Sham, Wai Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 173-205). / Abstracts in English and Chinese. / Declaration of Originality --- p.i / Thesis / Assessment Committee --- p.ii / Abstract --- p.iii / 摘要 --- p.vi / Acknowledgements --- p.viii / Table of Contents --- p.x / List of Figures --- p.xiv / List of Tables --- p.xvii / List of Abbreviations --- p.xix / Chapter Chapter I --- Introduction --- p.1 / Chapter 1.1 --- Habitats and diversity of fish --- p.1 / Chapter 1.2 --- Stress accompanied with osmotic changes --- p.2 / Chapter 1.3 --- Physiological consequences associated with osmoregulation --- p.4 / Chapter 1.3.1 --- Energy cost (energy metabolism) --- p.4 / Chapter 1.3.2 --- Food intake --- p.5 / Chapter 1.3.3 --- Reproduction and development --- p.5 / Chapter 1.3.4 --- Growth --- p.6 / Chapter 1.3.5 --- Immunity --- p.6 / Chapter 1.4 --- Osmoregulatory systems in teleosts --- p.7 / Chapter 1.4.1 --- Ion transport --- p.7 / Chapter 1.4.1.1 --- Mechanism in freshwater --- p.8 / Chapter 1.4.1.2 --- Mechanism in seawater --- p.8 / Chapter 1.4.2 --- Water channel --- p.11 / Chapter 1.4.3 --- Accumulation of compatible osmolytes --- p.11 / Chapter 1.5 --- Kinetics on ion transporters --- p.12 / Chapter 1.5.1 --- Na+K+ATPase --- p.12 / Chapter 1.5.2 --- Other ion transporters --- p.14 / Chapter 1.5.3 --- Other factors related to controlling ion transport --- p.15 / Chapter 1.6 --- Pleiotropic hormones involved in osmoregulation --- p.17 / Chapter 1.6.1 --- Growth hormone/ insulin-like factor-1 axis in seawater adaptation --- p.17 / Chapter 1.6.2 --- Prolactin in freshwater adaptation --- p.19 / Chapter 1.6.3 --- "Dual osmoregulatory role of cortisol, the major corticosteroid in fish" --- p.20 / Chapter 1.7 --- "Growth hormone receptor (GHR), prolactin receptor (PRLR) and signaling pathways" --- p.22 / Chapter 1.8 --- Involvement of sex hormones on osmoregulation --- p.25 / Chapter 1.8.1 --- Estrogens --- p.25 / Chapter 1.8.2 --- Testosterone --- p.26 / Chapter 1.9 --- Biological actions of GH through GHR and PRL through PRLR --- p.27 / Chapter 1.9.1 --- Regulation of GHR --- p.27 / Chapter 1.9.2 --- Regulation of PRLR --- p.28 / Chapter 1.9.3 --- Promoter characterization --- p.30 / Chapter 1.10 --- Adaptation to tonicity-ORE/OREBP pathway --- p.32 / Chapter 1.11 --- Animal model --- p.35 / Chapter 1.12 --- Rationale and objectives of the project --- p.35 / Chapter Chapter II --- Materials and methods --- p.38 / Chapter 2.1 --- Animals --- p.38 / Chapter 2.2 --- In vivo experiments --- p.38 / Chapter 2.3 --- In vitro experiments --- p.38 / Chapter 2.4 --- RNA preparation --- p.39 / Chapter 2.5 --- RNA integrity - formaldehyde agarose gel electrophoresis of mRNA --- p.40 / Chapter 2.6 --- Reverse transcription polymerase chain reaction (RT-PCR) --- p.40 / Chapter 2.7 --- Polymerase chain reaction PCR --- p.41 / Chapter 2.8 --- Quantitative Real-time PCR --- p.41 / Chapter 2.9 --- Cell culture --- p.43 / Chapter 2.10 --- Bioinformatics search --- p.43 / Chapter 2.11 --- Plasmid constructions --- p.43 / Chapter 2.12 --- Transient transfection --- p.47 / Chapter 2.13 --- Luciferase assay --- p.47 / Chapter 2.14 --- Site-directed mutagenesis of sbPRLRl and sbPRLR2 gene promoters --- p.48 / Chapter 2.14.1 --- One-round PCR amplification --- p.48 / Chapter 2.14.2 --- Sequential PCR amplification --- p.48 / Chapter 2.15 --- Nuclear extract preparation for electrophoretic mobility shift assays (EMSA) --- p.52 / Chapter 2.16 --- Electrophoretic mobility shift assays (EMSA) --- p.53 / Chapter 2.16.1 --- Preparation of annealed complementary oligonucleotides --- p.53 / Chapter 2.16.2 --- Preparation of radiolabeled oligonucleotides --- p.54 / Chapter 2.16.3 --- Preparation of non-denaturing polyacrylamide