Hui Fong Fong Liza. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 130-182). / Abstracts in English and Chinese. / Chapter I --- Title page --- p.I / Chapter II --- Thesis committee --- p.II / Chapter III --- Acknowledgements --- p.III / Chapter IV --- Abstract (Chinese version) --- p.IV / Chapter V --- Abstract (English version) --- p.VII / Chapter VI --- Table of contents --- p.X / Chapter VII --- List of figures --- p.XIV / Chapter VIII --- List of table --- p.XVIII / Chapter Chapter 1: --- General introduction --- p.1 / Chapter Chapter 2: --- Literature review --- p.5 / Chapter 2.1. --- Gill --- p.5 / Chapter 2.2. --- Chloride cells (Mitochondria-rich cells) --- p.6 / Chapter 2.2.1. --- Ion extrusion by fish in seawater --- p.9 / Chapter 2.2.2. --- Ion uptake by fish in hypo-osmotic condition --- p.12 / Chapter 2.3. --- Sparus sarba (Silver seabream) --- p.14 / Chapter 2.4. --- Sodium-potassium adenosinetriphosphatase (Na+-K+-ATPase) --- p.15 / Chapter 2.4.1. --- Na+-K+-ATPase α-subunit --- p.17 / Chapter 2.4.2. --- Na+-K+-ATPase β-subunit --- p.18 / Chapter 2.4.3. --- Regulation of Na+-K+-ATPase --- p.20 / Chapter 2.5. --- Hormones --- p.21 / Chapter 2.5.1. --- Growth hormone-prolactin family --- p.21 / Chapter 2.5.2. --- Structure of hormones --- p.22 / Chapter 2.5.2.1. --- Structure of growth hormone and prolactin in fish --- p.22 / Chapter 2.5.2.2. --- Structure of insulin-like growth factors in fish --- p.26 / Chapter 2.5.2.3. --- Structure of Cortisol in fish --- p.27 / Chapter 2.5.3. --- Regulation of hormones --- p.28 / Chapter 2.5.3.1. --- Regulation of growth hormone in fish --- p.28 / Chapter 2.5.3.2. --- Regulation of prolactin in fish --- p.32 / Chapter 2.5.3.3. --- Regulation of insulin-like growth factor-I in fish --- p.33 / Chapter 2.5.3.4. --- Regulation of Cortisol in fish --- p.33 / Chapter 2.5.4. --- Functions of hormones --- p.33 / Chapter 2.5.4.1. --- Functions of growth hormone in fish --- p.33 / Chapter 2.5.4.2. --- Functions of prolactin in fish --- p.39 / Chapter 2.5.4.3. --- Functions of insulin-like growth factor-I in fish --- p.44 / Chapter 2.5.4.4. --- Functions of Cortisol in fish --- p.45 / Chapter 2.5.4.5. --- "Combined effects of GH, IGF-I, PRL and Cortisol" --- p.49 / Chapter 2.6. --- Salinity effects on Na+-K+-ATPase expression --- p.52 / Chapter Chapter 3: --- In vitro effect of hormones on branchial Na+-K+- ATPase expression in marine teleost Sparus sarba --- p.58 / Chapter 3.1. --- Abstract --- p.58 / Chapter 3.2. --- Introduction --- p.60 / Chapter 3.3. --- Materials and methods --- p.62 / Chapter 3.3.1. --- Overall experimental design --- p.62 / Chapter 3.3.2. --- Fish preparation --- p.62 / Chapter 3.3.3. --- Tissue sampling --- p.62 / Chapter 3.3.4. --- RNA extraction and dot blot analysis --- p.63 / Chapter 3.3.5. --- Protein extraction --- p.65 / Chapter 3.3.6. --- Protein quantification --- p.65 / Chapter 3.3.7. --- Na+-K+-ATPase activity --- p.65 / Chapter 3.3.8. