph 8344 5769)
Email: [email protected]
Dr Una Greferath (Room E702E / E708A, ph 8344 4315 / 8344 5769)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/greferath/index.html
Retinal diseases are a major cause of blindness in the Western world. There are few treatments
currently available, largely because the underlying mechanisms of disease are not well understood. Our
laboratory investigates these underlying disease mechanisms. We are currently studying two broad
classes of retinal diseases:
RETINAL DEGENERATIONS:
Death of the light-detecting retinal neurons, the photoreceptors, are associated with 50% of all cases of
blindness in Australia, being a major contributor to visual impairment in Age-Related Macular
Degeneration (AMD), and hereditary retinal degenerations including Retinitis Pigmentosa (RP). There
are currently no treatments for RP or AMD. We are examining the mechanisms of photoreceptor death
and whether specific treatments ameliorate or slow the loss of photoreceptors.
RETINAL VASCULAR DISEASE:
Retinopathy of Prematurity (ROP) is a major cause of visual impairment in children born prematurely.
ROP is a vascular disease, caused by excessive growth of blood vessels on the surface of the retina in
response to the combined effects of extreme immaturity of the retina and high levels of oxygen used for
critical care of neonates. Currently, treatment targets the pathological angiogenesis. Despite treatment,
many children suffer ongoing vision impairment. We are examining the major factors involved in the
development of ROP in order to develop more successful clinical treatments.
Projects available:
Project 1: Investigation of potential therapeutic targets aimed at inhibiting
photoreceptor death in a mouse model of retinal degeneration
Dr Kirstan Vessey and Dr Erica Fletcher
The excitatory neurotransmitter, extracellular ATP, acts via two classes of receptor, P2X and P2Y. The
P2X7 receptor (P2X7-R) is a unique member of the P2X receptor family of ligand gated ion channels. It
requires higher concentrations of ATP to become activated and when stimulated by ATP it not only
conducts cations, but following prolonged stimulation can conduct larger molecules, ultimately causing
cell death. As a result activation of the P2X7-R has been studied as a mediator of inflammation, cell
death and neural degeneration. This project will examine whether pharmacological blockade or knock
out of the P2X7-R slows photoreceptor loss and restores visual function in a mouse model of retinal
degeneration.
13
Project 2: The role of microglia in Age-Related Macular Degeneration
Dr Andrew Jobling and Dr Erica Fletcher
AMD is a major cause of vision loss in the older community. Recent work has indicated that
inappropriate activation of the immune response may play a role in the development of AMD. Retinal
microglia, the resident immune cells within the retina, are thought to play two alternative roles, one
being neuroprotective and the other resulting in neuronal cell death. Using a model system in which a
major signalling mechanism is “knocked out” in retinal microglia, we will investigate whether these
microglia are critical in both the protection of retinal neurons and what factors are altered that result in
retinal neuronal death. This project will involve the use of wide ranging techniques such as
immunohistochemistry, molecular biology and in vitro cell culture. Ultimately this study with detail why
microglia are critical to normal retinal function and what factors are involved in the development of
retinal degenerations such as AMD.
Project 3: The role of histamine in the development of the retina
Dr Una Greferath and Dr Erica Fletcher
Histamine is a biogenic amine involved in immune response and acts as a neurotransmitter in the brain.
We have very new and exciting evidences that histamine plays a role in the development and correct
lamination of the retina: mice, which cannot produce histamine (HDC-KO mice) have a disrupted
photoreceptor layer and significant photoreceptor loss in adulthood. We aim to investigate the
mechanism by which histamine controls photoreceptor development. We will need to localise histamine
and its receptors during development, evaluate the effect of histamine on photoreceptors in vitro and try
to rescue photoreceptor development by histamine supplementation. This project will involve the use of
techniques such as immunohistochemistry, molecular biology and in vitro cell culture.
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Autonomic and Sensory Neuroscience Laboratory
Principal Contacts
Assoc Prof James Brock ([email protected])
Professor John Furness ([email protected])
Dr Daniel Poole ([email protected])
Dr Tony Frugier ([email protected])
See Our Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/furness/index.html
Projects concern normal functions, animal models of disease, and studies of human tissues. Studies
focus on the autonomic nervous system, sensory innervation of organs, and the tissues innervated by
autonomic and visceral sensory neurons.
Technologies include: high resolution confocal microscopy; live cell imaging; molecular biology; in vivo
physiology; physiology of isolated organs; receptor signalling and trafficking; electrophysiology at the
single cell level; immunohistochemistry; in situ hybridisation histochemistry; quantitative PCR
For General Enquiries, contact Professor John B Furness: [email protected]
Digestive Health and the Enteric Nervous System: From Molecule to Human
Program Leader: Dr Daniel Poole, [email protected]
Altered function of the digestive system, as occurs following use of opiate analgesics (e.g. morphine) or
in irritable bowel syndrome and inflammatory bowel disease, can be debilitating and markedly reduce
quality of life. Our research focuses on identifying and characterising mechanisms of cellular signalling
in neurons of the intestine and examining dysregulation of this signalling under pathophysiological
conditions. Projects cover many aspects of G-protein coupled receptor signalling and trafficking, in
particular the roles of key molecules in the generation of opiate-induced bowel dysfunction and
inflammatory bowel disease. These projects have a high degree of translational relevance, and
encompass studies from the cellular and molecular level through to experiments on human tissues.
Transplantation science: Improving the survival of transplanted organs
Program Leader: Dr Tony Frugier, [email protected]
th
Transplantation of organs is a true miracle of the 20 century. Thousands of people are alive because
of their functioning donor kidneys, livers or other organs. However, decreased function of the donor
organs persists. One of the major problems is the challenge to tissues of the loss of oxygen and the
damage that can be caused by a surge of new oxygenated blood (this damage is reperfusion injury).
One of the greatest challenges is transplanting the intestine. In this project you will work alongside
transplant surgeons, research scientists and gastroenterologists to find solutions to the problems
precipitated by ischemia and reperfusion.
Nutrition: How nutrients modify digestive function
Program Leaders: Prof John Furness, Dr Brid Callaghan, Dr Daniel Poole. Contact Dr
Callaghan: [email protected]
It is becoming clear that nutrients strongly influence digestive behaviour. Some people are unable to
escape from a feeding and digestion cycle that leads to obesity. On the other hand, additives
(micronutrients) are added to feed to accelerate animal rates of maturation and to increase muscle
mass. Athletes also modify their diets to change body composition.
What do food additives actually do? We do not know. In this project you will work with a team of
researchers to identify receptors for micronutrients and to unravel their mechanisms of action.
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Spinal Cord Injury: Failure of bladder, bowel and blood pressure control
Program Leaders: Dr James Brock, Prof John Furness. Contact Dr Brock,
[email protected]
The trauma of Spinal Cord Injury is terrible, but living with the consequences is even worse. Injury to
the spinal cord disconnects the brain from autonomic centres in the spinal cord. Blood pressure control
is lost, and the person with SCI is subjected to large swings in blood pressure. Cardiovascular
problems are the largest cause of death in these patients. Also, bladder and bowel continence is lost,
and problems of bladder integrity, leading to often severe infection, emerge. This group of projects
involves clinicians, carers for the SCI injured and scientists determined to treat the consequences of
SCI.
Finding novel receptors important for breast and other cancers
Program Leaders: Prof John Furness, Dr James Brock, Dr Brid Callaghan. Contact Dr
Furness, [email protected]
One of the greatest challenges for therapeutics is the identification of target molecules. A related
challenge is to identify small therapeutic molecules that can be used in treatment. Proliferation of
cancer cells is reduced by the feeding hormone, ghrelin, acting at a so far unidentified receptor. This
project utilises pharmacological assays, molecular biology and medicinal chemistry to characterise the
receptor and to identify molecules that have potential in cancer treatment.
Mechanisms regulating neural control of arterial vessel constriction
Program leader: Dr James Brock, [email protected]
The brain controls blood pressure in part by regulating the activity of nerves that regulate the
constriction of small arteries and arterioles. Previous studies have investigated neural activation of
these vessels when they are isolated from the body, but to evoke constriction it is necessary to use
stimulation conditions that are considered unnatural. We hypothesize that this because the vessels are
removed from the normal influences of circulating hormones and substances produced locally within the
tissues that they supply with blood. This project uses pharmacological assays to investigate the actions
of these hormones and tissue-derived substances on nerve induced constriction of small arterial
vessels. A long-term objective is to study neural regulation of these vessels in situ.
Structure function relationships in peripheral sensory nerve terminals
Program Leaders: Dr James Brock, Dr Jason Ivanusic
[email protected]; [email protected]
Peripherally located small diameter sensory nerve terminals are critical for coding pain and changes in
temperature, but our current lack of understanding of how they work limits our ability to target specific
therapies associated with these sensory modalities.
This study uses a combination of
electrophysiological recording, neuroanatomical tracing, immunohistochemistry and high resolution
confocal microscopy to provide new insights into how sensory signals are transformed into action
potentials in sensory nerve terminals that code for pain and changes in temperature.
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Neuron Development and Plasticity Laboratory
Dr Jenny Gunnersen (Room E640, ph 8344 6065)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/gunnersen/index.html
How do neurons make connections in the developing brain?
The aim of our research is to understand how neurons become connected to
each other to form functional circuits. We investigate the formation of
dendrites (branches) and inter-neuronal connections (synapses) in
developing neurons in order to understand these processes in normal
development and disease. Many neurological disorders are characterized by
abnormal synaptic connectivity and changes in the number and strength of
synaptic connections occur during learning and memory formation (termed
plasticity). If we can understand how dendrites and synapses develop and
change in the healthy brain, this knowledge will help us decipher the aberrant
molecular pathways responsible for many cognitive disorders including
mental retardation, epilepsy and schizophrenia.
Honours and PhD projects are available to investigate these questions, focussing on the roles of key
molecules (e.g. Seizure-related gene 6 or Sez-6, Wnts, cytokines, Ndfip1) and their signalling pathways.
Sez-6 is a protein that is expressed in the developing
Sez-6 in neurons (green), internalized from
brain and in adult neurons in regions important for
the cell surface
learning and memory. To investigate the role of Sez-6,
we produced a knockout mouse in which the Sez-6
gene was inactivated. Analyses of neurons in the cortex
revealed that the dendrites of these neurons were
abnormal, that the neurons were less easily excited by
electrical stimulation and that there were fewer
synapses providing excitatory input to these neurons.
Abnormal Sez-6 function may be linked to the development of epileptic seizures and a Sez-6 family
member (Sez6L2) is a candidate gene for autism. Furthermore, the dendrite and synapse abnormalities
seen when Sez-6 is lacking are also seen in a number of mental retardation and neurodegenerative
conditions.
To study the complex molecular pathways regulating the development of neuronal branches and
synapses, we use a range of experimental approaches. Synapses in developing neurons are
fluorescently labelled using antibodies to pre- and post-synaptic markers. We are assessing protein
functions in the mature brain and in specific types of neurons using tissue-specific gene knockout or
gene knockdown approaches. We also assess molecular interactions and signalling pathways using
molecular biological and protein biochemical techniques.
