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Title: Microarray expression profiling of mouse postnatal testis development and genetic models of infertility using a novel male gonadal geneset
Author: Ellis, P. J. I.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2004
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Microarray technology holds great promise for allowing the global analysis of the testicular transcriptome. The aim of this project is to demonstrate the feasibility and utility of the microarray approach to the study of the spermatogenesis. This thesis details the construction, validation and characterisation of two subtracted cDNA libraries, and their use in microarray analysis of several different models of murine-spermatogenesis. These models consist of the postnatal first wave of spermatogenesis, a series of germ-cell depleted mutant models, a series of models with deletions of the Y chromosome long arm (Yq) and the TM4 juvenile Sertoli cell line under stimulation with follicle stimulating hormone (FSH) and dihydrotestosterone (DHT). Analysis of the first wave has been used to develop a framework of expression patterns associated with different germ cell types within which to analyse other models, allowing better characterisation of the spermatogenic arrest in the germ cell depleted models. Candidate genes (including Xmr, mgclh and the unknown EST AK0059221) have been discovered with regard to the morphological abnormalities exhibited by the spermatozoa in the Yq deletion models. Microarray analysis also leads to a better understanding of the nature of the TM4 cell line, specifically how well it can be said to demonstrate the range of responses of normal Sertoli cells. Selected gene profiles of interest have been validated in this laboratory and others using established techniques of real-time RT-PCR and Northern blotting. The success of microarray technology in analysing these models demonstrates its utility for the study of spermatogenesis, and the utility of the constructed libraries in further microarray experiments. A fuller understanding of genetic events during spermatogenesis may enable therapeutic intervention in cases of male infertility, or yield insights leading to development of novel contraceptives, as well as casting light on the fundamental processes of mitosis, meiosis, fate commitment, cell differentiation and apoptosis.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available