Research Opportunities

HONORS CRITERIA

For an Honours year in the discipline of Anatomy & Histology, you need to:
- Have a Sci-WAM of at least 68*;
- pre-enrol with the Faculty of Science; organise a project with a lab head;
- confirm your intention to do Honours with the Anatomy & Histology Honours Coordinator (Prof Frank Lovicu);
- be aware of our Summer Scholarship program.

Scholarships are primarily awarded according to SciWAM. To be eligible, you must be committed to Honours in Anatomy & Histology. Application forms for Scholarships are at the end of this flyer or are available from Prof. Lovicu (Room S252).
*(If you do not meet the criteria but are still interested in Honours, see Prof. Lovicu to discuss options).

Our Honours Program in Brief
- Thesis (~20,000 words; November submission)
- Seminar (~20 minutes; November). You will present your years work.
- Honours Meetings (attend meetings, 1 hour/week during semester). At these meetings, each student will present 2 seminars during the year, outlining for example, project aims and their early results. These presentations will be to the other Honours students and the Honours Coordinators (Prof Frank Lovicu and Dr Paul Austin).

When choosing a lab to do Honours, make sure that: you get on with Supervisor; the lab is funded; the lab is filled with happy and likeable people and has recent publications; and most importantly you are interested in the project.

PROJECTS OVERVIEW

LABORATORY of NEURAL STRUCTURE & FUNCTION
Injury, Disability and Chronic Pain Research
A/Prof. Kevin KEAY (Head of Discipline)

LABORATORY OF NEUROIMMUNOLOGY AND BEHAVIOUR
PAIN AND NEUROIMMUNE CROSSTALK
Characterising the effects of neuropathic pain on appetitive motivational behaviour
Dr Paul Austin

NEUROCHEMISTRY LAB
Effects of alcohol on the expressions and regulation of glutamate transporter GLAST (EAAT1)
A/Prof. Vladimir BALCAR

ANIMAL DEVELOPMENT
Gene expression during development of the sea urchin nervous system
Prof. Maria BYRNE, Dr. Demian Koop

CEREBRAL MICROVASCULATURE AND INFLAMMATION LABORATORY
Pathology of the cerebral microvasculature - alzheimer’s disease and parkinson’s disease.
Dr. Karen CULLEN

SUNLIGHT AND CANCER GROUP
Molecular mechanisms of ultraviolet radiation-induced skin carcinogenesis
Dr Katie Dixon

CARDIAC RESEARCH LABORATORY
Chasing giant protein in failing human hearts
Prof. Cris DOS REMEDIOS

PHYSICAL ANTHROPOLOGY & COMPARATIVE ANATOMY
Human osteology with a focus on the identification of skeletal remains
Dr. Denise DONLON

ALZHEIMER'S DISEASE CELL BIOLOGY LABORATORY
Neurodegenerative disease – dissection of mechanisms in cell models and post-mortem human brain.
Dr Claire GOLDSBURY

SAVE SIGHT INSTITUTE
Study of nerve cells in the eye
A/Prof. Ulrike GRÜNERT

EYE GENETICS RESEARCH GROUP
Functional genomics and genetic eye disease
A/Prof. Robyn JAMIESON

EMBRYOLOGY UNIT
Gene function in gut endoderm development
Control of cell differentiation during mouse embryogenesis and stem cell development
Head Development: Intersection of transcriptional and signalling activities
Dr David LOEBEL, Prof. Patrick TAM

LENS RESEARCH LABORATORY
Normal Lens Biology
Lens Pathology (Cataract)

MOLECULAR NEUROBIOLOGY LABORATORY
Monoaminergic Dynamics following nerve injury
Dr David MOR

FEMALE REPRODUCTION and STRUCTURAL CELL BIOLOGY
Changes in the uterus during ovarian hyperstimulation
Prof. Chris MURPHY, Dr. Laura LINDSAY

CHILDREN'S CANCER RESEARCH UNIT, CHILDREN’S HOSPITAL: WESTMEAD
Space invaders: how cancer cells negotiate tissue barriers.
A/Prof Geraldine O’NEILL

REPRODUCTIVE TOXICOLOGY LABORATORY
Phenytoin and birth defects - Dr Helen RITCHIE
Hypoxia and changes in embryonic gene expression - Dr Helen RITCHIE, Professor Sally DUNWOODIE