Molecular mechanisms of maladaptive plasticity in neuropathic pain
Supervisors: Dr Jenny Gunnersen and Dr Maja Lovric, Department of Anatomy and
Neuroscience; A-Prof Christine Wright, Department of Pharmacology
Neuropathic pain is a type of chronic pain caused by nerve damage, either from traumatic injury or
diseases of the sensory nervous system. Pain may persist for months or years after the original injury,
responses to mild stimuli may be amplified and pain may occur in response to normally innocuous
stimuli. In experimental pain models, it has been possible to measure changes in the structure and
function (termed plasticity) of neurons that transmit sensory information to the brain. This “maladaptive
plasticity” is now recognized as an important pathological mechanism contributing to neuropathic pain.
Recent evidence indicates that Sez-6 knockout mice have altered susceptibility to developing
neuropathic pain. This project will investigate nociceptive neurons in Sez-6 knockout mice, using
immunohistochemistry and Golgi-Cox tracing, to correlate neuronal changes with pain development.
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Is Sez-6 important for learning and memory in the adult brain?
Supervisors: Dr Jenny Gunnersen and Dr Kathryn Munro, Department of Anatomy and
Neuroscience
When we examined neurons of Sez-6 mice that had developed from conception with no functional Sez6, fewer excitatory synapses were formed. As a consequence, the performance of these mice in
behavioural tests of cognition and motor function was affected. This project will investigate the
importance of Sez-6 in the mature brain using a conditional strategy to delete the Sez-6 gene in
neurons that are required for spatial memory. Conditional knockout mice will be tested in tests of
learning and memory.
Secreted signalling molecules in excitatory synapse development
Supervisors: Dr Jenny Gunnersen and Dr Kathryn Munro, Department of Anatomy and
Neuroscience, Dr Clare Parish, Florey Neuroscience Institutes
Proteins secreted from neurons act on nearby neurons to influence their growth and the formation of
synaptic connections. Several projects are available to examine the effects of different factors (including
secreted forms of Sez-6 and members of the Wnt and cytokine families) on the growth of neuronal
arbors (dendrites, axons) and synaptogenesis.
The role of TCF7L2 (TCF4) in regeneration after spinal cord injury
Supervisors: Dr Jenny Gunnersen, Department of Anatomy and Neuroscience, Dr Matthew
Digby, Dept. of Zoology
The transcription factor TCF7L2 (also called TCF4) is a downstream effector of the canonical Wnt
pathway that is important for cell fate/differentiation in development. TCF7L2 is downregulated in
response to spinal cord injury and this downregulation is associated with the failure of the mature spinal
cord to regenerate after injury (Mladinic M et al., 2010, Brain Res. 1363, 20-39). Interestingly, different
forms of this transcription factor have different, even opposite effects. This project will investigate the
different splice-isoforms of the messenger RNA for TCF7L2 in the mouse spinal cord in injury models
and determine the cell type that expresses TCF7L2 in the spinal cord.
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Stem Cell Genetics Laboratory
Identification of factors that regulate stem cell development in Drosophila
and mice
Dr Gary Hime (Room E637, ph 8344 5796)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/hime/index.html
Stem cells are the key to organ regeneration and tumour growth
Many differentiated but renewable cell types in vertebrates are derived from relatively small populations
of dedicated precursors, or stem cells. The ability to replenish differentiated cells depends on the
continued survival and proliferation of their respective stem cell populations. Stem cells are not only
important for regeneration of healthy tissues but also play a key role in pathogenesis. Recent studies
have demonstrated that all cells in solid tumours do not play equal roles but a small fraction of cells, the
so-called cancer stem cells, contribute to the unlimited growth of the tumour and re-occurrence after
tumour resection. If we are to realise the goals of re-programming tissue differentiation, growing organs
for transplantation in vitro, regeneration of damaged organs in vivo and targeted effective treatments for
cancer it is essential that we understand the molecules and mechanisms that stem cells utilise for
renewal and differentiation.
Drosophila and mouse organs – complimentary models of stem cell function
The identification of mechanisms that regulate asymmetric division, daughter cell mitotic amplification
and stem cell differentiation have been difficult to ascertain. These types of studies benefit greatly from
the analysis of simple, genetically tractable systems. For these reasons we have chosen to focus on
two stem cell niches in Drosophila (male germ line and intestinal) and one in the mouse (intestinal) as
models for stem cell systems.
A rosette of germline stem cells (expressing the Snail family zinc
finger protein Escargot, green) can be observed surrounding the
somatic stem cell niche in the Drosophila testis.
Projects are available within the following areas:
(1) Analysing the role of transcriptional repressors in Drosophila and mouse
stem cells
(In conjunction with Dr. Helen Abud, Monash University)
We have shown that members of the Snail family of transcriptional repressors are required in diverse
stem cell populations. This role has been conserved through evolution of animals as Snail family
members can be found in stem cells from Drosophila to mouse. This project involves using genetically
modified Drosophila or mouse strains combined with RT-PCR, microarrays and immunostudies to
identify what factors are being repressed by Snail family proteins in stem cells.
The mouse Snail family member, Snai1, is found in the nuclei of
intestinal stem cells that are found adjacent to the Paneth cells
(purple) as well as the nuclei of undifferentiated transit amplifying
cells.
From Horvay et al., (2010)
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(2) Analysing genetic roles of translational repressors in Drosophila stem
cells
(In conjunction with Dr. Nicole Siddall)
We have identified three RNA-binding proteins, Musashi, Real Musashi and Held-Out-Wings (HOW)
that are required to either prevent stem cells from differentiating in the stem cell niche or are required to
regulate the cell cycle of stem cells. This project utilises a variety of genetic, molecular biology and
immunostaining techniques to identify the roles of these proteins in different stem cell populations.
The ring of germline stem cells (red) can be observed
surrounding somatic hub cells (blue).
Loss of musashi (msi) function results in loss of stem
cells as they prematurely differentiate and lose
contact with the niche.
From Siddall et al. (2006)
(3) Analysing the role of a novel zinc finger protein in stem cell maintanence
(In conjunction with Dr. Greg Somers, LaTrobe University)
We have cloned the gene associated with a mutation that results in expansion of stem cells in the
Drosophila testis. The gene encodes a novel protein that contains 21 zinc finger domains. We do not
know what molecular processes are being regulated by this protein and the project will utilise molecular
genetics and immunostudies to determine its role in stem cell maintenance.
Expression of the stem cell marker
Escargot (green) is normally only
observed in the stem cell niche. In the
XP265 mutant we see stem cell markers
throughout the testis and disorganised
patterns of differentiation.
(4) How does regulation of nuclear importation affect stem cell maintenance
(In conjunction with Prof. K. Loveland, Monash University)
We have identified a nuclear importation factor that is required for maintenance of the germline stem
cell population. We will use immonostaining and confocal microscopy to determine the effects of
mutations of this factor on stem cell maintenance and conduct genetic interactions to identify proteins
that are imported by this factor into the stem cell nucleus.
References:
K Horvay, F Casagranda, A Gany, GR Hime and HE Abud (2011) Wnt signalling regulates Snai1
expression and cellular localisation in the mouse intestinal epithelial stem cell niche. Stem Cells and
Development, 20(4): 737-745
AC Monk, NA Siddall, T Volk, BA Fraser, LM Quinn, EA McLaughlin and GR Hime. (2010) The RNAbinding protein HOW is required for stem cell maintenance in the testis and for the onset of transit
amplifying divisions. Cell Stem Cell, 6: 348-360
NA Siddall, EA McLaughlin, NL Marriner and GR Hime. (2006) The RNA-binding protein Musashi is
required intrinsically to maintain stem cell identity. Proc. Natl. Acad. Sci. U.S.A. 103:8402-8407.
20
Neural Mechanisms of Bone Pain
Dr Jason Ivanusic (Room E724, ph 8344 7254)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/ivanusic/index.html
Pain associated with osteoarthritis, bone marrow oedema, bone cancer, fractures, osteoporosis and
osteomyelitis causes a major burden on individuals and health care systems in Australia. This burden is
expected to increase with advances in modern medicine that prolong life expectancy, because many of
the conditions that cause the pain are intractable and develop late in life. The pain is poorly managed by
current treatment strategies, and this likely results from our lack of understanding of the mechanisms
that generate and maintain bone pain.
The broad objective of the work done in this laboratory is to determine the fundamental neural
mechanisms that account for the perception of bone pain. This may lead to a better understanding of
ways to treat bone pain.
A number of projects are being conducted by the laboratory. The projects may be pursued as part of a
PhD, Masters, BMedSc or Honours degree, and any interested parties should contact Dr Jason
Ivanusic for further details.
Project 1: Changes in the neuro-chemical phenotype of primary afferent
neurons that innervate bone in an animal model of inflammatory bone pain.
It is generally assumed that the primary afferents neurons that innervate bone have a similar neurochemical phenotype to primary afferent neurons that innervate cutaneous and muscular tissues.
However, it is unclear whether the neurotransmitters and neuropeptides contained within boneassociated afferents are the same as in other fibre types, such as cutaneous, joint or muscle afferents.
The aim of this project is to investigate the neuro-chemical phenotype of primary afferent neurons that
innervate bone and to compare them to neurons that innervate skin, under normal conditions and in a
model of experimentally induced inflammatory bone pain. Small volumes of neuro-anatomical tracers
will be placed into a number of bony tissue locations (e.g. cortical, medullary and periosteal) or skin.
After an appropriate survival time and subsequent processing, the distribution of labelled neurons in the
peripheral sensory ganglia (DRG) will be determined using fluorescence microscopy, and the neurochemical phenotype of these neurons will be examined by using immuno-histochemical labelling of
common neurotransmitters, receptors and neuropeptides. Changes in the neuro-chemical phenotype of
these neurons will be examined in animals that have experimentally induced inflammatory bone pain,
because such changes may reflect mechanisms that are (at least in part) responsible for chronic or
persistent pain associated with inflammation. Students can expect to gain experience in animal
handling, behavioural testing, anaesthesia and surgery, dissection, histological and immunohistochemical processing, and fluorescence microscopy.
Project 2: Changes in neurotrophin signalling molecules in primary afferent
neurons that innervate bone in an animal model of inflammatory bone pain.
Neurotrophins have a well documented role in the developing nervous system, have recently also been
implicated in pain in the cutaneous and visceral systems, but are not well studied in models of bone
pain. The aim of project 2 is to determine whether sensory neurons that innervate bone have the
necessary receptors and proteins required for a role of neurotrophins in signalling bone pain, and to
further determine if there is a change in neurtrophin receptor or protein levels subsequent to
inflammation that could account for increased sensitivity to pain in an animal model of inflammatory
bone disease. The same experimental approach described for project 1 will be used. Students can
expect to gain experience in animal handling, behavioural testing, anaesthesia and surgery, dissection,
histological and immuno-histochemical processing, and fluorescence microscopy.
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Project 3: Investigation of the central nervous system areas associated with
bone pain.