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.66 / Chapter 3.3.9. --- Statistical analysis --- p.67 / Chapter 3.4. --- Results --- p.69 / Chapter 3.4.1. --- Dot blot analysis of Na+-K+-ATPase mRNA subunits --- p.69 / Chapter 3.4.2. --- Analysis of Na+-K+-ATPase protein α-subunit --- p.81 / Chapter 3.4.3. --- Analysis of Na+-K+-ATPase activity --- p.87 / Chapter 3.5. --- Discussion --- p.92 / Chapter 3.5.1. --- Effects of rbGH and rbIGF-I on Na+-K+-ATPase expression --- p.92 / Chapter 3.5.2. --- Effects of oPRL on Na+-K+-ATPase expression --- p.102 / Chapter 3.5.3 --- Effects of Cortisol on Na+-K+-ATPase expression --- p.104 / Chapter 3.6. --- Conclusion --- p.108 / Chapter Chapter 4: --- In vivo effect of salinity on branchial Na+-K+-ATPase expression in marine teleost Sparus sarba --- p.109 / Chapter 4.1. --- Abstract --- p.109 / Chapter 4.2. --- Introduction --- p.110 / Chapter 4.3. --- Materials and methods --- p.112 / Chapter 4.3.1. --- Overall experimental design --- p.112 / Chapter 4.3.2. --- Fish preparation --- p.112 / Chapter 4.3.3. --- Tissue sampling --- p.113 / Chapter 4.3.4. --- "RNA extraction, dot blot analysis, protein extraction, quantification, Na+-K+-ATPase activity, protein gel electrophoresis and immunoblotting (Western blotting)" --- p.113 / Chapter 4.3.5. --- Statistical analysis --- p.114 / Chapter 4.4. --- Results --- p.114 / Chapter 4.4.1. --- Dot blot analysis of Na+-K+-ATPase mRNA subunits --- p.114 / Chapter 4.4.2. --- Analysis of Na+-K+-ATPase protein a-subunit --- p.114 / Chapter 4.4.3. --- Analysis of Na+-K+-ATPase activity --- p.115 / Chapter 4.5. --- Discussion --- p.120 / Chapter 4.6. --- Conclusion --- p.125 / Chapter Chapter 5: --- General discussion and conclusion --- p.126 / References --- p.130

Ng Ho Yuen Andus. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 105-131). / Abstracts in English and Chinese. / Chapter I --- Title page --- p.i / Chapter II --- Thesis committee --- p.ii / Chapter III --- Acknowledgement --- p.iii / Chapter IV --- Abstract --- p.v / Chapter V --- Abstract (Chinese version) --- p.vii / Chapter V --- Table of contents --- p.ix / Chapter VI --- List of abbreviations --- p.xv / Chapter VII --- List of figures --- p.xviii / General introduction --- p.1 / Chapter Chapter 1: --- Literature review --- p.5 / Chapter 1.1. --- Heat shock proteins (HSPs) --- p.6 / Chapter 1.1.1. --- Introduction --- p.6 / Chapter 1.1.2. --- The various heat shock proteins --- p.8 / Chapter 1.1.2.1. --- HSP100s --- p.8 / Chapter 1.1.2.2. --- HSP90s --- p.9 / Chapter 1.1.2.3. --- HSP70s --- p.12 / Chapter 1.1.2.3.1. --- ATPase reaction cycle of HSP70 and protein folding --- p.13 / Chapter 1.1.2.3.2. --- Protein translocation --- p.14 / Chapter 1.1.2.3.3. --- Selective lysosomal proteolysis --- p.16 / Chapter 1.1.2.4. --- HSP60s --- p.16 / Chapter 1.1.2.5. --- Small HSPs --- p.17 / Chapter 1.1.2.6. --- Ubiquitin --- p.19 / Chapter 1.1.3. --- HSP studies in fish --- p.21 / Chapter 1.1.3.1. --- In vivo works --- p.21 / Chapter 1.1.3.2. --- In vitro works --- p.23 / Chapter 1.2. --- Growth hormone / prolactin family in teleostean fishes --- p.26 / Chapter 1.2.1. --- Introduction --- p.26 / Chapter 1.2.2. --- Growth hormone (GH; somatotropin) --- p.29 / Chapter 1.2.2.1. --- Structure --- p.29 / Chapter 1.2.2.2. --- Actions --- p.29 / Chapter 1.2.2.3. --- Insulin-like Growth Factors (IGFs; somatomedins) --- p.31 / Chapter 1.2.3. --- Prolactin (PRL) --- p.34 / Chapter 1.2.3.1. --- Structure --- p.34 / Chapter 1.2.3.2. --- Actions --- p.35 / Chapter 1.2.4. --- Somatolactin (SL) --- p.37 / Chapter 1.2.4.1. --- Structure --- p.37 / Chapter 1.2.4.2. --- Actions --- p.38 / Chapter 1.2.5. --- Growth