REGENERATIVE MEDICINE
Skin-derived neuroprecursor cells – cell therapy and disease modeling projects
Associate Professor Michael VALENZUELA


PROJECT DETAILS

LABORATORY of NEURAL STRUCTURE & FUNCTION

Injury, Disability and Chronic Pain Research

A/Prof. Kevin KEAY | Room S502, (+61 2) 9351 4132, keay@anatomy.usyd.edu.au

LABORATORY OF NEUROIMMUNOLOGY AND BEHAVIOUR

Dr Paul Austin | Room E511, (+61 2) 9351 5061, paustin@anatomy.usyd.edu.au

PAIN AND NEUROIMMUNE CROSSTALK
Characterizing the effects of neuropathic pain on appetitive motivational behaviour

Chronic neuropathic pain in humans is associated with disabilities in complex behaviours. In particular, motivated behaviour is reduced, such that chronic pain interferes with work and social activities. In our laboratory we have characterised disabilities in social interactions and performance on the radial maze in a subgroup of neuropathic rats. Rats in the subgroup displaying behavioural disabilities have changes in dopamine and immune mediators in the nucleus accumbens, a brain region critical for the expression of motivated behaviours.


This project will characterise the effect of CCI, a common model of neuropathic pain, on appetitive motivational behavior in rats. This behavioural paradigm uses a “light-dark box”, where rats are able to receive a reward (muesli bar), but only if they leave the safe (dark) side and enter the light side, which they find aversive. By carefully analysing the specific behaviours of rats in the light-box box, and measuring the total amount of muesli consumed, we can assess their underlying motivational level. Rats will be tested in the light-dark box for 28 days (14 pre-injury and 14 post-injury), as well as sensory testing to confirm the presence of neuropathic pain. From our previous studies we know CCI causes a range of behavioural disabilities across a population of rats, despite equal levels of pain. Therefore we predict some rats will show disabilities in appetitive motivational behaviour after injury. The project will also involve immunohistochemical analysis of brain tissue collected from behaviourally tested rats to examine correlations with neuroimmunological changes in the brain.

NEUROCHEMISTRY LAB

A/Prof Vladimir BALCAR | Room S419, ext 12837, vibar@anatomy.usyd.edu.au          

Effects of alcohol on the expressions and regulation of glutamate transporter GLAST (EAAT1)

The second most abundant glutamate transporter in the CNS is EAAT1 (GLAST). It is located in astrocytes where it acts synergistically with the most commonly expressed glutamate transporter GLT1 transporting glutamate away from the extracellular space into the astrocytes. Unlike GLT1, GLAST transporter seems to form transient and rather labile complexes with Na+/K+-ATPase. In addition, it shifts rapidly between the cytoplasm and plasma membrane depending on the demand so that it can rapidly respond to changes in glutamate concentrations outside the cell (Bauer et al 2012, Šerý et al 2015).

We have found that alcohol disrupts the orderly shifts of GLAST from cytoplasm to the plasma membrane (Balcar et al 2015; Šerý et al 2015) thus precluding the effective formation of GLAST and Na+/K+-ATPase complexes and impairing the GLAST function. In vivo, this would disturb the fine regulation of glutamate transport eventually leading to permanently increased extracellular glutamate (“hyperglutamatergic” state, cf. Spanagel et al. 2005). Therefore, the voluntary self-administration of alcohol (drinking) would provide less relief of craving unless the dose keeps going up (for a review see Spanagel 2009). There may be a long-term but, because of the disruption of the above regulatory process, less and less effective compensation by overexpression of GLAST (Flatcher-Bader and Wilce 2008). The beginnings of such process might be observable in our model if we expose the cultured astrocytes to alcohol for long enough time (hours or days) and measure the total expression of GLAST by Western blotting. We can also follow the movements of GLAST in and out of the plasma membrane in the presence of glutamate (or a more effective GLAST substrate D-aspartate) with and without alcohol. The latter methodology is based on a combination of immunocytochemistry using and deconvolution microscopy as available in our Department (Shin et al 2009, Nguyen et al 2009; 2010). GLAST antibodies have been supplied by Prof. David Pow (RMIT, Melbourne).