Whilst Projects 1 and 2 will provide valuable data regarding the first part of the pathways that relay
sensory information from bone to the brain, it will not allow for investigation of the second and third
order neurons of these pathways because the neuro-anatomical tracers used are incapable of moving
across synapses. The aim of project 3 is to determine which parts of the CNS are involved in the relay
of information about pain from bone to the brain. This will be achieved by examining the distribution of
FOS expression in the CNS following stimulation of the rat tibia with noxious mechanical stimuli, and
under the condition of experimentally induce inflammatory bone pain. FOS is a protein marker for
neuronal activity and its distribution following activation of neuronal pathways can be examined
following a standard immuno-histochemical staining procedure. We have results which show increased
FOS expression in neurons of the dorsal horn of the spinal cord following noxious mechanical
stimulation of bone, have combined this approach with retrograde tracing techniques to determine the
target of these activated neurons, and plan to continue using these approaches to further define brain
areas associated with noxious stimulation of bone. Co-localization of markers for common
neurotransmitters and neuropeptides with the FOS protein may also be examined with the aim of
identifying putative local neuronal circuits involved in the transmission of information about bone pain.
Students can expect to gain experience in animal handling, behavioural testing, anaesthesia and
surgery, dissection, histological and immunohistochemical processing and fluorescence microscopy.
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Neural Development, Injury and Pain Laboratory
Laboratory Heads:
Professor Janet Keast (Room E720, phone 9035 9759)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/keast/index.html
Dr Peregrine Osborne (Room E641, phone 9035 9716)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/osborne/index.html
Postdoctoral fellows: Dr Adam Wallace, Dr Prajni Sadananda, Dr Priscila Cassaglia, Dr
Samantha Passey, Dr Sophie Payne.
We joined the Department in February 2012 and in our newly refurbished laboratories lead a multidisciplinary research team that investigates the neurobiological mechanisms underlying a number of
clinical conditions. We aim to understand the neurobiological causes of health problems such as
visceral pain, neuropathic pain (e.g., spinal cord injury pain), and conditions arising from developmental
disorders or disease of the urogenital system. We are especially interested in how the mechanisms that
control the development of neuronal circuits are re-activated in the adult to restore normal function or
cause dysfunction. The main focus of our developmental and neuronal regeneration studies is the
complex but fascinating neuronal circuitry of the pelvic ganglia and lumbosacral spinal cord.
Our research uses a variety of experimental approaches, including neuroanatomy, electrophysiology,
microsurgery, tract tracing, immunohistochemistry, confocal microscopy, neuronal cultures, cell and
molecular biology, live cell imaging and behavioural assays. Our work is supported by the NHMRC and
the US National Institutes of Health.
Project 1: Development of sacral autonomic circuits
Prof Janet Keast (supervisor), Dr Adam Wallace
The urogenital tract is controlled by autonomic neurons in the pelvic ganglia (also known as the pelvic
plexus or inferior hypogastric plexus in people). In comparison to other parts of the autonomic nervous
system, these ganglia are very unusual. For example, they are very different in males and females, and
they continue to be very sensitive to actions of steroids, even in adults. They are also mixed
sympathetic-parasympathetic ganglia, leading to questions of how these ganglia develop, and how their
connections with two different regions of the spinal cord (lumbar and sacral) are determined correctly.
Very little is understood about how this part of the nervous system develops, so there are a number of
possible projects in this area. One project could focus on how and when the male and female
differences in these neurons are determined. Another project could investigate the way in which spinal
preganglionic neurons correctly direct their axons to the appropriate group of neurons in the pelvic
ganglia. How and when sensory and motor connections are made with different urogenital tissues is
also of great interest. Techniques performed in this project include immunofluorescence, 3D-imaging
and reconstruction, tract-tracing in vitro and in vivo, microdissection of mouse embryos, use of reporter
mice.
Project 2: Improving the recovery of injured visceral nerves
Prof Janet Keast (supervisor), Dr Samantha Passey
Visceral sensory and motor (autonomic) nerves that control urogenital function are often damaged by
common surgeries such as prostatectomy, hysterectomy, bowel resection and other procedures
performed on related organs. As this damage is a cause of long-term or permanent visceral and sexual
dysfunction, we are studying how to promote functional recovery by stimulating and directing growth of
the injured neurons. To do this, we have established robust microsurgical models that can be used to
study relevant types of neuronal injury. These are being utilized for in vitro studies of neurotrophic and
steroid signalling pathways in isolated adult autonomic and sensory neurons maintained in culture;
anatomical tracing and confocal microscopy studies to analyse the recovery of connections after injury;
and in vivo studies to stimulate nerve growth using various physiological and pharmacological
strategies.
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Project 3: How nociceptor neurons change in a rodent model of
endometriosis.
Prof Janet Keast (supervisor), Dr Jane Girling (Department of Obstetrics and Gynaecology,
Royal Women’s Hospital)
Endometriosis affects 8-10% of reproductive-aged women. Women living with endometriosis endure
chronic pelvic pain, including severe menstrual pain, pain during sexual intercourse and pain during
defecation. The personal and healthcare costs of endometriosis are huge. How this pain develops is not
understood, althought various types of sensory and autonomic nerves are present in endometriotic
lesions and are thought to be involved. There are also distinct patterns of nerve fibres present in the
uterus that show an abnormal distribution in women with endometriosis. In this project we will use a
rodent model of this clinical condition to investigate how nociceptor neurons and their target neurons in
the spinal cord change during the development of endometriosis. Techniques relevant to this project
include: microsurgery, immunofluorescence and confocal microscopy, neuronal cultures, neuronal
growth and receptor internalisation assays.
Project 4: Neuroanatomical specialisation of pelvic sensory and pain
pathways
Prof Janet Keast, Dr Peregrine Osborne (supervisors)
Our understanding of peripheral sensory and pain processing is distorted by the overwhelming focus of
past studies on the afferent input to the lumbar enlargement. As a result we know comparatively little
about the organisation of sensory pathways that provide input to other spinal cord segments. This
project will focus on the sensory input to the sacral spinal cord. The sensory pathways at this spinal
level are critical to the normal functioning of visceral organs (including urogenital tract and bowel), sex
organs, and caudal somatic structures. Our group has contributed significantly to the progress made in
understanding the organisation of this important sensory system. In this project, you will use in situ
hybridisation techniques and apply them to a molecular pathway analysis of sensory neurons in the
pelvic dorsal root ganglia. Other techniques relevant to this project include: microsurgery,
immunofluorescence and confocal microscopy.
Project 5: Neuropharmacology of the GDNF family of neurotrophic factors in
peripheral nerve regeneration and pain.
Prof Janet Keast, Dr Peregrine Osborne (supervisors)
Nerve growth factor (NGF) is an endogenous neurotrophin that is released by tissues and targets
peripheral sensory neurons to cause peripheral sensitization and pain. Strategies targeting NGF
signalling are now being tested in clinical trials for the treatment of chronic pain. We, and others have
found that a majority of sensory neurons also express receptors targeted by another class of
neurotrophic factor, the GDNF-family ligands (GFLs). This has led to us to study the functions of GFL
signalling in normal and damaged peripheral sensory neurons. In this project you will contribute to
research performed in collaboration with Professor Mart Saarma (Institute of Biotechnology, University
of Helsinki). Applicable techniques include microsurgery, neuronal cell culture, microfluidics, nerve
growth assays, live cell imaging, and electrophysiology.
Project 6: Mechanisms of central pain after spinal cord injury (SCI)
Dr Peregrine Osborne, Prof Janet Keast (supervisors), Dr Priscila Cassaglia,
Dr Prajni Sadananda
Spinal cord damage caused by injury or disease is one of the most common causes of central
neuropathic pain. This type of pain can be experienced in the absence of any sensory stimulation and
cannot be successfully treated in around two thirds of patients. In this project you will study
neuroplasticity in the rat spinal cord that could contribute to increased pain sensitivity (hyperalgesia)
and spontaneous pain after SCI. Techniques to be used this project are immunohistochemistry, digital
and confocal microscopy incorporating image analysis, surgery and retrograde tracing, and behavioural
testing.
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Project 7: New uses of local anaesthetics in the treatment of visceral pain
Dr Peregrine Osborne, Prof Janet Keast (supervisors), Dr Prajni Sadananda
Mechanisms of visceral pain have been poorly studied due to the dominance of animal models of
somatic (skin and muscle) pain. This is unfortunate as visceral pain is one of the most common causes
of GP visits. In this project, you will contribute to our neuropharmacological research using rodent
models of bladder pain. We are studying transient receptor potential (TRP) channels and applying new
approaches for selectively inducing long-term anaesthesia in bladder pain sensing neurons
(nociceptors). Techniques to be used in this project are microsurgery, cystometry, live-cell imaging, and
electrophysiology.
Project 8: Application of multielectrode arrays (MEAs) in models of chronic
pain
Dr Peregrine Osborne, A/Prof Steven Petrou (supervisors)
Chronic pain is associated with hyperalgesia measured by increased sensitivity of the sensory system
to noxious and non-noxious stimulation. Hyperalgesia is caused by a sequence of events that include
peripheral sensitisation of sensory axons, central sensitisation of second order neurons in spinal cord,
as well as neuronal hyperexcitability and loss of inhibitory controls in the brain. Until recently, detecting
these changes in single neurons required laborious recordings of single units in peripheral axons, or
electrophysiological recordings from single cell with microelectrodes. In this project, you will develop
new assays for studying hyperalgesia in rodent models by making recordings with multielectrode arrays
(MEAs: see Fig. 4 Mouret et al., 2012, Nature Methods). These will be used to study different models of
hyperalgesia and to evaluate novel analgesic drugs.
Project 9: Sensory functions of the bladder urothelium in a co-culture system
Dr Peregrine Osborne, Prof Janet Keast (supervisors)
The urothelium lining the bladder is suggested to function as a sensory transducer that responds to
chemical and mechanical stimulation by releasing substances to activate sensory axons in the bladder
wall. The evidence supporting this hypothesis is compelling but is indirect. We have addressed this
shortcoming by developing a co-culture system in which sensory neurons and bladder urothelial cells
are maintained together. Using this approach we can now directly study intercellular communication
between urothelial cells and sensory neurons. As part of this project, you will refine this established
assay by using microfluidic chambers to establish compartmental cultures so that only the axons of
sensory neurons make contact with urothelial cells. Other techniques to be used in this project are
immunohistochemistry and live-cell imaging.
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Glia, Myelin Biology and Multiple Sclerosis Research
Principal Contacts
Prof Trevor Kilpatrick
Dr Ben Emery
CNS Myelination Laboratory
Dr Holly Cate
Myelin Repair Laboratory
Dr Simon Murray
Neurotrophin Signaling Laboratory
Dr Judith Field
Multiple Sclerosis Genetics Laboratory
See Our Web Pages: http://www.anatomy.unimelb.edu.au/researchlabs/murray/index.html
http://www.florey.edu.au/research/multiple-sclerosis
Our laboratories are located on the second floor of the new Kenneth Myer building.
