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Title: Molecular characterisation of GnRH regulated factors isolated from gonadotroph cells
Author: Chang, Lynda
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2002
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The aim of this thesis was to isolate the factors regulated by GnRH. To identify these, the LβT2 gonadotroph cell line was used as a model system for differential display RT-PCR analysis (DD-RT-PCR) of GnRH regulated transcripts. DD-RT-PCR showed that varying the GnRH pulse regime both differentially regulated and induced rapid changes in mRNA transcript levels. Myosin light chain and a putative tyrosine phosphatase were up regulated by GnRH, as were two expressed sequence tags (ESTs); one expressed in the thesis, the other in the mammary glands of pregnant or lactating mice. In addition, GnRH also down-regulated Fanconi’s anaemia complementation group A (FAA) mRNA expression levels. Fanconi’s anaemia (FA) is a autosomal recessive human disease, characterised by aplastic anaemia, short stature, developmental abnormalities, microcephaly, and infertility. Most FA genetic abnormalities map to the FAA gene, which may have a role in DNA repair, and cell cycle checkpoint control. A role for FAA in controlling gonadotrophin gene expression was tested by transient transfection assay in gonadotroph cell-lines. Transfection of a FAA expression construct specifically repressed the GnRH response of the αGSU promoter, but not the LHβ gene promoter, in LβT2 cells. The regulatory region of the FAA protein, responsible for repression of αGSU, was mapped between amino acid (aa) residues 322aa and 800aa, by serial transfection of successive C-terminal deletion constructs. Furthermore, results also suggested that FAA might mediate its effects through a paired homeodomain binding site on the αGSU promoter. Therefore, using DD-RT-PCR to isolate transcripts from GnRH treated LβT2 cells has identified novel transcripts, transcripts encoding secretion and second messenger signalling pathway proteins and FAA. Furthermore, a novel role for FAA in specifically repressing GnRH-regulated αGSU transcription was discovered.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available