We have also been looking at the expression of GLAST (as well as other relevant proteins) in human alcoholic brains.

  • Balcar VJ, Taylor DL, Sultana N, Kashem MA, Pow DV (2015) Ethanol causes translocation of GLAST from cytoplasm to plasma membrane: the effect is blocked by baclofen. J Neurochem S1, (Abstr. No. WTH01-02) 243
  • Bauer DE, Jackson JG, Genda EN, Montoya MM, Yudkoff M and Robinson MB (2012) The glutamate transporter GLAST participates in a macromolecular complex that supports glutamate metabolism. Neurochem International 61 566-574; doi: 10.1016/j.neuint.2012.01.013
  • Flatcher-Bader T and Wilce PA (2008) Midkine and EAAT1 in the human prefrontal cortex Alcoholism: Clin Exp Res 32 1849-1858
  • Nguyen KTD, Buljan V, Pow DV and Balcar VJ (2010) Cardiac glycosides ouabain and digoxin interfere with the regulation of glutamate transporter GLAST in astrocytes Neurochem Res 35 2062-2069
  • Nguyen KTD, Shin JW, Rae C, Nanitsos EL, Acosta GB, Pow DV, Buljan V, Bennett MR, Else PL and Balcar VJ (2009) Rottlerin inhibits (Na+,K+)-ATPase activity in brain tissue and alters D-aspartate dependent redistribution of glutamate transporter GLAST in cultured astrocytes Neurochem Res 34 1767-1774
  • Å erý O, Sultana N, Kashem MA, Pow, DV and Balcar VJ (2015) GLAST but not least – distribution, function, genetics and epihenetics pf L-glutamate transport in brain – Focus on GLAST/EAAT1 Neurochem Res doi: 10.1007/s11064-015-1605-2
  • Shin JW, Nguyen KTD, Pow DV, Knight T, Buljan V, Bennett MR and Balcar VJ (2009) Distribution of glutamate transporter GLAST in membranes of cultured astrocytes in the presence of glutamate transport substrates and ATP Neurochem Res 34 1758-1766
  • Spanagel R (2009) Alcoholism: A system approach from molecular physiology to addictive behaviour Physiol Rev 89 649-705
  • Spanagel R et al., (2005) The clock gene Per2 influences the glutamatergic system and modulates alcohol consumption. Nature Medicine 11 35-42
ANIMAL DEVELOPMENT

Prof. Maria BYRNE | Room S600, (+61 2) 9351 5166, mbyrne@anatomy.usyd.edu.au

Dr. Demian Koop | demian.koop@sydney.edu.au

Gene expression during development of the sea urchin nervous system

Sea urchins are a leading model for understanding the genetic networks that regulate animal development. Our lab uses Australian sea urchin species to investigate how changes to development have led to the evolution of the unique “five-rayed” organization or the sea urchin body. This will allow us to establish which parts of the genetic blueprint for development have been conserved between sea urchins and vertebrates, and will provide insights into the vertebrate ancestor.

Our current ARC funding has allowed us to establish the developmental transcriptome of two local sea urchins, which we are using to investigate the genes involved in many aspects of development including axial patterning – such as Anterior-Posterior and Left-Right, genes involved in skeleton formation and genes involved in the development of the central nervous system. The projects will involve using the transcriptome to identify and characterise the genes involved in these aspects of development. This will involve expression analysis using in situ hybridization and confocal microscopy to map gene expression domains during development. The role of key signaling pathways in skeletogenesis and neurogenesis will be determined by analyzing the effects of pharmacological treatments on development of sea urchin embryos.

CEREBRAL MICROVASCULATURE AND INFLAMMATION LABORATORY

Dr. Karen CULLEN | Room S464, (+61 2) 9351 2696, kcullen@anatomy.usyd.edu.au

Pathology of the cerebral microvasculature - alzheimer’s disease and parkinson’s disease
The microvasculature: degeneration, inflammation and Alzheimer’s disease

Our laboratory is focused on pathology of the cerebral microvasculature. The brain has an extremely dense and very complex microvasculature. Health of this vascular network is essential for normal brain function. Several neurological conditions are marked by pathology of these tiny vessels, including Alzheimer’s disease (AD), Parkinson’s disease (PD) and head injury. Our work investigates the anatomy of the normal and diseased microvasculature, as well as the relationship of damaged vessels to neurodegeneration.

