This 30 credit module will begin with the exploration of the science of reproduction covering a range of aspects of women’s health including: normal sexual differentiation, endocrine disorders, hormonal control of fertility, pregnancy and contraception.
The module will also explore development and disease, covering embryonic development with an emphasis on molecular mechanisms and human congenital disorders. It will include an introduction to experimental techniques, terminology, model organisms and the use of transgenic mouse technology.
This pathway will take advantage of active reproduction and development research taking place at St George’s, laboratory skills sessions and clinical case-based lectures, giving you an insight into how understanding of the cellular processes involved in reproduction and development can help design strategies to aid fertility and treat/manage defects in development. The module will also provide you with insight into how new sequencing technologies and ‘omics’ methodologies are helping to decipher the cellular mechanisms involved in reproduction and development.
The substantive research project is worth 105 credits. Here are some examples of past student projects.
Does oxygen regulate stanniocalcin-1 in pregnancy?
Aims: The aims of this project are: (1) to identify which cells within the placenta express STC-1; (2) to determine whether the secretion of STC-1 by the placenta and or trophoblasts are regulated by the concentration of oxygen; and (3) to determine whether STC-1 is elevated in the serum of women in the first trimester who later develop pre-eclampsia.
Brief overview: Stanniocalcin-1 (STC1) is a glycoprotein originally discovered in bony fish where it acts to regulate calcium and phosphate homeostasis. In mammals the protein has been implicated in a number of physiological and pathophysiological processes, including ovulation, lactation, organogenesis, cerebral ischemia and tumour angiogenesis. Although in humans it is expressed by a number of tissues it has only been reported in the serum of pregnancy women and is elevated in pregnancies complicated by pre-eclampsia. Pre-eclampsia is a disorder of pregnancy characterised in late gestation by endothelial cell dysfunction, high blood pressure and proteinuria. However, the origins are believed to be in the first trimester. It is a syndrome that affects up to 8 per cent of all pregnancies, resulting in poor placental perfusion and babies that are small for gestational age. The source of the STC-1 during pregnancy is not known and therefore the regulation of its secretion has not been studied.
Methods used for data collection: Tissue and cell culture, ELISA, western blot, immunohistochemistry.
The role of calcium signalling in ciliopathy induced retinal degeneration
Aims: To use zebrafish to investigate whether aberrant calcium signalling may underlie retinal degeneration in ciliopathy patients.
Brief Overview: The cilium is a small antenna-like organelle located on the cell surface. It plays a major role in signalling pathways required for normal tissue development. There is an emerging group of human disorders, known as ciliopathies, which affect cilia function. The ciliopathies share many clinical features, including retinal degeneration leading to blindness. The photoreceptor (PR) has a modified cilium connecting the inner and outer segment, allowing transport of high-turnover light-sensitive opsins (including rhodopsin), synthesised within the cytoplasm, to the outer segment where they have their function. Cilia are enriched with ion channels required for activating the photo-transduction cascade through membrane depolarisation and stimulated neurotransmitter release. Indeed, PR death has been linked to PR overstimulation. Thus photoreceptor death might be linked to aberrant intracellular ion release. Using rhodopsin expression as readout of photoreceptor quantity, we performed an in situ hybridization (ISH) drug screen for modifiers of rhodopsin expression in zebrafish eyes using an ion channel ligand library. We found a number of calcium agonists and antagonists that affected rhodopsin gene expression. Using an eye-specific genetically encoded fluorescent calcium indicator, this project hopes to explore the function of PR cilia in relation to calcium release.
Methods used for data collection: Generation of transgenic zebrafish lines, confocal microscopy, morpholino gene knockdown, Immunohistochemistry, qPCR, zebrafish husbandry.
The effect of genome-engineered GNB1L mutations on WNT-signalling
- Generate loss-of-function mutations of GNB1L and associated proteins in HEK-293 cells
- Determine the effects of GNB1L inactivation on WNT-signalling in HEK-293 cells
Brief overview: The GNB1L gene is commonly deleted in 22q11 Deletion Syndrome (22q11DS). 22q11DS results from hemizygous deletion of a region of chromosome 22q11.2. This leads to behavioural and psychiatric problems, including autism (ASD) and schizophrenia (SZ). The study of the molecular basis of 22q11DS therefore provides a valuable opportunity to gain greater understanding of common genetic factors that cause predisposition to both ASD and SZ. GNB1L has been independently associated with ASD and SZ. Patients with schizophrenia, but without 22q11 deletion, have altered levels of GNB1L within the pre-frontal cortex. Mice that are hemizygous for GNB1L display neuro-behavioural abnormalities associated with psychiatric diseases, including schizophrenia.
GNB1L has been linked to autism, following the identification of affected individuals and families with translocation/sequence variants involving this gene. Together, these data suggest that GNB1L may be involved in the pathogenesis of psychiatric illness in 22q11DS and in the wider population. Little is known about GNB1L function, but we have discovered that it is able to regulate the WNT-signalling pathway. WNT-signalling has been previously associated with ASD and SZ. This project will further explore the cellular function of GNB1L using genome engineering techniques to create inactivating mutations of GNB1L and associated proteins in cell culture. The student will then assess the effects of these mutations on WNT-signalling.
Methods used for data collection: Cell culture, transfection, SDS-PAGE, western blotting, immunofluorescence, reporter-assays, DNA cloning, CRISPR/Cas9 genome engineering, DNA sequencing.
Molecular pathogenesis of hypogonadotropic hypogonadism
Aims: Screening of genes involved in gonadal development using zebrafish model.
Brief Overview: Congenital idiopathic hypogonadotropic hypogonadism (CHH) refers to a failure to initiate or complete puberty naturally. CHH individuals are mostly infertile due to sex hormone (LH/FSH) deficiency. Kallmann syndrome (KS) is a type of CHH, but has the additional characteristic of an absent or abnormally decreased sense of smell, known as ‘anosmia’. KS/CHH can affect both men and women, and the current treatment is the administration of GnRH or gonadotrophin to induce puberty and restore fertility. However, a significant number of patients do not respond to the current treatment and never regain normal gonadal function.
The genetic basis of KS/CHH has been well documented and mutations of different genes have been identified in around 50 per cent of cases. This project aims to investigate the pathogenesis of CHH and KS using zebrafish as a model system. We will use targeted morpholino injections and CRISPR/Cas9 technology in zebrafish to validate the impact of KS/CHH gene knockdown in the normal sexual development and function of hypothalamus-pituitary-gonadal axis, ultimately revealing new insight in the mechanisms of KS/CHH.
Methods used for data collection: In situ hybridisation, immunohistochemistry, zebrafish embryo manipulation, PCR, microscope imaging.