Major Project Areas
How neurons react to the challenges of Inflammation and Ischemia – The roles of ion channels in
neurons – Visceral Pain – Spinal Cord Injury, seeking solutions to autonomic disturbances – The
actions of ghrelin in spinal cord autonomic centres.
We also run a modern histopathology laboratory, primarily for mouse work: http://www.apnhistopathology.unimelb.edu.au/.
There are opportunities for projects in our laboratories.
Overview of Research Laboratories
The multi-disciplinary strength of our research laboratories work collaboratively to address the complex
issues surrounding the normal development of myelin, how it responds following injury, potential
therapeutic strategies to promote repair, and the relevance this has to human disease. We combine
structural, physiological, pharmacological, neurochemical, genetic and in vivo studies of models of
central nervous system development and injury. We focus on animal models of disorders of the
nervous system, including irritable bowel syndrome, inflammatory bowel disease and the autonomic
consequences of spinal cord injury.
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CNS Myelination Laboratory
Dr. Ben Emery
Phone: 61-3-90356587
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/emery/index.html
General Overview:
The successful conduction of nerve impulses in the vertebrate nervous system is highly dependent on
myelin.
Within the central nervous system this myelin is produced by oligodendrocytes.
Oligodendrocytes are not only vital for the production of myelin and nerve conduction; they also have
important roles in the trophic and metabolic support of neurons.
Our primary interests include:
•
Understanding the molecular basis of neuronal-glial interactions
•
Elucidating how oligodendrocyte differentiation and myelination are regulated at the
transcriptional level
•
Identifying novel oligodendrocyte cell surface receptors and their role in myelination
Projects available:
We now have several projects investigating aspects of the transcriptional control of CNS myelination:
•
Use of biochemical, next-generation sequencing (ChIP-Seq and RNA-Seq) and cell culture
approaches to identify the gene targets of MRF during the myelination process
•
Use of conditional knockout and transgenic mice combined with experimental models of
demyelination to study the role of MRF in myelin repair and myelin maintenance in the adult CNS
•
Biochemical analysis of transcriptional complexes that drive the processes of oligodendrocyte
differentiation and CNS myelination to identify new players and clarify the functional relationships
between established pro-myelin transcription factors
Figure: Myelinating oligodendrocytes in the developing mouse optic nerve. The myelin sheath is stained
in red (myelin basic protein), the oligodendrocyte cell bodies in green (CC1 monoclonal)
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Myelin Repair Laboratory
Dr. Holly Cate
Phone: +61 3 83445264
Email: [email protected]
Website: http://www.cns.unimelb.edu.au/research/Myelin Repair/
General Overview:
Our lab is interested in enhancing nervous system regeneration following demyelinating disease.
During demyelination, there is death of oligodendrocytes, which are the myelin producing cells of the
central nervous system. This cell death leads to demyelination of axons, axonal degeneration and
progressive impairment of nerve cell function. Our lab uses cell culture assays and animal models of
demyelination to determine the cellular and molecular changes in neural stem cell and progenitor cell
populations in response to myelin damage. We then use this information to identify molecular targets
for enhancing nervous system regeneration.
Our work has identified genes that are elevated in the brain during myelin damage. We are currently
investigating the effects of modulating of Bone Morphogenic Protein (BMP) signaling in mouse models
of demyelination. An understanding of how BMP and other endogenous factors modulate remyelination
and ways to promote their efficiency for regeneration is a step toward identification of potential
therapeutics.
Projects available:
We have the projects available in the following areas:
1.
The role of Bone Morphogenic Protein signaling in regulating neural stem cell differentiation
during demyelination and myelin repair
2.
Regulation of Oligodendrocyte progenitor cell differentiation during myelin injury
3.
Analysis of the effects of blocking BMP signalling in acute and chronic models of demyelination.
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Neurotrophin Signaling Laboratory
Dr. Simon Murray
Phone: 8344 5813
Email: [email protected]
Dr. Junhua Xiao
Phone: 8344 7572
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/murray/index.html
General Overview:
Demyelinating diseases of both the peripheral and central nervous system have a devastating human
impact. There is currently an incomplete understanding of the factors that initiate, promote and
maintain the interactions between neurons and glial cells that are vital for myelination. Our laboratory is
interested in increasing our understanding of some of the factors that regulate myelination and dissect
their primary mechanism of action. Our focus centres on a family of growth factors known as the
neurotrophins, and the influence they exert on both peripheral and central nervous system myelination.
We use a variety of molecular, cellular, biochemical and genetic techniques to investigate these events.
Projects available:
The influence of BDNF on CNS & PNS Myelination
We have recently identified that BDNF plays an important role in promoting both central and peripheral
nervous system myelination. Our data indicate that BDNF activates distinct receptors to regulate
myelination: it activates the receptor tyrosine kinase TrkB to promote CNS myelination, but activates the
neurotrophin receptor p75NTR to promote PNS cell myelination.
We are currently investigating the signalling events that mediate these distinct promyelinating signals,
and also developing low molecular weight novel peptides designed to selectively mimic the TrkB and
p75NTR agonist properties of BDNF. We have a number of projects that will undertake:
(i)
structural-based NMR studies to fully characterise the structure of the BDNF mimetic peptides,
and investigate the key residues involved in binding to their respective receptors
(ii)
biochemical assays to investigate the capacity of the BDNF mimetic peptides to activate their
respective receptors and initiate key intracellular signalling cascades
(iii)
in vitro myelination assays to investigate whether the the BDNF mimetic peptides can, just like
BDNF, promote myelination by oligodendrocytes and Schwann cells
(iv)
examine the signalling cascades and transcription factors activated by BDNF that promote both
central and peripheral nervous system myelination
These projects involve NMR spectroscopy, structural biology, routine cell culture as well as the
generation and co-culture of primary neurons, oligodendrocytes and Schwann cells. Through analysis
of these cultures by immunocytochemistry, Western blotting and other biochemical assays, we will
identify whether our BDNF mimetics can promote nervous system myelination and their mechanism of
action.
These projects will be undertaken in collaboration with investigators in the Department of Pharmacology
and at Bio21.
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Multiple Sclerosis Genetics Laboratory
Dr Judith Field
Phone: (03) 9035-6518
Email: [email protected]
Web page: http://www.florey.edu.au/research/multiple-sclerosis
General Overview:
Multiple sclerosis (MS) is a complex disorder of which the cause has yet to be determined. It is clear
however that both genetics and the environment play a role in the development of disease. Our work
aims to identify the genetic determinants of susceptibility to MS, and has resulted in significant
contribution to our knowledge of the genes that are involved. Many of the MS risk genes identified play
a role in the immune system (e.g. CD40 and IL2RA), and using a variety of both genetic and
immunological techniques, we also aim to determine the functional consequences of these genetic
associations in the development of MS.
Projects available:
1) Regulation of Interleukin 2 receptor alpha (IL2RA) DNA methylation in MS
MS is a degenerative disease of the central nervous system (CNS) in which oligodendrocytes, the
myelin-producing cells of the brain, are damaged and myelin is lost (demyelination). Both genetic and
environmental factors have been shown to contribute to the risk of developing MS. Interleukin-2
receptor alpha (IL2RA), also known as CD25, has been identified as a susceptibility gene in MS, and is
expressed by cells of the immune system, particularly regulatory T cells and CD4+ T cells in response
to activation stimuli.
Using experimental approaches which look at the methylation state of DNA at sites known as CpG
dinucleotides, we have found that there are differences between the methylation profile of the IL2RA
gene in DNA isolated from neural tissue samples compared to cells isolated from the peripheral blood of
humans. In particular 4 CpG methylation sites within the promoter region of IL2RA show different
methylation profiles, which correlate with differences in expression levels of IL2RA in neural tissue and
peripheral blood cells. This project will aim to investigate the effects of differential methylation at these
sites on gene expression, as well as determine whether the DNA methylation profile in cells expressing
IL2RA differs between individuals who have MS compared to those who do not.
2) Functional consequences of Multiple Sclerosis risk genes
Many of the genes that affect whether an individual is at higher risk of developing MS have been
identified. The involvement of the gene known as CD40 in the risk of MS was first discovered in an
Australian study. This gene is involved in the activation of cells of the immune system, and has also
been shown to play a role in the development of other autoimmune diseases. What we do not know is
how the genetic change in CD40 alters the normal function of the immune system, and therefore how is
leads to the development of MS in some people. In this project we aim to establish how the genetic
change in CD40 affects the function of the CD40 gene in MS, and how we can use this information to
develop therapeutic strategies for patients with MS. This project will focus on assessment of the CD40
protein levels in individuals carrying the MS-risk associated allele compared to those carrying the
protective allele, and the effect on stimulation dependent responses to gain a better understanding of
the functional consequences of altered expression in the development of MS.
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Early Brain Development Laboratory
Dr Peter Kitchener (Room E722, ph 8344 6746)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/kitchener/index.html
Project 1: Microglial invasion and migration into the embryonic brain
This project will attempt to elucidate the molecular mechanisms by which microglial precursors enter the
embryonic central nervous system (CNS). This will involve 1) a spatiotemporal description i.e. when and
where entry occurs; 2) a microscopic description e.g. what cell types are involved and their behaviour
over the course of the process; 3) delineation of any temporal or other constraints on the events; and 4)
investigating any candidate molecules that mediate the various aspects of the process. The primary
methods used will be histology (including live cell tracking) and immunochemistry.
Figure 1. Three ramified microglial cells (and three
capillaries) in the cerebral cortex of an Embryonic day
15 fetal mouse. These microglia are blood-derived
cells that invade the central nervous system and take
up permanent residence as it develops
Project 2: The formation of the cardiovascular control centres in brainstem
Dr Peter Kitchener and Assoc. Prof. Andrew Allen (Dept. Physiology)
The aim of this project is describe the development of the nuclei responsible for the regulation of vital
functions, especially cardio-vascular regulation. We are interested in the emerging view that problems in
cardiovascular regulation may have their origins in the early development of, and early environmental
influences on, these brain regions. We wish to determine the periods of the key events: the birth, growth
and differentiation of these neurons. With this knowledge, there can be a rational approach to
manipulating the plasticity of these control systems, providing a better understanding cardiovascular
regulation and also a promising avenue for understanding the pathogenesis of disorders such as
essential hypertension.
Figure 2. Tyrosine hydroxylase immuno-staining of neurons
in the locus ceruleus (LC) and the cardiovascular-regulatory
nucleus in the rostral ventral lateral medulla (RVLM) in an
Embryonic day 18 mouse brainstem.
