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Title: Analysis of genes involved in gonadal development : identification of novel sex determination candidates
Author: Wong, F.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2005
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Genetic and developmental studies have shown that upon the presence or absence of the Y-linked sex-­determining gene SRY, the bipotential gonad will develop either as a testis or ovary. To identify new genes involved in gonadal development, a differential screen using Affymetrix GeneChip arrays was performed. Wild-type male and female gonads were dissected from mouse embryos at E12.5 (during the differentiation process) and subjected to microarray analysis. From this extensive analysis, several novel transcripts were identified, which show a sex-specific expression pattern. The validity of this approach was verified by analysing the expression of these novel transcripts by in-situ hybridisation in male and female gonads at E12.5. Transcripts with a confirmed sex-specific expression pattern where then selected for detailed in-situ hybridisation of male and female gonads at E11.5, E13.5 and E14.5. Moreover, real-time PCR of these candidates was carried out to establish a sex-specific gene expression profile during gonadal development, ranging from 17 to 36 tail somites. This thesis also describes the novel approach of applying small interfering RNAs (siRNAs) to a gonadal organ culture system. Several known sex determination genes were successfully targeted and knocked­-down by applying this technique. Importantly, siRNA against Sry effectively blocked male gonad I differentiation resulting in a lack of expression of male specific markers. Taken together these results suggest that the siRNA approach in gonad cultures can be used to efficiently analyse the function of candidate genes isolated in our sex determination screen.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available