Inhibin: its presence, regulation and function in the zebrafish, Danio rerio.

Abstract

Poon, Shui-Kei.Thesis (M.Phil.)--Chinese University of Hong Kong, 2007.Includes bibliographical references (leaves 78-98).Abstracts in English and Chinese.Abstract (in English) --- p.iiAbstract (in Chinese) --- p.ivAcknowledgement --- p.viTable of content --- p.viiList of figures --- p.xSymbols and abbreviations --- p.xiiChapter 1 General IntroductionChapter 1.1 --- Structure of ovarian follicles --- p.1Chapter 1.2 --- Folliculogenesis and its control --- p.1Chapter 1.2.1 --- History of inhibin discovery --- p.4Chapter 1.2.2 --- Inhibin and its related proteins --- p.5Chapter 1.3 --- Inhibin --- p.6Chapter 1.3.1 --- Structure --- p.6Chapter 1.3.2 --- Function --- p.7Chapter 1.3.3 --- Signaling --- p.11Chapter 1.3.4 --- Expression --- p.14Chapter 1.3.5 --- Regulation --- p.15Chapter 1.4 --- Objectives of the present study --- p.19Chapter Chapter 2 --- Spatiotemperoal Expression Profiles of Inhibin a in the Zebrafish OvaryChapter 2.1 --- Introduction --- p.25Chapter 2.2 --- Materials and Methods --- p.27Chapter 2.2.1 --- Animals --- p.27Chapter 2.2.2 --- Chemicals --- p.27Chapter 2.2.3 --- Separation of oocytes and follicular layers --- p.28Chapter 2.2.4 --- Isolation of ovarian follicles --- p.28Chapter 2.2.5 --- RNA isolation and reverse transcription --- p.28Chapter 2.2.6 --- Real-time and semi-quantitative RT-PCR quantification of expression --- p.29Chapter 2.2.4 --- Data analysis --- p.30Chapter 2.3 --- Results --- p.30Chapter 2.3.1 --- Validation of semi-quantitative RT-PCR quantification --- p.30Chapter 2.3.2 --- Tissue distribution of inha expression --- p.30Chapter 2.3.3 --- Localization of inha expression within the ovarian follicle --- p.31Chapter 2.3.4 --- Further evidence for inha expression in the follicle layer --- p.31Chapter 2.3.5 --- Stage-dependent expression of inha in the ovarian follicles --- p.32Chapter 2.4 --- Discussion --- p.33Chapter Chapter 3 --- Regulation of Inhibin a Expression in vitro and in vivo and the Effect of Inhibin on Final Oocyte MaturationChapter 3.1 --- Introduction --- p.43Chapter 3.2 --- Materials and Methods --- p.46Chapter 3.2.1 --- Animals --- p.46Chapter 3.2.2 --- Chemicals and hormones --- p.46Chapter 3.2.3 --- Preparation of goldfish pituitary extract --- p.47Chapter 3.2.4 --- Ovarian fragment incubation --- p.47Chapter 3.2.5 --- Preparation of spontaneously matured follicles --- p.48Chapter 3.2.6 --- Intra-peritoneal injection --- p.48Chapter 3.2.7 --- RNA isolation and reverse transcription --- p.48Chapter 3.2.8 --- Real-time and semi-quantitative RT-PCR quantification of expression --- p.48Chapter 3.2.9 --- Data analysis --- p.49Chapter 3.3 --- Results --- p.49Chapter 3.3.1 --- Validation of semi-quantitative RT-PCR quantification --- p.49Chapter 3.3.2 --- Temporal change of basal inha expression in cultured ovarian fragments --- p.49Chapter 3.3.3 --- Effect of pituitary extract on the expression of inha --- p.50Chapter 3.3.4 --- Effects of recombinant zebrafish FSH and LH on inha expression --- p.51Chapter 3.3.5 --- Effect of forskolin on inha expression --- p.52Chapter 3.3.6 --- Involvement of PKA and p38MAPK in the pituitary extract and forskolin-induced up-regulation of inha expression --- p.52Chapter 3.3.7 --- Effect of recombinant zfFSH on the expression of inha in vivo --- p.53Chapter 3.3.8 --- Effects of inhibin on the basal and DHP-induced final oocyte maturation --- p.53Chapter 3.4 --- Discussion --- p.54Chapter Chapter 4 --- General Discussion --- p.71References --- p.7