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Behavioural Neuroscience at the Florey
Division Head: Professor Andrew J. Lawrence
Contact details: (03) 90356692
Email: [email protected]
Web Page: http://www.florey.edu.au/about-florey/our-people/staff-directory/208/andrew-lawrence
Addiction Neuroscience Lab Overview:
Our laboratory is investigating the neural pathways implicated in drug-seeking behaviour and
additionally, the association between mood disorders, stress and drug-seeking behaviour. For example,
alcohol is one of the most widely used and abused drugs in society, with an immense social, medical
and financial impact. The neuropharmacological basis of alcohol reward is an ongoing research area in
this laboratory. A major focus of our research has been the development of relevant rodent models of
addiction. We employ a number of genetic animal models, including inbred strains of rats and knockout
/ transgenic mice. Drug self-administration and behavioural parameters are measured to assess the
influence of specific receptors and emotional states such as anxiety and depression on drug-seeking
behaviour. Stressors can be used to examine the relationship between psychological state and relapse
to drug-seeking behaviour following a period of abstinence. These behavioural approaches are
complemented with a range of neurochemical, anatomical and molecular strategies to provide a
multidisciplinary strategy.
In addition, we are employing genetic approaches to investigate the neural substrates of drug-seeking,
drug-induced plasticity and relapse. This latter aspect is of critical importance, as the defining feature of
addictions is the chronic and relapsing nature of the disorder. Many of these research directions have a
common underlying theme of neural plasticity as a key mediator of altered behaviour patterns. To
facilitate these studies we have perfected the technique of intravenous self-administration of drugs of
abuse in rodents. Projects currently underway involve self-administration of opiates, cocaine and
nicotine. Together the use of different rodent strains, along with pertinent paradigms has enabled us to
address key questions pertinent to addiction, such as the identification of factors implicated in
behavioural responses to drugs of abuse and also the chemistry of relapse.
Project 1: The effects of toluene exposure on the adolescent brain
Supervisors: Dr. Jhodie R. Duncan and Professor Andrew J. Lawrence
Phone: 90357046
Email: [email protected]
Inhalant abuse (also known as “chroming”) is a major socio-economic problem in Australia, and a
rapidly growing drug of choice to reach a euphoric state. Inhalants such as paint, glue, hair-spray or
petrol are among the cheapest drugs in the community, and unlike illicit drugs, there are no legal
restrictions on their purchase, supply, possession or use. The typical onset of experimentation with
inhalants occurs earlier than with most other drugs of abuse, in the preteen years. Indeed the incidence
of inhalant abuse is greatest in the adolescent population with numbers as high as 26% of adolescents
abusing solvents; with 12-13 year old comprising nearly 50% of this population. For concern,
adolescence encompasses an extensive period of neuronal maturation in the brain (i.e myelination and
synaptic pruning) especially in regions associated with core executive and self-regulatory skills such as
inhibitory control and affect-regulation. Therefore abuse of inhalants during this time coincides with the
maturation of crucial cognitive and emotional brain structures. Furthermore experimental inhalant use
during adolescents is a significant risk factor for addictive behaviors later in life.
The aim of this project is to utilise a rodent model of adolescent toluene exposure via inhalation in order
to increase our understanding of the resultant neuroadaptations that may result in altered behavior later
in life. The project will investigate the effects of both acute and chronic adolescent toluene exposure on
longer-term brain function including the assessment of behavioural flexibility following prior exposure to
toluene during adolescence. This will be investigated using behavioural paradigms to test for factors
such as impulsivity, social interaction, perseverance and decision making. Results from these
behavioural studies will then be correlated to data on the neurochemical adaptations at the gene and
protein level in order to increase our understanding of the mechanisms that may be underlying altered
behaviour.
32
Thus results from this study will increase our understanding of the long-term effects of inhalant abuse
on resultant behavior therefore increasing our understanding of the pathways affected following toluene
exposure, especially if abuse occurs during adolescence when the yet mature brain may be more
susceptible to drug induced neuroadaptations.
Project 2: Identification of hypothalamic peptides that regulate rewardseeking
Supervisor: Professor Andrew J. Lawrence
Phone: 90356692
Email: [email protected] or [email protected]
In 2006 we identified the hypothalamic orexin neurons as a key component of the circuit involved in
alcohol self-administration and relapse to alcohol-seeking in abstinent rats. These neurons, while
located in the hypothalamus innervate the entire neuraxis. More recently we have demonstrated that
orexin receptors in the prefrontal cortex are implicated in cue-induced reinstatement (relapse) to
alcohol-seeking suggesting that ascending orexin pathways are involved in attribution of cue-salience
and behavioural reactions to conditioned cues.
While we have shown that hypothalamic orexin neurons are activated during relapse to both drug
(alcohol, opiate) and natural (sugar) rewards, there are also other hypothalamic non-orexinergic
neurons implicated in relapse. Recent ongoing studies in our laboratory suggest that a population of
hypothalamic neurons that express the receptor for leptin (LepRb) are activated during cue-induced
drug-seeking. These neurons also co-express the peptide neurotensin.
The current project will essentially have two components, (i) studies using a transgenic LepRb-YFP
mouse to map the LepRb-positive neurons activated during relapse and (ii) microinjections of drugs that
act on either neurotensin or leptin receptors directly into defined brain nuclei to determine the role of
these peptides in relapse.
Project 3: Neural circuitry underlying extinction of fear across development
Supervisor: Dr Jee Hyun Kim
Phone: 90356623
Email: [email protected]
Most anxiety disorders emerge during childhood, and individuals with childhood onset express more
severe symptoms than do individuals who have adult onset. In fact, there is growing recognition that
mental disorders may actually be developmental brain disorders and, as such, treatment strategies
should focus on the young population.
Currently, the effective treatments for anxiety disorders are cognitive-behavioural therapies that rely on
the process of extinction. Extinction is the decrease in fear responses expressed to a fearful stimulus
due to the repeated exposure to the stimulus without any aversive outcome. We have accumulated
powerful evidence supporting that extinction is erasure in juvenile rats whereas extinction is new
learning in adult rats. This developmental transition from erasure to new learning appears to be driven
by changes in the functionality and the circuitry between the amygdala, the hippocampus, and the
medial prefrontal cortex (mPFC). This project will characterise the functional organisation of that neural
circuitry using retrograde tracers and intracranial microinfusions using Pavlovian fear conditioning as a
model of post-traumatic stress disorder in developing rats.
33
Project 4: Extinction of drug-seeking: pathways & mechanisms
Supervisors: Dr Christina Perry, Dr Jee Hyun Kim and Professor Andrew J. Lawrence
Phone: 90357527
Email: [email protected]
Addiction is characterised by high rates of relapse with relapse episodes occurring even after years of
abstinence. Much of the research into addiction and relapse using animal models has focused on the
neurochemistry and neurobiology of reinstatement. However, understanding the mechanism of
extinction is crucial for developing more effective therapies and for preventing relapse. This project will
examine the neurochemistry underlying extinction of cocaine self-administration in rodents. It will focus
in particular on the role of the µ opioid receptor, and the NMDA receptor. Both of these are crucial for
mediating new learning that occurs during extinction. Rats will be trained to self-administer cocaine, and
then this response will be extinguished. A µ-opioid receptor agonist (morphine) or an NMDA receptor
antagonist (ifenprodil) will be applied during extinction sessions. The effect on responding during
subsequent extinction sessions, and during reinstatement produced in response to a cocaine prime will
be examined. We expect that these manipulations will delay extinction and increase reinstatement. A
better understanding of the neurochemistry of extinction would aid in the development of more effective
pharmaceutical adjuncts to behavioural therapies aimed at promoting abstinence in persons addicted to
cocaine and other drugs of abuse.
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Brain and Behaviour Laboratory
BEHAVIOURAL NEUROSCIENCE
Dr Mark Murphy (Room E616, ph 8344 5785)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/murphy/index.html
How do we think, feel, learn, remember and behave? We are beginning to investigate these questions
on a number of different fronts. The research we are involved in is searching for fundamental
mechanisms involved in learning and memory, emotions and emotional memory, and the basis of
individual variation in peoples’ behaviour.
Project 1: What circuits in the brain are involved in a particular behaviour?
The key way in which the brain works is as a complicated set of circuits, which link up various bits of
information, combine them, process them and finally set the body to act. Some of the primary circuits in
the brain, which are involved in a particular function, such as those involved in regulating movement,
have been reasonably well characterized. However, for the brain functions, which underlie higher order
behaviours, we have little idea of the brain circuits, which are involved. One of the reasons we know so
little is because there have been no good ways of defining these circuits. We have generated
transgenic animals, called fos-tau-lacZ mice, which will hopefully allow us to trace the circuits, which are
used for a brain function. These mice have been designed to express a marker protein in the axons and
dendrites of neurons, which have been activated by some functional stimulus. Thus, when the animal is
doing something, the neurons, which are involved in that function will “light up”, and so will their axonal
projections throughout the brain.
Our current projects with these mice involve looking for circuits involved with learning and memory,
processing of vision, and the processing of novelty.
Project 2: Plasticity in learning and memory.
What we hope to learn in our experiments above is which circuits are involved in the learning of a
particular event. But to understand the learning process fully, we need to understand what changes
occur in learning which result in the storage of that information in memory. Learning is thought to
involve alterations in existing brain circuits. The key is the synapse. This is the connection between
axon and dendrite, and it must be here that any changes in circuitry within the brain must be
established. So far, there have been no direct demonstrations of synaptic change underlying a learning
process. We are currently utilizing immunosorbent assays and confocal microscopy techniques to
examine changes in the expression of synaptic proteins, in mice, following training on learning
paradigms. We are using similar techniques to study the changes, which occur in the brain following
exposures to novel environments. These studies are interesting because novel environments have
powerful behavioural effects on animals, such as decreasing stress levels and improving their abilities in
complex behavioural tasks.
Project 3: Genetics of Behaviour.
The final area of research involves inherent components of behaviour. There now seems no doubt that
variation in types of behaviour, such as personality type, has a genetic contribution. Depending on the
type of behaviour, the genetic component contributes from 40% to 70% of the variation. One such
behavioural trait is associated with the way we cope with stressful events in our life. Our stress
response is a natural and important part of our behaviour and contributes to our survival and well-being.
However, a prolonged and heightened stress response can have very serious consequences such as
long term anxiety states and panic attacks, depression, and susceptibility to cardiovascular disease and
immune dysfunction. The way in which individuals respond to stress varies considerably and this
variation has a genetic contribution of at about 50%. Clearly, it would be useful to understand which
genes are involved in stress response, not only to gain a greater understanding of stress as behaviour,
but also to understand how to modify our responses to stress. We are involved in a study, which will
map the genes involved in stress response in an experimental animal model. The final aim of this study
is to identify the genes which are involved in the stress response, and how presumed variations in these
genes lead to different stress responses and different susceptibilities to the pathological conditions
associated with stress.
35
Project 4: Methods to ameliorate cognitive decline and memory deficits in
Alzheimer’s disease.
This project is being conducted in collaboration with Assoc Prof Graham Barrett, Department of
Pathology, University of Melbourne.