hormone receptor (GH-R) and prolactin receptor (PRL-R) --- p.39 / Chapter 1.3. --- Cortisol in teleostean fishes --- p.41 / Chapter 1.4. --- Salinity adaptation in teleosts --- p.44 / Chapter Chapter 2: --- Effect of in vitro thermal shock on HSP70 expression in whole blood of Sparus sarba --- p.46 / Chapter 2.1. --- Introduction --- p.47 / Chapter 2.2. --- Materials and methods --- p.49 / Chapter 2.2.1. --- Overall experimental design --- p.49 / Chapter 2.2.2. --- Experimental fish --- p.49 / Chapter 2.2.3. --- Blood sampling and preparation --- p.49 / Chapter 2.2.4. --- Thermal stress regimes --- p.50 / Chapter 2.2.5. --- Protein extraction --- p.51 / Chapter 2.2.6. --- Protein quantification --- p.51 / Chapter 2.2.7. --- Indirect enzyme-linked immunosorbent assay (ELISA) --- p.52 / Chapter 2.2.8. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.54 / Chapter 2.2.9. --- Statistical analysis --- p.55 / Chapter 2.3. --- Results --- p.56 / Chapter 2.3.1. --- Validation of indirect ELISA --- p.56 / Chapter 2.3.2. --- Effect of in vitro thermal shock on HSP70 expression in whole blood of Sparus sarba --- p.56 / Chapter 2.4. --- Discussion --- p.60 / Chapter 2.5. --- Conclusion --- p.64 / Chapter Chapter 3: --- Effects of hormones on HSP70 expression in whole blood of Sparus sarba in vitro --- p.65 / Chapter 3.1. --- Introduction --- p.66 / Chapter 3.2. --- Materials and methods --- p.68 / Chapter 3.2.1. --- Overall experimental design and experimental fish --- p.68 / Chapter 3.2.2. --- Hormone treatments --- p.59 / Chapter 3.2.3. --- "Protein extraction and quantification, indirect ELISA，gel electrophoresis, and immunoblotting (Western blotting)" --- p.70 / Chapter 3.2.4. --- Statistical analysis --- p.70 / Chapter 3.3. --- Results --- p.71 / Chapter 3.3.1. --- Effect of Cortisol on HSP70 levels in whole Blood --- p.71 / Chapter 3.3.2. --- Effect of recombinant bream growth hormone on HSP70 levels in whole blood --- p.71 / Chapter 3.3.3. --- Effect of recombinant bream insulin-like growth factor-I on HSP70 levels in whole blood --- p.71 / Chapter 3.3.4. --- Effect of ovine prolactin on HSP70 levels in whole blood --- p.72 / Chapter 3.4. --- Discussion --- p.81 / Chapter 3.4.1. --- Effect of Cortisol on HSP70 levels in whole Blood --- p.81 / Chapter 3.4.2. --- Effect of recombinant bream growth hormone on HSP70 levels in whole blood --- p.83 / Chapter 3.4.3. --- Effect of recombinant bream insulin-like growth factor-I on HSP70 levels in whole blood --- p.85 / Chapter 3.4.4. --- Effect of ovine prolactin on HSP70 levels in whole blood --- p.86 / Chapter 3.5. --- Conclusion --- p.88 / Chapter Chapter 4: --- Effect on HSP70 expression in whole blood of Sparus sarba acclimated to various salinities --- p.89 / Chapter 4.1. --- Introduction --- p.90 / Chapter 4.2. --- Materials and methods --- p.92 / Chapter 4.2.1. --- Overall experimental design and experimental fish --- p.92 / Chapter 4.2.2. --- "Protein extraction and quantification, indirect ELISA, gel electrophoresis, and immunoblotting (Western blotting)" --- p.92 / Chapter 4.2.3. --- Statistical analysis --- p.93 / Chapter 4.3. --- Results --- p.94 / Chapter 4.4. --- Discussion --- p.97 / Chapter 4.5. --- Conclusion --- p.100 / Chapter Chapter 5: --- General discussion and conclusion --- p.101 / References --- p.105