Several projects are offered in our laboratory. For instance, the key lesion in the AD is the breakdown of the cerebral microvasculature. Projects in AD involve the mapping of capillary damage and the sequence of inflammatory events from fresh microhaemorrhage to scar formation. In PD, we find breakdown of the microvessels in the terminal fields of the substantia nigra neurons, the cells that die in this movement disorder. Our PD project aims to map vascular anomalies in the striatum in human brain tissue. The techniques used in the lab include tissue processing, immunohistochemistry, brightfield and fluorescence microscopy. We use multidimensional analysis to examine the complex vascular network in human brain tissue.

These projects would suit high achieving motivated students with some background in neuroanatomy and good literature research and writing skills.

SUNLIGHT AND CANCER GROUP

Dr. Katie DIXON | Rm. S228, (+61 2) 9351 4633, katie.dixon@sydney.edu.au

Skin cancer is highly prevalent in Australia, with two in three people being diagnosed by the age of 70. While non-melanoma skin cancers are more common, melanoma is responsible for the majority of deaths related to skin cancer. Projects in this laboratory involve the investigation of the molecular mechanisms of ultraviolet radiation-induced skin carcinogenesis, as well as inhibition of the growth and metastasis of melanoma.

Interests outside of skin cancer include investigation of cell signaling pathways in cancer, with an emphasis on identification and targeting of tumour suppressor genes.

Techniques include but are not limited to cell culture, immunohistochemistry, western blotting, RT-PCR, in vivo studies and simulation of ultraviolet radiation.

CARDIAC RESEARH LABORATORY (formerly the Muscle Research Unit)

Prof. Cris DOS REMEDIOS | Room S468. (+61 2) 93513209, cris.dosremedios@sydney.edu.au

Chasing giant protein in failing human hearts

The most common cause of human heart failure is Idiopathic Dilated Cardiomyopathy (IDCM), and the most common protein defect is a mutation in the giant protein titin. Titin is the largest known protein with a molecular weight of nearly 4 million Daltons. We have evidence (just accepted for publication in PLoS One) that a different giant protein is also involved in IDCM.

This honours project will establish the molecular basis for the action of this second, previously unidentified mutated protein using Western blots and qRT PCR using heart samples from IDCM hearts in the Sydney Heart Bank.

PHYSICAL ANTHROPOLOGY & COMPARATIVE ANATOMY

Dr. Denise DONLON | Room W601, (+61 2) 9351 4529, ddonlon@anatomy.usyd.edu.au

Research in the Shellshear Museum focuses on human osteology with a focus on the identification of skeletal remains.  Present research focuses on discriminating between human and non-human bones, taphonomic changes to bone, the anthropometry of early British settlers in Australia and age determination using amino acid racemization of tooth enamel. Collections in the Shellshear Museum, which are available for research, include a large collection of Melanesian skulls, a collection of skeletal remains from Pella, Jordan and casts of human dentition.

ALZHEIMER'S DISEASE CELL BIOLOGY LABORATORY

Dr. Claire GOLDSBURY BMRI, (+61 2) 9351 0878, claire.goldsbury@sydney.edu.au

Neurodegenerative disease – dissection of mechanisms in cell models and post-mortem human brain.

Using cell models and post-mortem human brain tissue, this lab investigates the interactions between tau, cofilin and other proteins that are involved in generating pathological structures involved in neurodegeneration and dementia. This work will help us to understand how these structures could contribute to neurodegeneration and the resulting cognitive decline that characterizes Alzheimer's and other neurodegenerative disease.

SAVE SIGHT INSTITUTE

A/Prof. Ulrike GRÜNERT | Save Sight Institute, (+61 2) 9382 7641, ulrike.grunert@sydney.edu.au

Study of nerve cells in the eye

The retina is a thin piece of nervous tissue that lines the back of the eye. It contains five major classes of neurones, which are organised in nuclear and synaptic layers. The project involves the analysis of post-mortem human retinas (obtained from donor eyes with consent from the Lions NSW Eye Bank) and post-mortem monkey retinas. High-resolution light microscopy, in conjunction with immunohistochemical methods will be used for identifying different cell types and their synaptic connectivity. Accurate knowledge of nerve cell populations and their density can allow more accurate prediction of functional properties of normal visual performance, can give a rational basis for understanding the effects of disease and degeneration, and can form a standard of normal anatomical state against which the effects of dis- ease can be measured.