Alzheimer’s disease (AD) is an extremely prevalent cause of dementia. One of the major consequences
of AD is memory loss. It is characterized by death and degeneration of neurons, extracellular deposits
of amyloid plaque, and intracellular fibrillary tangles. Cholinergic neurons in the basal forebrain
comprise one of the earliest and most severely affected cell types in AD. Neurodegeneration of these
neurons gives rise to a severe cholinergic deficit, which is believed to be instrumental in the impairment
of memory and attention in AD. In the mature brain, the same cholinergic neurons are the only neurons
that express a receptor for the neurotrophin growth factors, p75. In the cholinergic system, p75 signals
the negative aspects of trophic regulation and thus acts as an inhibitor of cholinergic neuronal size,
growth and activity. It can also cause apoptosis, particularly in response to injurious stimuli. We have
previously shown that ablation of this p75 receptor has a remarkable effect on the cholinergic system in
normal mice. Ablation of p75 results in cholinergic neuron growth, increases cholinergic innervation of
the hippocampus, enhances hippocampal synaptic plasticity and improves spatial memory
performance. Our hypothesis is that reduction and/or ablation of p75 in a mouse model of AD will
alleviate the cognitive symptoms, primarily by its actions on the cholinergic system.
The project asks two closely-related questions; firstly, what is the role of p75 in the cholinergic
neurodegeneration that occurs in AD? Secondly, does stimulation of the cholinergic system by
removing or downregulating p75 produce an improvement in cognitive performance in AD? We have, in
fact, previously shown that removing p75 in normal mice enhances the cholinergic system and memory
performance in normal mice.
To help address these questions, we will conduct behavioural assays, including tests of spatial memory,
in Alzheimer’s mice with varying degrees of p75 downregulation. After these tests, the brains will be
examined histologically and biochemically. We will assess the changes in the size, number and activity
of cholinergic neurons. We will look for changes in cell death and amyloid deposition.
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Neurotrophin Signaling Laboratory
Dr. Simon Murray
Ph: 8344 5813
Email: [email protected]
Dr. Junhua Xiao
Ph: 8344 7572
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/murray/index.html
General Overview:
The focus of our laboratory centres on a family of growth factors known as the neurotrophins. The
neurotrophins play key roles in promoting nervous system development and health, exerting significant
control over the pathways that trigger neuronal life and death. Thus, the neurotrophins hold great
promise as potential therapeutic candidates. Towards this end, we are interested in understanding the
neurotrophins and how they signal, and have projects that examine the potential therapeutic benefit of
the neurotrophins. We use a variety of molecular, cellular, biochemical and genetic techniques to
investigate these events.
The laboratory is located on the second floor of the Melbourne Brain Centre, housed in the new
Kenneth Myer Building.
Projects available:
Project 1: Analysis of BDNF and its mimetics
The neurotrophin brain-derived neurotrophic factor (BDNF) regulates
multiple functions within the nervous system, in part due to the fact that
it signals through two distinct receptors: TrkB and p75NTR. A means of
selectively activating these receptors would clearly be desirable if one
is to unravel the effects of BDNF and exploit the effects mediated by
these distinct receptors therapeutically. We have designed and
synthesised small functional mimetics of BDNF, and recently
determined that these peptides are biologically active.
The small peptide mimetics of BDNF:
1.
peptide 6, a putative activator of TrkB
2.
cyclo-dPAKKR, a putative activator of p75NTR
We have projects investigating the properties of these novel peptides:
(i)
structural-based NMR studies to fully characterise the structure of the mimetics and investigate
their binding to their respective receptors.
(ii)
biochemical assays to investigate the capacity of the mimetics to activate their respective
receptors and initiate key intracellular signaling cascades
(iii)
in vitro assays to investigate whether the mimetics can, just like BDNF, promote both peripheral
and central nervous system myelination.
(iv)
in vivo assays to investigate whether the mimetics can promote remyelination following injury to
the peripheral and central nervous system.
This project will be undertaken in collaboration with investigators in the Department of Pharmacology
and at Bio21.
37
Project 2: Mechanisms of growth factor signaling
Nervous system development requires highly specific and co-ordinated growth factor signaling. Precise
control of signaling is dependent on a number of factors, such as presentation of the ligand, receptor
structure and activation, the expression of co-receptors and adapter molecules, and the negative control
or inhibition of signaling. One focus of our laboratory is to investigate neurotrophin signaling and how it
is positively and negatively regulated. In particular, we focus on the neurotrophins NGF and BDNF.
We are currently investigating aspects of neurotrophin signaling, including the form in which the
neurotrophins are presented and a number of novel molecules that interact with neurotrophin receptors.
We have several projects investigating how neurotrophin signaling is regulated by these molecules
which include:
(i) The precursor form of the neurotrophins, and their impact on cell signaling.
(ii) The Spred and Sprouty proteins, inhibitors of growth factor signaling.
(iii) The Neurotrophin Receptor Homolog, which modulates neurotrophin signaling.
(iv) The Sorting Nexin (SNX) proteins, which regulate the cellular location of receptors.
Using in vitro approaches, the successful applicant will learn a variety of skills such as tissue culture
techniques, cellular transfection, immunocytochemistry, SDS-PAGE and Western blotting,
immunoprecipitation, PCR and cDNA mutagenesis.
These projects will identify some of the vitally important cellular events that regulate neurotrophin
signaling and control the development and growth of the mammalian nervous system. Analyses such as
these will provide new insights and develop into approaches that may be utilised to halt the progression
of degenerative neurological diseases.
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Animal Brain Imaging using MRI
Professor Roger Ordidge
Phone: 8344 1953
Email: [email protected]
Project 1: Optimisation of MRI tractography by Diffusion-weighted MRI
This project aims to improve the accuracy and reliability of tracking nerve fibres in normal and abnormal
animal brain using MRI. Novel methodology will be developed in combination with unique equipment in
order to establish connectivity changes between different brain regions and the effects of injury and
disease.
Figure 1 shows: on the right, a normal image of a mouse brain: in the middle, an image of fibre track
density: on the left, an image representing fibre tracks in the brain section.
All images were obtained on a 4.7 Tesla animal MRI system in the Florey Neurosciences Institute using
a cryo-cooled receive coil.
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The Ion Channels and Disease Laboratory
Steven Petrou ([email protected])
Chris Reid
Our laboratory is located on the first floor in the Melbourne Brain Centre, Kenneth Myer Building, and is
fully equipped with state of the art neurophysiological and imaging capabilities. We are a 20 person
multidisciplinary team working on individual and joint projects in the neurosciences. Our primary interest
is in diseases and therapies that involve ion channels with a particular focus on epilepsy. In epilepsy
our work begins with clinical and genetics collaborators who identify gene mutations. Many of these are
in ion channels and we seek to understand how these mutated genes lead to behavioural seizures. x
We use a range of methods, appropriate to the scale of investigation and combine, genetic, molecular,
biophysical, computational, neurophysiological and behavioural approaches. In addition, our laboratory
houses the Australian Optogenetics Repository and we are well positioned to exploit this exciting new
method. The projects below give a sample of the work being undertaken and available for suitable
candidates.
Projects in network analysis of genetic epilepsy
Epilepsy impacts around 3% of the population and in many cases has clear genetic underpinnings. Our
laboratory has created several genetically engineered models of epilepsy that have helped provide the
most detailed understanding of how a single gene mutation can lead to behavioural seizures. Perhaps
the largest gap in our understanding lies at the level of the network that bridges cellular and synaptic
function with the actual seizure phenotype itself.
Project 1: Multi site patch clamp recording of cortical micro networks
Steven Petrou and Christopher Reid
In this project the candidate will be trained in the use of an emerging method in brain slice
electrophysiology that allows for the simultaneous intracellular recording of 4 connected neurons. Using
this recording mode it is possible to examine how neurons function in coupled micro networks in
epileptic and normal brains to lead to a deeper understanding of the functional basis of epilepsy. If the
candidate makes sufficient progress and is motivated this project may also expand into network
analysis using multiphoton imaging where 50 or more neurons in a living brain can be labelled with a
Ca2+ indicator dye and imaged in real time.
Project 2: High density multi-electrode array recording of in vitro networks in
epilepsy
Steven Petrou and Christopher Reid
In this project the candidate will use high density extracellular multielectrode array recordings to
investigate large scale network function. This a level of organization beyond that studied in Project 1
and will reveal fundamental properties of how the hippocampal and thalamocortical networks are altered
in genetic models of epilepsy. The goal of these studies is to not only understand more about the
neurobiology of epilepsy but also to create novel disease state models for creating anti-epileptic drugs.
The method will involve cutting fresh brain slices and using 60 site multi-electrode arrays that enable
electrical stimulation and recording from all sites simultaneously. Slices will be subject to various
stimulation and pharmacological protocols to reveal aspects about excitability, synaptic transmission
and plasticity.
Project 3: In vivo electrophysiological analysis in mouse models of genetic
epilepsy
Steven Petrou and Antonio Paolini
In this project the candidate will use multi-site in vivo unit recording in mouse models of genetic epilepsy
to investigate network function and dysfunction in freely moving mice. Using digital high density
electrode recording the candidate will implant multiple sites and then record from mice housed in a
controlled environment with video monitoring. One possible addition to these experiments is the
incorporation of optogenetic stimulation whilst recording to probe network function in connected
networks of behaving mice. This will provide some of the first views into how real time intervention of
networks modulates seizure initiation and termination.
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Project 4: The glass brain: "Connectomics" in epilepsy
Steven Petrou, Verena Wimmer and Kay Richards
Recent improvements in the histochemical method of optically clearing whole tissues and the joint
development of special optics that can image deep into them have created unprecedented views into
the wiring of networks. Changes in wiring of cortical neurons have been implicated in a number of
disorders such as epilepsy, schizophrenia and depression. In this project the candidate will prepare
brains from mice with fluorescently labelled neurons and use 2-photon excitation to create 3D images in
regions of the mouse cortex. By comparing normal and epilepsy models this work will begin to unravel
the changes that occur prior to and after the occurrence of seizures. This will shed important light on
the scale on which structural changes occur in epilepsy and will guide future experimental and clinical
work.
Project 5: MRI tractography in mouse models of genetic epilepsy: Creation of
prognostic and diagnostic structural biomarkers.
Steven Petrou, Kay Richards, Chris Reid, Alan Connelly, Donald Tournier and Fernando
Calamente
Our earlier classical histological analyses have shown that neuronal numbers and positioning are both
altered in genetic forms of epilepsy prior to the appearance of overt seizures suggesting that structural
changes precede epilepsy. These changes, however, would be below the level of detection of current
clinical MRI scanning technology and have led to the potentially erroneous conclusion that idiopathic
generalised epilepsy (IGE) is characterised by a complete absence of structural change. By combining
recent developments in super resolution MRI (developed by members of the supervisory team) and high
field MRI acquisition (16.4T) the candidate will seek to reveal structural changes, or biomarkers, that
precede or are a consequence of epilepsy. Because these approaches are directly translatable into the
clinic any finding could be rapidly tested in patients. The candidate will develop skills in preparing fixed
mouse brains for MRI scanning at 16.4T at the Queensland Brain Institute for analysis using the MRtrix
suite of software on a custom workstation to compare brains from control and genetic mouse models.
