For information about current projects or to suggest your concept for a project, please contact one of our research group leaders. All are happy to meet with potential honours students. The following is a sample of the honours projects on offer at ARMI.
Characterisation of a novel dystrophic zebrafish mutant
Zebrafish models of disease are particularly valuable because zebrafish embryos develop ex utero, are optically clear, highly manipulable, and genetically tractable (Lieschke and Currie, Nat Rev Genet 2007).
In a zebrafish ENU-screen performed in our laboratory, we isolated several novel mutants with dystrophic muscle phenotypes. These mutants show muscle fibre degeneration and loss of birefringency as depicted below.
The aim of the proposed project is to characterise the muscle defects of one of the novel zebrafish mutants and a to perform a first-pass-mapping.
In our newly built, state-of-the-art fish-facility, basic experiments like survival curves and breeding schemas for initial mapping crosses will be carried out. Immunohistochemistry together with confocal microscopy and histological staining techniques with will be utilised to survey the molecular character of the mutant muscle at different embryo and larvae stages.
Supervisor: Dr Joachim Berger
Imaging leukocyte function in infection and inflammation
Zebrafish models of infection and inflammation offer unrivalled opportunities to observed white blood cell function during infection and inflammation in a whole living animal. We have transgenic zebrafish expressing fluorophores in different white blood cell types, and can study the migratory and functional behaviour of these cells during sterile inflammation or experimental infection. Observing these processes for the first time in vivo is revealing previously unexpected phenomena. This project aims to quantitatively image neutrophil and macrophage behaviour in a zebrafish model of sterile inflammation with regeneration, and to determine how this behaviour is affected in a zebrafish mutant with myeloperoxidase deficiency. Myeloperoxidase deficiency is one of the commonest disorders of human leukocyte function. The project will combine genetic tools with sophisticated high-end imaging and computerised image analysis to better understand a basic pathological process.
Supervisor: Professor Graham Lieschke
Med12, a transcriptional regulator important in early blood cell development
Haematopoiesis requires the spatiotemporal organization of regulatory factors to successfully orchestrate diverse blood cell lineage specificity from stem and progenitor cells. Med12 is a regulatory component of the large Mediator complex that enables contact between the general RNA polymerase II transcriptional machinery and enhancer bound regulatory factors. We have identified a new zebrafish med12 allele, syrah, which has defects in hematopoiesis, predominantly in the myeloid lineage, suggesting a hematopoietic cellspecific requirement for Med12.
Identification of blood-specific interacting partners for Med12: Med12 has a protein-protein interacting domain and has been shown to bind proteins important in neural development. This project will use a two-pronged approach to identify leukocyte-specific Med12 interacting partners:
- the classic yeast two-hybrid screen; and
- a range of cell and molecular biology techniques including coimmunoprecipitation, immunoblotting, and cutting-edge LC-MS/MS mass spectrometry. The identification of blood-specific interacting proteins will direct future studies targeting the myeloid cell-specific function of Med12.
Supervisor: Dr Cristina Keightley
Biochemistry of Zbtb11, a novel transcriptional regulator
Marsanne is a mutant with a defect in myeloid development, which maps to Zbtb11. Zbtb11 is a transcriptional repressor required for normal embryonic development and white blood cell development in zebrafish. There is evidence suggesting that like other BTB/POZ proteins, the importance of Zbtb11 in normal blood cell production underpins its dysregulation and potential role in human blood cancers.
- Downstream targets of Zbtb11: This project will use a range of cutting-edge molecular biology techniques including microarrays to determine which genes are controlled by Zbtb11. Potential target genes will then be explored using in situ hybridization and RT-PCR. Together, these studies will provide a handle on the gene networks by which Zbtb11 regulates myelopoiesis.
- Phosphorylation of Zbtb11: Phosphorylation is an important mechanism by which transcription factors can be switched on/off or modulated in response to specific stimuli. There is evidence suggesting that Zbtb11 is a phospho-protein. This project will use a range of cell and molecular biology techniques including site-directed mutagenesis, cell culture, reporter assays and a zebrafish in vivo bioassay to ascertain which phosphorylation sites are important for the function of Zbtb11.
- Requirements for Zbtb11 in development and identification of upstream genetic
partners of zbtb11: This project will take advantage of the temperature sensitivity of the marsanne allele and use it as a conditional tool to determine the onset of Zbtb11 requirement in development. In addition, a transgenic Zbtb11-reporter zebrafish line will be created in order to mark Zbtb11-expressing cells, and other myeloid-defective mutants in the laboratory will be screened for aberrations in Zbtb11 expression by whole mount in situ hybridization (WISH), indicating an upstream regulatory role in Zbtb11 function. Cell culture and reporter assays will also be used to determine regulation of the Zbtb11 promoter by candidate upstream factors.
The data generated in this project will provide important insight into the role of Zbtb11 in mammalian blood cell regulation.
Supervisor: Dr Cristina Keightley
Investigating the role of microRNAs in vertebrate development
Patterning of the early vertebrate embryo requires precise spatial and temporal control of Hox gene expression during embryonic development. While this is largely achieved through extensive regulatory mechanisms at the level of gene expression, we have identified an additional layer of regulatory control provided by the miR-196 family of microRNAs in defining the boundary of Hox gene expression along the anterior-posterior embryonic axis. We have generated knock-out mice for individual miR-196 family members to understand the impact of these tiny RNA species during embryonic development.
This project will aim to investigate the importance of miRNA control during the formation of particular organ systems, and will capitalise on the unique knockout resources already established within the lab. This project will utilise numerous developmental and molecular techniques such as in situ hybridisation, immunofluorescence, FACS analysis and quantitative genome analysis approaches.