Skills and knowledge you will acquire
We are looking for an enthusiastic young scientist to learn neuroanatomical methods including cryostat and Vibratome sectioning of retinas, intracellular injections, double and triple immunofluorescence, high resolution light microscopy (deconvolution and confocal) and image analysis.

EYE GENETICS RESEARCH GROUP

A/Prof. Robyn JAMIESON | Children’s Medical Research Institute - The Children’s Hospital at Westmead, and Save Sight Institute, (+61 2) 9687 2800, rjamieson@cmri.org.au

Functional genomics and genetic eye disease

We use genomic, cell-based and animal model studies to investigate genes and pathways that are critical for normal sight, and that are abnormal in patients with blinding eye conditions such as retinitis pigmentosa, glaucoma, or congenital cataracts. These investigations are fundamental in our work towards treatment of these blinding eye disorders.

Projects available in the lab may include one or a combination of the following areas. Whole exome, whole genome and other forms of next-generation sequencing (NGS), along with the bioinformatics approaches for analysis of the data, are used to identify candidate causative disease genes. Specific functions of disease genes are investigated using cell-based and mouse model studies.

Available techniques include bioinformatic analysis and interpretation of NGS data, morphological and histological examination, immunohistochemistry, gene expression microarray and RNA-Seq, qRT-PCR, gene editing using CRISPR/Cas9, and cell-based assays.

  • Greenlees R, Mihelec M, Yousoof S, Speidel D, Wu SK, Rinkwitz S, Prokudin I, Perveen R, Cheng A, Ma A, Nash B, Gillespie R, Loebel DA, Clayton-Smith J, Lloyd IC, Grigg JR, Tam PP, Yap AS, Becker TS, Black GC, Semina E, Jamieson RV. Mutations in SIPA1L3 cause eye defects through disruption of cell polarity and cytoskeleton organization. Human Molecular Genetics, 2015 Jul 30. [Epub ahead of print] PMID: 26231217
  • Nash BM, Wright DC, Grigg JR, Bennetts B, Jamieson RV.  Retinal dystrophies, genomic applications in diagnosis and prospects for therapy. Translational Pediatrics. 2015; 4(2): 139-163.
  • Prokudin I, Li D, He S, Guo Y, Goodwin L, Wilson M, Rose L, Tian L, Shen Y, Liang J, Keating B, Xu X, Jamieson RV, Hakonarson H. Value of whole exome sequencing for syndromic retinal dystrophy diagnosis in young patients. Clinical and Experimental Ophthalmology. 2015 Mar;43(2):132-8. PMID: 25060287
EMBRYOLOGY UNIT

Dr David LOEBEL  | Children’s Medical Research Institute - Westmead, (+61 2) 8865 2953, dloebel@cmri.org.au

Prof. Patrick TAM | Children’s Medical Research Institute - Westmead, (+61 2) 8865 2800, ptam@cmri.org.au

Gene function in gut endoderm development.

This project addresses a fundamental issue of embryonic development at the start of life: the molecular activity controlling the formation of major body parts of the embryo. The foregut forms the liver, pancreas, the epithelium of the digestive tract and lungs, the thymus, thyroid and parathyroid glands. The molecular basis for the formation, organization and differentiation of these organs is not well understood, and this project is aimed at contributing to our knowledge of this process. We are using a variety of approaches to study the functions of some of these genes during development of the endoderm and its derivative organs. In this project, the effects of reduced or loss of gene function (by knockdown, gene-targeting or gene-trap) and gain of gene function (by electroporation, transfection and transgenesis) will be tested in mouse embryos, embryonic stem cells and other appropriate cell models.

Control of cell differentiation during mouse embryogenesis and stem cell development.