Project 6: High content automated analysis of ion channels in epilepsy
Steven Petrou, Carol Milligan and Chris Reid
Discovery of gene mutations in neurological disorders such as epilepsy is outstripping the ability to
functionally validate them. Because many epilepsy genes code for ion channels we have established
high content automated patch clamp platforms based on the Nanion Patchliner 16 and the Fluxion HT
64 systems to bridge the "discovery" gap between genetics and functional validation. Several new
mutations have been found by our geneticist collaborators that are awaiting detailed functional analysis
and the candidate will first have to produce mutant cDNAs then transiently transfect into HEK293 or
CHO cells prior to analysis on the automated platforms. Candidates will be trained in the necessary
molecular biological methods and then in ion channel electrophysiology and will work closely with a
senior member of the team to ensure success.
Project 7: Optogenetic modulation of the area tempestas - an epilepsy hot
spot.
Steven Petrou, Antonio Paolini, Chris Reid
Several lines of study have recently converged to reveal a new target for controlling epileptic seizures.
Early work by Piredda and Gale (Nature 1985, 317:623) provided unequivocal evidence that the
prepiriform cortex, subsequently coined the “area tempestas”, was a hot spot for initiation and spread of
epileptic seizures. Within this region a population of specialised inhibitory neurons called neurogliaform
cells (NG) shows a stereotypic pattern of firing that implicates them seizures. In this project the
candidate will use in vivo electrophysiological recording and optogenetic stimulation to examine real
time modulation of the control of seizures to develop a role for the in vivo function of NG cells and
explore their potential utility in seizure suppression.
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Project 8: Exploring the role of GABA mediated tonic inhibition in depression
Steven Petrou, Robert Richardson and Chris Reid
Depression is a serious neurological disorder that can impact people of all ages and genders. Elegant
studies led by Istvan Mody at UCLA have shown that in post-partum depression levels of GABA tonic
inhibition determine disease severity. Tonic inhibition is a long term form of inhibition caused by the
chronic opening of a certain type of GABA receptor that result in a continued inhibitory response.
Changes in the function of this receptor by sex steroids may implicate this channel in the depression
seen during puberty. In this project the candidate will first examine the genetic variation of the key
GABA receptor involved in tonic inhibition in both patients and controls and then compare function using
patch clamp electrophysiology. In a second series of experiments the candidate will use mouse models
we have developed that possess low, normal and high levels of tonic inhibition and analyse their
depression phenotypes using standard behavioural tests. These experiments will provide vital links
between levels of tonic current, GABA receptor function and depressive behaviour and will inform future
clinical studies.
Project 9: In vitro study of the mechanism of action of a naturally occurring
pain killer
Steven Petrou and Peregrine Osborne
Opioids are a common and effective way of treating pain but have serious side effects including
addiction, tolerance, respiratory depression and chronic constipation. Despite significant efforts in
industrial and academic laboratories, next generation, non-opioid pain medications have yet to
materialise. Recently, however, conolidine, a non-opioid natural compound isolated from the stem bark
of Tabernaemontana divaricata (Tarselli et al. 2011, Nature Chemistry 3:449) was shown to be a potent
analgesic in rodent models. In this project the candidate will undertake a series of in vitro experiments
to begin to define why this molecule has such remarkable pain killing properties. Brain slice patch
clamp electrophysiology will be used to reveal potential neuronal and synaptic mechanisms of action.
This project will be the first phase of a broader collaboration with industry in an effort to establish in vitro
and in vivo assays to test various conolidine analogues with improved drug properties and efficacy for
the treatment of persistent pain.
Project 10: Zinc and seizures
Chris Reid and Paul Adlard
Zn2+ is an essential element having a multitude of biological functions throughout the body. Febrile
seizures are common affecting approximately 3% of children. There is good evidence that febrile
seizures can trigger a cascade of events that lead to more severe forms of epilepsy later in life.
Clinically, several studies have suggested that Zn2+ levels are significantly lower in blood and CSF of
children that suffer febrile seizures but these studies are not conclusive. In this project we will directly
test the hypothesis that low brain Zn2+ may be one environmental factor in increasing the chance of
having a febrile seizure. In this project the student will learn a range of experimental techniques aimed
at understanding the role Zn2+ plays in changing neuronal excitability. The results have clear clinical
implications and could be particularly important in for developing countries, where epilepsy rates are
high and nutritional supplementation is a potential practical therapy.
Project 11: HCN channels, epilepsy and memory
Chris Reid and Marie Phillips
Humans with epilepsy often have other problems that can include memory loss, anxiety and depression.
We have data that shows that our epilepsy model learns more slowly than a non-epileptic animal. We
also know that seizures change HCN channel expression in the epilepsy mouse. HCN channels are
neuronal ion channels important for normal brain function including the ability to learn. The candidate
will investigate if changes in HCN channels are responsible for a reduction in the ability to learn in
epilepsy. The project will use whole animal behavioural studies, molecular techniques and potentially
electrophysiology to investigate this question.
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Physical Anthropology Laboratory
Dr. Varsha Pilbrow (Room E526, ph 8344 5775)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/pilbrow/
Introduction
In the Physical Anthropology Lab we use dental and skeletal morphology to address questions relating
to human biology, behaviour, health, diet and evolution. We use morphological data to study gene flow
and evolutionary diversification in fossil and ancient humans. We also reconstruct the physical
characteristics, diet, behaviour and health of past human populations. Our research materials are the
osteological collections housed in the lab, ape dental material from museums around the world, and
human skeletal material from archaeological sites.
3D scan of a human maxilla
Current projects available:
(1) PhD in ancient DNA
The project will examine the genetic diversity of ancient human populations from the region of Samtavro
and environs in the Republic of Georgia (3500 BC – 600 AD), as part of an ongoing research project on
the archaeology of the Central Caucasus. Classic and new generation DNA sequencing techniques and
SNP genotyping will be used to study familial relationships, migration and admixture. The PhD
candidate will have a strong background in molecular biology and bioinformatics, with previous studies
in biochemistry, skeletal biology, physical anthropology, archaeology, and palaeontology. The position
will be based at Melbourne University with an extended research stint at the Australian Centre for
Ancient DNA at the University of Adelaide.
(2) MSc in isotope analysis
Ratios of carbon and nitrogen isotopes will be analysed in this project to provide an understanding of
dietary behaviour and changing dietary life style in the ancient human population of Samtavro in the
Republic of Georgia (3500 BC – 600 AD). The Masters candidate will have a background in
biochemistry and anatomy. She/he will study techniques for analyzing isotopes at the Australian
National University and will liaise with isotope labs in Canberra and the University of Florida in
completing this project.
(3) MSc in discrete dental trait analysis
Dental morphological features that are non-selective in nature get fixated randomly in populations.
These discretely occurring dental traits provide an understanding of migration, admixture and genetic
relationships. In this project we will study discrete dental traits in the ancient human population from
Samtavro in the Republic of Georgia (3500 BC – 600 AD). The aim of the project is to determine the
identity of the populations represented at the burial site of Samtavro, which lies at the cross-road of
migration routes from Africa, Asia and Europe. The MSc candidate will have a background in anatomy,
with sufficient knowledge of population genetics, evolution and biostatistics. The project will necessitate
a visit to the field site in July.
43
Examples of ape and human teeth
(4) Honours in hominid taxonomy
Sample sizes for fossil hominids are typically small. To gain an understanding of patterns of variation
and evolutionary diversification in our extinct ancestors we need to compare fossil hominid variation
with variation in closely related evolutionary relatives. In this project comparative reference samples of
present-day apes will be used as models to reassess the taxonomy, or species break-down patterns, of
fossil hominids from the Miocene and Plio-Pleistocene of Africa and Europe. This lab-based project will
use scaled digital images of fossil and modern-day ape dentitions in the analysis.
Miocene ape teeth from Kenya National Museum
44
Animal Models for Cancer & Neurodegenerative Disease
Dr Leonie Quinn (Room E510A, ph 8344 5757)
Email: [email protected]
Web page: http://www.anatomy.unimelb.edu.au/researchlabs/quinn/index.html
*Project 1: Transcriptional control of the MYC oncogene
The genetic changes underlying human cancer are complex, with interactions
between multiple molecular pathways required for both disease onset and
progression. Although it has long been known that loss of cell cycle control is
extremely detrimental, with reduced cycles leading to impaired organ growth and
excessive proliferation resulting in tissue overgrowth and tumour initiation, we still
do not fully understand the precise molecular mechanisms controlling the
regulation and function of critical cell cycle genes.
The promise of Drosophila as a model continues to be the ease with which we can
use sophisticated genetics to gain novel insights into the complex molecular
mechanisms ensuring correct cell cycle patterning. Due to the high level of
conservation between the cell cycle machinery of Drosophila and humans, major
"break-throughs" have been made in our understanding of mammalian cell cycle
control. The critical role of c-MYC (simplified here as
MYC) in cancer has sparked many studies aimed to
determine the molecular and cellular basis of MYC's
oncogenic potential. The sole Drosophila member of
the family, dMyc, is functionally homologous to the MYC proto-oncogene,
and both are universally important for growth and cell cycle progression
(see figure on the left). This research aims to determine how dmyc
transcription and overgrowth is regulated in a developmental context, which
will provide critical insight into how MYC expression is dysregulated in
many human cancers; including breast, colon, lung, and prostate (for a
compilation of 93 studies see http://www.myc-cancer-gene.org).
*Project 2: Drosophila models for leukemia
Ribosomal proteins (Rps) are essential for functional ribosomes, protein synthesis, and proliferative cell
growth. Paradoxically, mutation of Rps can actually promote growth and proliferation and in some cases
bestow predisposition to cancer. Our work provided the
first rationale to explain the counter-intuitive organ
overgrowth phenotypes observed for Drosophila Rp
mutants by revealing that Rp mutants can drive tissue
overgrowth cell extrinsically, whereby reduced Rps in the
hormone-secreting gland of the larvae decreases activity
of the steroid hormone ecdysone, extending the growth
phase of development and causing tissue overgrowth (Lin
et al., PLoS Genetics 2011). This project aims to extend
these studies to better understand how Rp mutations
cause the hypoplastic anemia associated with the human
leukemia. Thus we have developed Drosophila models to
specifically reduce Rps in the hematopoietic system to
gain novel insights into how Rp mutations can promote leukemia (see lymph gland overgrowth above)
in humans. Thus we will gain further insights into the processes linking reduced levels of Rps to over
proliferation of the hematopoietic system and predisposition to cancer.
45
**Project 3: Drosophila models for neurodegenerative disease
Proteins such as Huntington can misfold and aggregate in cells to disrupt
normal cellular functioning, which is intimately associated with
neurodegenerative disease. We have developed Drosophila models for
Huntington’s disease in order to determine how defects in protein
conformation affect neural functioning. This project aims to determine the
physiological mechanisms underpinning Huntington’s disease in vivo using the
Drosophila brain (left).
Project 4: Dissecting growth regulation by novel signaling
pathways
Loss of cell cycle control is a fundamental step in cancer initiation. Crooked legs (Crol) drives cell cycle
progression and wing imaginal disc overgrowth (Mitchell et al., Development 2008), by leading to the
upregulation of critical cell cycle genes. Crol is an important upstream regulator of the Wingless (Wg)
signalling pathway. Future research is aimed towards determining how Crol regulates growth via wg
transcription.
