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Cloning and characterization of follistatin in the goldfish, Carassius auratus.

Cheng Fu Yip Gheorghe. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 97-116). / Abstracts in English and Chinese. / Acknowledgement --- p.I / Abstract (in English) --- p.III / Abstract (in Chinese) --- p.V / Table of Content --- p.VII / Symbols and Abbreviations --- p.XII / Scientific Names --- p.XIV / List of Tables --- p.XV / List of Figures --- p.XVI / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Gonadotropin / Chapter 1.1.1 --- Structure --- p.2 / Chapter 1.1.2 --- Function --- p.3 / Chapter 1.1.3 --- Regulation --- p.4 / Chapter 1.1.3.1 --- Neuroendocrine and endocrine regulation of GTHs --- p.4 / Chapter 1.1.3.1.1 --- Hypothalamic neuropeptides and neurotransmitters --- p.6 / Chapter 1.1.3.1.2 --- Gonadal steroids --- p.7 / Chapter 1.1.3.2 --- Paracrine regulation of GTH --- p.8 / Chapter 1.2 --- Activin / Chapter 1.2.1 --- Structure --- p.8 / Chapter 1.2.2 --- Function --- p.9 / Chapter 1.2.3 --- Regulation of activin activity --- p.12 / Chapter 1.2.3.1 --- Intracellular blockade of activin signaling by Smad7 --- p.12 / Chapter 1.2.3.2 --- Extracellular control of activin access --- p.13 / Chapter 1.2.3.2.1 --- Inhibin --- p.13 / Chapter 1.2.3.2.2 --- Activin-binding protein --- p.14 / Chapter 1.3 --- Follistatin / Chapter 1.3.1 --- Structure --- p.14 / Chapter 1.3.2 --- Function --- p.16 / Chapter 1.3.3 --- Regulation in the pituitary --- p.19 / Chapter 1.4 --- Objectives of the Present Study --- p.20 / Chapter Chapter 2 --- Cloning and Recombinant Production of Goldfish Follistatin / Chapter 2.1 --- Introduction --- p.24 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Reagents --- p.26 / Chapter 2.2.2 --- Animal --- p.26 / Chapter 2.2.3 --- Extraction of total RNA and reverse transcription --- p.27 / Chapter 2.2.4 --- Cloning of full-length cDNA encoding goldfish follistatin --- p.27 / Chapter 2.2.5 --- Sequencing of the cDNA --- p.29 / Chapter 2.2.6 --- Distribution of follistatin mRNA in different tissues --- p.29 / Chapter 2.2.7 --- Production of rgFS --- p.30 / Chapter 2.2.8 --- RT-PCR of the rgFS-positive clones --- p.34 / Chapter 2.2.9 --- Extraction of genomic DNA from rgFS-positive clones --- p.34 / Chapter 2.2.10 --- Functional analysis of rgFS --- p.35 / Chapter 2.2.11 --- Data Analysis --- p.37 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Cloning and sequence analysis of goldfish follistatin --- p.37 / Chapter 2.3.2 --- Tissue distribution of follistatin mRNA in the goldfish --- p.39 / Chapter 2.3.3 --- Production and bioassay of rgFS --- p.43 / Chapter 2.4 --- Discussion --- p.47 / Chapter Chapter 3 --- Function and Regulation of Follistatin in the Goldfish Pituitary; Evidence for an Intrinsic Activin/Follistatin Regulatory Feedback Loop / Chapter 3.1 --- Introduction --- p.54 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Reagents --- p.57 / Chapter 3.2.2 --- Animals --- p.57 / Chapter 3.2.3 --- Primary culture of dispersed pituitary cells --- p.57 / Chapter 3.2.4 --- RNA extraction and reverse transcription --- p.58 / Chapter 3.2.5 --- Ovariectomy on pituitary follistatin expression --- p.5 9 / Chapter 3.2.6 --- Seasonal expression profile of follistatin --- p.59 / Chapter 3.2.7 --- Validation of semi-quantitative RT-PCR assays --- p.61 / Chapter 3.2.8 --- Real-time PCR for assay on follistatin and β-actin expression --- p.61 / Chapter 3.2.9 --- Data analysis --- p.63 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Expression of follistatin in the goldfish pituitary --- p.64 / Chapter 3.3.2 --- Validation of semi-quantitative RT-PCR assay --- p.64 / Chapter 3.3.3 --- Activin regulation of pituitary follistatin --- p.64 / Chapter 3.3.4 --- Effects of sex steroids on pituitary follistatin expression --- p.69 / Chapter 3.3.5 --- Effect of GnRH on follistatin expression in the pituitary --- p.74 / Chapter 3.3.6 --- Effect of intracellular cAMP level on pituitary follistatin expression --- p.74 / Chapter 3.3.7 --- Seasonal variation profile of goldfish pituitary follistatin --- p.78 / Chapter 3.4 --- Discussion --- p.78 / Chapter Chapter 4 --- General Discussion / Chapter 4.1 --- Overview --- p.89 / Chapter 4.2 --- Contribution of the Present Study / Chapter 4.2.1 --- Cloning of full-length goldfish follistatin cDNA --- p.91 / Chapter 4.2.2 --- Establishment of stable cell line for expression of rgFS --- p.92 / Chapter 4.2.3 --- Evidence for the presence of intrinsic feedback loop of activin in the goldfish pituitary --- p.92 / Chapter 4.2.4 --- Modulation of follistatin expression in the pituitary by sex steroids --- p.93 / Chapter 4.2.5 --- Conclusions --- p.93 / Chapter 4.3 --- Future Prospects / Chapter 4.3.1 --- Production of rgFS --- p.95 / Chapter 4.3.2 --- Regulation of activin-follistatin system in the pituitary --- p.95 / Reference --- p.96

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324246
Date January 2003
ContributorsCheng, Fu Yip Gheorghe., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xviii, 116 leaves : ill. ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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