gel --- p.54 / Chapter 2.16.4 --- Electrophoretic mobility shift assay --- p.54 / Chapter 2.17 --- Data analysis --- p.56 / Chapter Chapter III --- Result: In vivo gene expression studies --- p.57 / Chapter 3.1 --- Aim of the chapter --- p.57 / Chapter 3.2 --- Results --- p.57 / Chapter 3.2.1 --- Validation of real-time PCR --- p.57 / Chapter 3.2.2 --- "Transcript levels of sbPRLRl, sbPRLR2, sbNKA α- and sbNKA β-subunit after transfer of fish from seawater to hypersaline water" --- p.61 / Chapter 3.2.2.1 --- Gill --- p.61 / Chapter 3.2.2.2 --- Kidney --- p.64 / Chapter 3.3 --- Discussion --- p.67 / Chapter 3.3.1 --- sbPRLR gene expression after seawater-to-hypersaline water transfer --- p.67 / Chapter 3.3.2 --- sbNKA α- and sbNKA β-subunit expression after seawater-to-hypersaline water transfer --- p.71 / Chapter 3.3.3 --- Overall summary --- p.76 / Chapter Chapter IV --- Result: In vitro gene expression studies --- p.77 / Chapter 4.1 --- Aim of the chapter --- p.77 / Chapter 4.2 --- Results --- p.77 / Chapter 4.2.1 --- Gill organ culture --- p.77 / Chapter 4.2.2 --- Kidney explants --- p.81 / Chapter 4.3 --- Discussion --- p.84 / Chapter Chapter V --- Result: Studies on the gene promoter of sbPRLRs under osmotic change --- p.88 / Chapter 5.1 --- Aim of the chapter / Chapter 5.2 --- Results --- p.88 / Chapter 5.2.1 --- The 5´ة-UTR region of the two sbPRLR genes contain several osmotic response elements --- p.88 / Chapter 5.2.2 --- Promoter activities of the 5´ة-flanking regions of the two sbPRLR genes under osmotic stress --- p.91 / Chapter 5.2.3 --- Identification of a DNA region required for the responses to NaCl --- p.97 / Chapter 5.2.4 --- Interaction of the osmotic response region and transcription factors --- p.104 / Chapter 5.2.4.1 --- Identification of DNA-binding activity in nuclear extract --- p.104 / Chapter 5.2.4.2 --- OSTF1 transactivates the sbPRLR gene promoters --- p.107 / Chapter 5.3 --- Discussion --- p.109 / Chapter Chapter VI --- Result: Studies on the gene promoters of sbPRLR under hormonal challenge --- p.125 / Chapter 6.1 --- Aim of the chapter --- p.125 / Chapter 6.2 --- Results --- p.125 / Chapter 6.2.1 --- The 5´ة-UTR region of the two sbPRLR genes contains several hormone response elements --- p.125 / Chapter 6.2.2 --- Identification of a DNA region required for the responses to steroid hormones --- p.126 / Chapter 6.2.2.1 --- E2-dependent regulation of sbPRLR 1 and sbPRLR2 genes use imperfect EREs --- p.126 / Chapter 6.2.2.2 --- An ARE motif in sbPRLR2 gene promoter --- p.132 / Chapter 6.2.2.3 --- An imperfect and a novel GRE present on the sbPRLRl and sbPRLR2 gene promoters respectively --- p.138 / Chapter 6.2.3 --- Interaction of the hormone response region and transcription factors --- p.144 / Chapter 6.2.3.1 --- The ER interacts with the putative ERE motifs in vitro --- p.144 / Chapter 6.2.3.2 --- The AR interacts with the putative ARE motif in vitro --- p.145 / Chapter 6.2.3.3 --- The GR interacts with the putative GRE motifs in vitro --- p.145 / Chapter 6.3 --- Discussion --- p.152 / Chapter 6.3.1. --- Sex steroids --- p.152 / Chapter 6.3.1.1 --- Estrogens --- p.152 / Chapter 6.3.1.2 --- Androgens --- p.157 / Chapter 6.3.2. --- Interrenal hormone ´ؤ cortisol --- p.163 / Chapter Chapter VII --- General conclusion and perspectives --- p.170 / References --- p.173
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326520 |
| Date | January 2008 |
| Contributors | Sham, Wai Yan., Chinese University of Hong Kong Graduate School. Division of Environmental Science. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xx, 205 leaves : ill. ; 30 cm. |
| Rights | Use 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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