The knowledge of how to maintain, expand and differentiate stem cells is essential for the realization of clinical cell-based therapy for the replacement and repair of diseased tissues.  Cells of the early embryo are capable of generating many cell types but as the embryo develops, there is a progressive restriction of the ability of the cells to do so.  We are documenting the gene expression profiles of the cell populations at throughout the embryo, to track the activity of the gene regulatory networks driving cell fate choices.  The network and pathway function will be verified by tracking lineage differentiation in genetically modified embryos.

Head Development: Intersection of transcriptional and signalling activities.

In humans, lethal malformation complexes of the head are associated with varying degrees of anatomical defects of the brain, skull and face structures. It is believed that these major anatomical defects of the craniofacial structures result primarily from abnormal development in the first trimester of human development. This coincides with the time window of head formation, 7-11 days after conception in the mouse embryo, which is utilized as an experimental model for analysing the genetic and developmental mechanisms of the pathogenesis of the human malformations.  This project focuses on analysing the genetic and molecular activities that control the formation of the embryonic head and will utilize genetic and embryological models in which transcription factor coding genes are ablated in a tissue-specific manner to study the impact on the severity of head malformation. We will also analyze the connection of these transcription factors with WNT signalling activity thatinfluences tissue differentiation and cell movement in the formation of the embryonic head and face.

LENS RESEARCH LABORATORY

Prof. Frank LOVICU | Room S252, (+61 2) 9351 5170, frank.lovicu@sydney.edu.au

Research in our laboratory is directed at identifying the molecular mechanisms that regulate eye lens development, growth and pathology. Using a range of techniques (including tissue culture, immunohistochemistry, in situ hybridisation, PCR, chromatography, Western blotting, light and electron microscopy, in vitro biological assays and transgenic mouse strategies), we investigate the expression, effects and function of different growth factors and their receptors as well as the regulation of their intracellular signalling, both in normal lens development and pathology. To date, we have shown that members of the fibroblast growth factor (FGF) and Wnt families are important regulators of lens epithelial cell proliferation, migration and differentiation and are important for the normal development and maintenance of the lens. Our other studies have also shown that growth factors such as transforming growth factor ß (TGF-ß), induce the formation of fibrotic plaques that lead to cataract (loss of lens transparency), similar to that found in humans.

Students that undertake Honours projects in our laboratory can expect to be exposed to a wide array of techniques, encompassing cellular, developmental and molecular biology, and can carry out a project in one or a combination of the following areas:

Normal Lens Biology

  • Investigate the role of growth factors (FGF, PDGF, IGF, EGF, BMPs) and their signalling pathways in regulating lens cell proliferation and fibre differentiation using lens epithelial explants and/or transgenic mouse models.
  • Identify factors that maintain the normal lens epithelial phenotypic characteristics including cell-cell and cell-matrix adhesion and communication.
  • Use transgenic mice and in vitro assays to determine the role of novel genes (Crim1, Sef, Sprouty1/2, Spreds1/2/3) thought to be involved in regulation of growth factor bioavailability and signalling.
  • Use electron microscopy and tissue culture to identify the molecules in the ocular fluid that are important for lens cell differentiation and how this contributes to lens transparency.

Lens Pathology (Cataract)

  • Use transgenic mouse models to understand how TGFß induces and regulates cataract formation.
  • Use lens explant cultures to determine how TGFß disrupts normal lens signalling pathways and induces an epithelial-mesenchymal transition, characteristic of cataract.
  • Use lens explant cultures to identify putative inhibitors of TGFß signalling as a means of preventing cataract.
MOLECULAR NEUROBIOLOGY LABORATORY

Dr David MOR | david.mor@sydney.edu.au

Monoaminergic Dynamics following nerve injury

A significant proportion of patients who have suffered traumatic nerve injury, following accident or surgery, subsequently experience disability, characterised by anxiety, depression and disturbances in cognition and sleep cycles.

The main focus of this lab is on the alterations caused by the nerve injury within the neural network that regulates behavioural and physiological responses to stress and pain. We specifically study differences in serotonergic and dopaminergic neurotransmissions from selected midbrain and brainstem nuclei into two brain regions within this network: the nucleus accumbens and the prefrontal cortex (PFC). The nucleus accumbens is important for regulation of motivational drive. The PFC is considered an executive regulator of the coping responses. Its function is to incorporate and assess information from the environment and responsible for decision making of which behaviour to produce.

