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Hormonal regulation of vitellogenin expression in the goldfish.January 2002 (has links)
Pang Yee Man Flora. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 111-128). / Abstracts in English and Chinese. / Abstract (in English) --- p.ii / Abstract (in Chinese) --- p.iv / Acknowledgement --- p.v / Table of Contents --- p.vii / List of Figures --- p.xii / Symbols and Abbreviations --- p.xv / Scientific Names --- p.xvii / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- Vitellogenesis --- p.2 / Chapter 1.2 --- Vitellogenin --- p.3 / Chapter 1.2.1 --- Structure --- p.3 / Chapter 1.2.2 --- Vitellogenin synthesis in the liver --- p.4 / Chapter 1.3 --- Regulation of vitellogenin synthesis --- p.5 / Chapter 1.3.1 --- Estradiol --- p.5 / Chapter 1.3.1.1 --- Mechanism of action --- p.6 / Chapter 1.3.1.2 --- Estradiol-stimulated vitellogenin expression --- p.7 / Chapter 1.3.1.3 --- Memory effects --- p.9 / Chapter 1.3.2 --- Testosterone --- p.10 / Chapter 1.3.3 --- Cortisol --- p.13 / Chapter 1.3.4 --- Progesterone --- p.14 / Chapter 1.3.5 --- Growth Hormone --- p.14 / Chapter 1.3.6 --- Prolactin --- p.15 / Chapter 1.3.7 --- Thyroid hormone --- p.15 / Chapter 1.4 --- Growth factors --- p.16 / Chapter 1.4.1 --- Activin --- p.16 / Chapter 1.4.1.1 --- Structure --- p.16 / Chapter 1.4.1.2 --- Functions --- p.17 / Chapter 1.4.2 --- Epidermal growth factors (EGF) --- p.18 / Chapter 1.4.2.1 --- Structure --- p.18 / Chapter 1.4.2.2 --- Functions --- p.19 / Chapter 1.5 --- Objectives of the present study --- p.20 / Chapter Chapter 2 --- Expression of Goldfish Vitellogenin in vivo and in vitro --- p.25 / Chapter 2.1 --- Introduction --- p.25 / Chapter 2.2 --- Materials and Methods --- p.26 / Chapter 2.2.1 --- Materials --- p.26 / Chapter 2.2.2 --- Sequencing --- p.27 / Chapter 2.2.3 --- Cell culture --- p.28 / Chapter 2.2.4 --- RNA extraction --- p.29 / Chapter 2.2.5 --- Northern hybridization --- p.31 / Chapter 2.2.6 --- Slot blot hybridization --- p.32 / Chapter 2.2.7 --- Data analysis --- p.33 / Chapter 2.2.8 --- SDS-PAGE analysis --- p.33 / Chapter 2.2.9 --- in situ hybridization --- p.34 / Chapter 2.3 --- Results --- p.37 / Chapter 2.3.1 --- Validation of vitellogenin mRNA detection --- p.37 / Chapter 2.3.2 --- Basal and estradiol-stimulated vitellogenin expression and production invivo --- p.38 / Chapter 2.3.3 --- Localization of vitellogenin expression in the liver --- p.39 / Chapter 2.3.4 --- Expression of vitellogenin in vitro --- p.40 / Chapter 2.4 --- Discussion --- p.54 / Chapter Chapter 3 --- Effects of Steroids on the Expression of Goldfish Vitellogenin in vitro --- p.60 / Chapter 3.1 --- Introduction --- p.60 / Chapter 3.2 --- Materials and Methods --- p.62 / Chapter 3.2.1 --- Materials --- p.62 / Chapter 3.2.2 --- Animal --- p.62 / Chapter 3.2.3 --- Primary culture of dispersed hepatic cells --- p.62 / Chapter 3.2.4 --- Drug treatment --- p.64 / Chapter 3.2.5 --- Total RNA isolation --- p.64 / Chapter 3.2.6 --- Messenger RNA isolation --- p.65 / Chapter 3.2.7 --- Slot blot analysis --- p.66 / Chapter 3.2.8 --- Data analysis --- p.68 / Chapter 3.2.9 --- Reverse transcription-polymerase chain reaction (RT-PCR) --- p.68 / Chapter 3.2.10 --- Cloning of aromatase cDNA --- p.69 / Chapter 3.2.11 --- Sequencing --- p.70 / Chapter 3.3 --- Results --- p.71 / Chapter 3.3.1 --- Effect of 17-β estradiol on vitellogenin mRNA expression --- p.71 / Chapter 3.3.2 --- Effect of testosterone on vitellogenin mRNA expression --- p.71 / Chapter 3.3.3 --- Detection of aromatase mRNA expression in the liver by RT-PCR --- p.72 / Chapter 3.3.4 --- Effect of aromatase inhibitors on testosterone-stimulated vitellogenin expression --- p.73 / Chapter 3.4 --- Discussion --- p.81 / Chapter Chapter 4 --- Effects of Epidermal Growth Factor (EGF) and Activin on the Expression of Vitellogenin in the Goldfish Hepatic Cells in vitro --- p.86 / Chapter 4.1 --- Introduction --- p.86 / Chapter 4.2 --- Materials and Methods --- p.88 / Chapter 4.2.1 --- Materials --- p.88 / Chapter 4.2.2 --- Primary culture of dispersed hepatic cells --- p.89 / Chapter 4.2.3 --- Slot blot analysis --- p.91 / Chapter 4.2.4 --- Data analysis --- p.91 / Chapter 4.3 --- Results --- p.92 / Chapter 4.3.1 --- Effect of activin on vitellogenin mRNA expression --- p.92 / Chapter 4.3.2 --- Effect of EGF and TGF-α on vitellogenin mRNA expression --- p.93 / Chapter 4.4 --- Discussion --- p.99 / Chapter Chapter 5 --- General Discussion --- p.104 / Chapter 5.1 --- Overview --- p.104 / Chapter 5.2 --- Contribution of the present study --- p.106 / Chapter 5.2.1 --- Expression of goldfish vitellogenin in vivo and in vitro --- p.106 / Chapter 5.2.2 --- Effects of steroids on the expression of goldfish vitellogenin in vitro --- p.106 / Chapter 5.2.3 --- Effects of EGF and activin on the expression of vitellogenin in the goldfish hepatic cells in vitro --- p.107 / Chapter 5.3 --- Future prospects --- p.108
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