***Project 5: The MYC oncogene in stem
cells
Cancer stem cells (CSCs) discover emphasized the
importance of interactions between stem cells and their
microenvironment or "niche". We aim to use Drosophila
stem cell models to identify the signals from the niche
required to control levels of the MYC oncogene in the
germline stem cells, which is important to prevent the
formation of germline tumours (right).
****Project 6: Defining novel roles for
ASCIZ
Since its first discovery in mice, the role of ASCIZ protein has remained elusive. Although initially
observed to participate in the DNA damage response, more recent work identified a key developmental
role during lung growth. Microarray screens revealed that expression of a number of genes are altered
in the ASCIZ-null background. Therefore, this project aims to develop Drosophila models in order to
identify novel transcriptional targets for the ASCIZ homologue (dASCIZ).
*These projects are a collaboration with Assoc. Prof. Ross Hannan at the Peter MacCallum Cancer
Centre; http://www.petermac.org/Research/GrowthControl
**This project is a collaboration with Dr Danny Hatters, Bio21
http://www.bio21.unimelb.edu.au/group-leaders/bio-chemistry/danny-hatters
Institute,
UoM;
***This project is a collaboration with 1) Dr Greg Somers, La Trobe Institute for Molecular Science
(LIMS); http://www.latrobe.edu.au/genetics/staff/GSomers/index.html and 2) Dr Gary Hime, UoM
http://www.anatomy.unimelb.edu.au/researchlabs/hime/index.html
****This project is a collaboration with Assoc. Prof. Jörg Heierhorst, St Vincents Institute, Melbourne;
http://www.svi.edu.au/research_units/molecular_genetics/unit_senior_research_staff/a_prof_joerg_heier
horst
46
Neural Regeneration Laboratory
Assoc Prof Ann Turnley ([email protected], ph 8344 3981)
Dr Kim Christie ([email protected], ph 9035 6645)
Melbourne Brain Centre
Project 1: Does traumatic brain injury (TBI) regulate hypothalamic
neurogenesis?
Traumatic brain injury can sometimes lead to altered regulation of body weight and weight is at least
partly regulated by neurons of the hypothalamus. Could altered weight regulation following brain injury
be due to altered neurogenesis in the hypothalamus? In the adult CNS, neurogenesis occurs primarily
in 2 regions, the subventricular zone (SVZ) of the lateral ventricle and the subgranular zone (SGZ) of
the dentate gyrus in the hippocampus. However, neurogenesis can also occur in the hypothalamus and
can contribute to weight regulation. We have found that there are cells in the adult hypothalamus that
label with a dye called EdU, which labels proliferative cells including neural precursor cells that produce
new neurons. This project will elucidate what cell types the EdU labelled cells are in the hypothalamus
and whether their number and type changes pre and post TBI. It will also determine whether the cell
types change depending on the severity of brain injury (mild or moderate) and whether possible
therapeutic strategies for TBI (Rho kinase inhibition and SOCS2 overexpression) alter the hypothalamic
response. This will involve use of immuno-histochemical techniques and microscopic analyses to
determine percentages of different types of EdU labelled cells – neurons (different types),
oligodendrocytes, astrocytes, microglia and inflammatory cells. Students will be provided with brain
tissue from mice with and without TBI and will also have the opportunity to participate in TBI surgery to
produce their own tissue for analysis.
Figure 1 EdU labelled cells can be detected in the adult brain (arrows point to EdU+ red
cells in right panels). Many are newborn neurons in the dentate gyrus of the
hippocampus (top right panels) but EdU+ cells are also detected in the hypothalamus
(bottom right panels). What are these cells? Dapi = nuclear stain to show all cells.
47
Cancer Biology Laboratory
Dr Elizabeth Vincan (Room E633, ph 8344 5773)
Email: [email protected] or [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/vincan/index.html
The FZD7/Wnt Signalling Pathway in Hepatocellular Carcinoma
It is estimated that at least one-third (2 billion) of the world’s population have been infected with
hepatitis B virus (HBV), of which 400 million people have chronic disease (CHB), resulting in up to 1.2
million deaths annually due to liver cirrhosis and hepatocellular carcinoma (HCC). Recently it was
shown that FZD7 plays important roles in HCC; however the effect of HBV infection in this context has
not been examined. The Cancer Biology Lab has developed powerful tools for investigating FZD7
function in colorectal cancer. We now aim to utilise these tools to investigate the interplay between
FZD7 signalling and HBV infection in HCC cells.
Specific questions to address:
1. Does modulating FZD7 signalling affect HBV infection in HCC cell lines?
We will determine the status of HBV infection in a human HCC cell line after FZD7/Wnt pathway
stimulation or inhibition.
2. Does HBV infection alter FZD7/Wnt signalling in HCC cells?
We will determine the status of FZD7/Wnt signalling in a human HCC cell line after HBV infection.
3. What are the functional consequences of the interplay between FZD7/Wnt signalling and HBV
infection in HCC cells?
To gain insight into functional effects, cell morphology and growth characteristics will be assessed.
This project will involve the analysis of fixed or inactivated material from the cell lines, for e.g.
immunohistochemistry and confocal fluorescent microscopy on human cell lines, qRT-PCR, western
blot, reporter assays. The project is suitable for an Honours project.
This project is in collaboration with Prof. Stephen Locarnini, VIDRL.
http://www.vidrl.org.au/locarnini/locarnini_professional.htm
Further reading:
Vincan E, Swain RK, Brabletz T and H Steinbeisser. Frizzled7 dictates embryonic morphogenesis:
implications for colorectal cancer progression.
Frontiers in Bioscience 12, 4558-4567 (2007)
Vincan E, Darcy PK, Farrelly CA, Faux M, Brabletz T and RG Ramsay. Frizzled-7 dictates threedimensional organization of colorectal cancer cell carcinoids.
Oncogene 26:2340-2352 (2007)
Clevers H. Wnt/beta-catenin signalling in development and disease.
Cell 127:469 (2006)
Kim M, Lee HC, Tsedensodnom O, Hartley R, Lim YS, Yu E, Merle P, Wands JR. Functional interaction
between Wnt3 and Frizzled-7 leads to activation of the Wnt/beta-catenin signalling pathway in
hepatocellular carcinoma cells.
J Hepatol. 48:780-91 (2008)
48
Autonomic Neuron Development Laboratory
Assoc Prof Heather Young (Room E524, ph 8344 0007)
Email: [email protected]
Dr Sonja McKeown (Room E505, ph 8344 5637)
Email: [email protected]
Web Page: http://www.anatomy.unimelb.edu.au/researchlabs/young/index.html
The peripheral autonomic nervous system arises from the neural crest. Our laboratory
studies the mechanisms controlling the migration of neural crest cells to and within
their target tissues, and the mechanisms controlling the differentiation of neural crestderived cells. During the development of the enteric nervous system (ENS), neural
precursors from the hindbrain must first migrate into and colonize the entire
gastrointestinal tract (gut). This migration is very interesting because (a) it takes a long
time (~3 weeks in humans and 4 days in mice), (b) the cells have to migrate very long
distances, particularly those that colonize the colon and rectum, and (c) if cells fail to
colonize the distal gut in humans, a disease called Hirschsprung’s disease results
which requires surgery.
Project 1: Live cell imaging of neural crest cell migration along the
developing gut (Primary supervisor: Heather Young)
Our laboratory has devised methods for imaging the migration of neural crest cells along explants of
embryonic mouse gut by using mice in which enteric neural crest cells express fluorescent proteins.
In this project you will use timelapse imaging and a transgenic
mouse in which neural crest cells
within the embryonic gut express
a novel fluorescent protein to
identify (a) the rules that govern
the migratory behaviour of neural
crest cells within developing gut,
and (b) the mechanisms by which
endothelin-3 and Rho GTPases
influence enteric neural crest cell migratory behaviour.
Project 2: Migration of melanoma cells in the neural crest environment
(Primary supervisor: Sonja McKeown)
This research involves placing melanoma cancer cells into the neural crest environment of chick
embryos. The neural crest are a population of cells that migrate extensively throughout the embryo and
give rise to many different cell types including bones and cartilage of the face, neurons and glia of the
peripheral nervous system and melanocytes, the pigment cells of the skin. When melanocytes become
cancerous and form a melanoma they can become invasive and migrate extensively throughout the
body. Why melanoma cells invade particular organs is currently unknown, but understanding of this
process may ultimately lead to improvements in therapy of melanoma. Previous research has shown
that melanoma cells follow the neural crest pathways when placed into chick embryos. This project
would involve placing melanoma cells obtained from different metastatic sites into the chick neural crest
pathway and determining differences in migration pattern from separate melanoma sub-populations.
The project is a collaboration with Professor Jonathon Cebon at the Ludwig Institute for Cancer
Research.
49
Project 3: Proliferation of enteric neuron precursors
(Primary supervisor: Heather Young)
Enteric neuron precursors undergo massive proliferation in order to generate the
mature enteric nervous system. In this project you will investigate the role of
various cell cycle parameters in wild-type mice and a variety of mutant mice with
defects in the enteric nervous system.
Project 4: Potential of cell therapy to treat Hirschsprung’s disease
(Primary supervisors: Heather Young and Lincon Stamp)
Infants born with Hirschsprung’s disease lack enteric neurons in the distal bowel. In this project you will
examine potential sources of cells to generate enteric neurons for use in cell therapy. Potential sources
of enteric neurons will be tested in vivo using a mouse model of Hirschsprung’s disease.
Further reading:
YOUNG, H.M., BERGNER, A.J., ANDERSON, R.B., ENOMOTO, H.,
MILBRANDT, J., NEWGREEN, D.F. and WHITINGTON P.M. (2004)
Dynamics of neural crest-derived cell migration in the embryonic
mouse gut. Dev. Biol. 270: 455-473.
HAO, M., ANDERSON, R.B., KOBAYASHI, K., WHITINGTON, P.M.
and YOUNG, H.M. (2009). The migratory behaviour of immature
enteric neurons. Dev. Neurobiol. 69:22-35.
HAO, M.M. and YOUNG, H.M. (2009) Development of enteric neuron
diversity. J. Cell. Mol. Med. 13:1193-1210.
HOTTA, R., PEPDJONOVIC, L., ANDERSON, R.B., ZHANG, D.,
BERGNER, A.J., LEUNG, J., PEBAY, A., YOUNG, H.M.,
NEWGREEN, D.F. and DOTTORI, M. (2009) Small molecule
induction of neural crest-like stem cells from human embryonic stem
cell-derived progenitor cells.
(Stem Cells, in press, accepted
16.08.09)
MCKEOWN SJ, LEE VM, BRONNER-FRASER M, NEWGREEN DF, FARLIE PG (2005) Sox10
overexpression induces neural crest-like cells from all dorsoventral levels of the neural tube but inhibits
differentiation. Dev Dyn 233:62-76.
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NOTES
51
NOTES
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