The project involves identifying individual differences in the regulation these monoaminergic projections that are specific to the expression of disability following a nerve injury. Techniques used will include behavioural testing of rats following a nerve injury, molecular techniques analysing epigenetic differences as well as gene and protein expression, and immunohistochemistry.

FEMALE REPRODUCTION and STRUCTURAL CELL BIOLOGY

Prof. Chris MURPHY | Room N364, (+61 2) 9351 4128, histology@medsci.usyd.edu.au

Dr. Laura LINDSAY | Room N364, (+61 2) 9351 2508, laural@anatomy.usyd.edu.au

The work in this lab is centred around reproductive biology and medicine and in particular the biology of the uterus, uterine receptivity for blastocyst implantation and hormonal influences on the uterus. We are interested in how it is that the uterus manages to tightly regulate those times during the reproductive cycle when it will allow the blastocyst to attach but to prevent attachment and the beginning of a pregnancy at other times. We are particularly interested in uterine epithelial cells and the molecular interactions that occur between the surface of these cells and the implanting blastocyst.

A variety of methods are available including light & electron microscopy, immunohistochemistry, Western blotting and PCR. The work uses both animal and human tissues and involves basic cell biological research as well as work on human tissues of direct relevance to the human menopause and to In vitro fertilisation (IVF) programmes. The laboratory also has extensive contacts with The School of Biological Sciences and the Electron Microscope Unit (EMU) which involves a major project on the evolution of viviparity (live birth) and the development of the placenta. This work involves study on mammals and lizards in particular but also other animals to understand the biology of different types of placentas.

We also have an interest in one of the major diseases of the uterus which affects over a million Australian women endometriosis - and have collaborations with Westmead hospital to study this disease. We would accept students interested in mammalian reproduction and/or students interested in working on an aspect of the evolution of live birth and placentation. An honours place in conjunction with the EM Unit could also be arranged. We are particularly interested in accepting Honours students who may be interested in progressing to a PhD and have laboratory-funded top-up scholarships available for such students.

 Changes in the uterus during ovarian hyperstimulation

Controlled ovarian hyperstimulation (COH) is part of the in vitro fertilisation (IVF) protocol to harvest multiple eggs from a woman. It has been known for many years that the drugs used in COH leads to a decrease in uterine receptivity, however there is currently no known mechanism of how this occurs. We use a rat ovarian hyperstimulation model (OH) to investigate changes in the rat uterus during OH pregnancy, especially at the time of blastocyst implantation. We use several molecular and histochemical techniques to investigate changes in protein localisation and expression, such as immunofluorescence microscopy, western blotting and ELISAs. We also use a range of microscopy techniques such as light microscopy, transmission electron microscopy and scanning electron microscopy.

The ultimate aim of this project is to identify factors or proteins associated with the decreased uterine receptivity in women undergoing ovarian hyperstimulation. A more detailed understanding of these factors may increase the pregnancy rate in IVF procedures.

CHILDREN'S CANCER RESEARCH UNIT, CHILDREN’S HOSPITAL: WESTMEAD

A/Prof Geraldine O’NEILL | Children's Cancer Research Unit - Children’s Hospital at Westmead, geraldine.oneill@health.nsw.gov.au

Space invaders: how cancer cells negotiate tissue barriers.

For many years, tumours have been diagnosed by the presence of a lump that can be detected by touch. This ability to palpate the tumour is due to increased rigidity of the tumour matrix. More recently, research in the tissue engineering field has revealed that matrix rigidity is an important determinant of cell fate. These two observations have now come together, in a growing recognition that external mechanical forces regulate cancer cell invasion. This realization means that investigation of cancer cell migration and invasion in flat, plastic dishes does not provide a faithful replicate of the in vivo tissue environment. The dishes are significantly more rigid than any tissues in the body and they do not mimic the 3-dimensional (3D) in vivo tissue structure that provides a barrier to cell invasion.

In order to investigate how mechanical forces regulate cancer cell invasion, our lab therefore employs a range of cell culture models that mimic in vivo tissue and tumour organization, with a focus on brain cancer and neuroblastoma.

We have projects available to investigate:

  • how the 3-dimensional (3D) biophysical environment regulates cancer invasion;
  • how external tissue rigidity effects cancer cell response to chemotherapy; and
  • how actin networks control cell movement in 3D environments. Techniques employed include fluorescence microscopy, time-lapse microscopy, molecular biology and biochemistry.
REPRODUCTIVE TOXICOLOGY LABORATORY

Dr Helen RITCHIE | Room S449, (+61 2) 9351 9476, helen.ritchie@sydney.edu.au

Phenytoin and birth defects

Our laboratory is interested in why some drugs taken during pregnancy cause birth defects. We use rat models to answer this question. Phenytoin is a drug used to control epilepsy. However it causes cleft lip in humans if taken during pregnancy. The mechanism by which phenytoin causes birth defects remains unknown but there is a strong hypothesis that phenytoin that crosses the placenta slows the embryonic heart causing the fetus to become hypoxic for some hours. If this hypoxia occurs during critical periods of facial development it is thought to induce abnormal development.

We are going to test this hypothesis by examining the heart rate of the embryo in vivo by ultrasound and in vitro by embryo culture. There is an alternate hypothesis which proposes that the malformations are caused by hyperglycaemia. We will test this hypothesis by measuring the glucose levels in pregnant rats following a teratogenic dose of phenytoin.

 

Dr Helen RITCHIE | Room S449, (+61 2) 9351 9476, helen.ritchie@sydney.edu.au

Professor Sally DUNWOODIE | Victor Chang Cardiac Research Institute, s.dunwoodie@victorchang.edu.au

Hypoxia and changes in embryonic gene expression

Our laboratory is interested in the causes of birth defects. We use animal models to address this issue. Dofetilide is a drug used to control human cardiac arrhythmia. In laboratory animals dofetilide causes birth defects, thus there is a risk that it might cause similar malformations in humans if taken during pregnancy. In rats we have shown that dofetilide slows the fetal heart and causes arrhythmia, leading to fetal hypoxia for some hours (Ritchie et al 2013). We have also established that mouse embryos exposed to 8 hours of hypoxia, half way through gestation, develop defects of the vertebral column (Sparrow et al 2012).

We hypothesis that hypoxia induced by dofetilide, will lead to vertebral column defects in mice. To test this hypothesis we will administer dofetilide to pregnant mice and determine the extent to which dofetilide causes hypoxia in embryos. Embryo hypoxia will be determined using two markers, hypoxyprobe (via western blot) and HIF1a expression (via in situ immunofluorescence). We will also examine the effect of dofetilide on the heart rate of embryos exposed to dofetilide in vitro by embryo culture. We have shown that gene-environment interaction causes developmental defects in mice (Sparrow et al 2012). Specifically, embryos genetically susceptible to vertebral column defects develop such defects, when exposed to hypoxia during gestation. We will test if similar interaction occurs between these genetically susceptible embryos and the hypoxia induced by dofetilide.

REGENERATIVE MEDICINE

Associate Professor Michael Valenzuela | BMRI - Building K Level 4, (+61 2) 9114 4136, michael.valenzuela@sydney.edu.au

Skin-derived neuroprecursor cells – cell therapy and disease modeling projects

Neural stem cells (NSCs) can divide indefinitely and under controlled conditions differentiate into the brain’s three main cell types – neurons, glia and oligodendrocytes. Currently, there are three main strategies for obtaining NSCs: from the brain’s neurogenic niches that persist into adulthood, from culture of embryonic stem cells or from genetic engineering of embryonic stem cell-like induced pluripotential cells.

Each of these approaches present non-trivial challenges for clinical application and in vitro disease modeling. Our laboratory has therefore developed a protocol for generating skin-derived neural precursors (SKNs) from a small biopsy of mature adult skin. SKNs are most likely of neural crest origin and are both rate- and fate- restricted, capable of 3-5 cells divisions and differentiate almost exclusively into neuron. In culture, SKNs have a strong gabaergic bias that stimulates interest in modeling of neuropsychiatric disorders. SKNs also survive, migrate, elaborate dendrites and display network functionality after hippocampal transplantation in an ageing rodent model, resulting in rescue of memory dysfunction. Honors projects will continue to progress this research and combine both in vitro and in vivo neuroscience techniques.