by Lo Ka-Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 175-197). / Chapter I --- Title page --- p.i / Chapter II --- Thesis committee --- p.ii / Chapter II --- Acknowledgment --- p.iii-iv / Chapter III --- Abstract --- p.v-vi / Chapter IV --- Table of content --- p.vii-xiv / Chapter V --- List of figures --- p.xv-xviii / Chapter VI --- List of tables --- p.xix-xx / Forewords: / Overall objectives --- p.1 / Introduction on the fish used in this research study --- p.2 / Chapter Chapter 1: --- Literature Review on Biomarkers of Stress in Teleosts --- p.5 / Chapter 1.1 --- Definition of stress --- p.7 / Chapter 1.2 --- Classification of stress indicators --- p.8 / Chapter 1.2.1 --- Primary stress indicators --- p.8 / Chapter 1.2.1.1 --- Molecular stress indicators --- p.8 / Chapter 1.2.1.2 --- Hormonal stress indicators --- p.9 / Chapter (I) --- Corticosteroid --- p.9 / Chapter (II) --- Catecholamines --- p.11 / Chapter 1.2.2 --- Secondary stress indicators --- p.12 / Chapter 1.2.2.1 --- Metabolic changes --- p.12 / Chapter (I) --- Glucose metabolism --- p.13 / Chapter (II) --- Lactic acid --- p.14 / Chapter 1.2.2.2 --- Osmoregulatory changes --- p.15 / Chapter 1.2.2.3 --- Haematological changes --- p.16 / Chapter 1.2.2.4 --- Reproductive changes --- p.17 / Chapter 1.2.3 --- Tertiary stress indicators --- p.18 / Chapter 1.2.3.1 --- Histopathological indicators --- p.18 / Chapter 1.2.3.2 --- Ecological indicators --- p.19 / Chapter 1.3 --- Recent trends on the study of biomarkers --- p.20 / Chapter 1.3.1 --- Use of detoxification enzymes for specific indication of toxic pollutants in aquatic environment --- p.20 / Chapter 1.3.1.1 --- Metallothioneins (MTs) --- p.20 / Chapter 1.3.1.2 --- Cytochrome P450 monoxygenase (CYP450) --- p.21 / Chapter 1.3.2 --- Use of HSP 70 as a biomarker in teleost --- p.22 / Chapter 1.4 --- Future perspectives on the study of biomarkers in fish --- p.24 / Chapter Chapter 2: --- Literature Review on Heat Shock Proteins (HSPs) --- p.28 / Chapter 2.1 --- General Characteristics of HSPs --- p.29 / Chapter 2.1.1 --- HSP90 family --- p.30 / Chapter 2.1.2 --- HSP70 family --- p.31 / Chapter 2.1.3 --- HSP60 family (Chaperonin-60) --- p.32 / Chapter 2.1.4 --- Low-molecular weight HSPs (HSP20) --- p.33 / Chapter 2.2 --- Structure of HSP70 encoding gene --- p.33 / Chapter 2.2.1 --- General characteristics of HSP70-encoding gene --- p.33 / Chapter 2.2.2 --- Heat shock transcription factor (HSF) --- p.35 / Chapter 2.2.3 --- Heat shock elements (HSE) --- p.35 / Chapter 2.3 --- Stress-mediated control of HSP70 transcription --- p.36 / Chapter 2.4 --- Characterization of HSP70 expression in teleost --- p.38 / Chapter 2.4.1 --- Tissues-specific expression of HSP70 in teleost --- p.39 / Chapter 2.4.2 --- Inter-relationship of HSP70 expression with seasonal variation and thermotolerance of teleost --- p.40 / Chapter 2.4.3 --- Induction of HSP70 in teleost upon acute thermal stress --- p.41 / Chapter 2.4.4 --- Induction of HSP70 in teleost by non-thermal stressors --- p.43 / Chapter 2.4.4.1 --- Heavy metal-induced HSP70 expression --- p.43 / Chapter 2.4.4.2 --- Handling stress-induced HSP70 expression --- p.43 / Chapter Chapter 3: --- "Induction of HSP70 in blood cells of seabream, Sparus sarba subjected to in vivo and in vitro thermal stress" --- p.48 / Chapter 3.1 --- Introduction --- p.49 / Chapter 3.2 --- Materials and Methods --- p.52 / Chapter 3.2.1 --- Overall experimental design --- p.52 / Chapter 3.2.2 --- Fish --- p.53 / Chapter 3.2.3 --- Blood sampling --- p.53 / Chapter 3.2.4 --- Preparation of blood cells --- p.54 / Chapter 3.2.5 --- Thermal stress regimes --- p.54 / Chapter 3.2.5.1 --- Time couse of HSP70 induction profile in blood cells after in vitro exposure to thermal stress --- p.54 / Chapter 3.2.5.2 --- HSP70 synthesis in blood cells taken from fish subjected to in vivo hyper- thermic stress --- p.55 / Chapter 3.2.5.3 --- Transcriptional inhibitory effect of actinomycin D on the synthesis of HSP70 in blood cells subjected to in vitro thermal stress --- p.55 / Chapter 3.2.6 --- Protein analysis --- p.56 / Chapter 3.2.7 --- Gel electrophoresis --- p.57 / Chapter 3.2.8 --- Immunoblotting (Western blot analysis) --- p.57 / Chapter 3.2.9 --- Autroradiography --- p.58 / Chapter 3.3 --- Results --- p.59 / Chapter 3.3.1 --- Time course of HSP70 induction profile in blood cells subjected to in vitro thermal treatments --- p.59 / Chapter 3.3.1.1 --- Results of immunoblotting from blood cells of fish acclimated to 26°C --- p.59 / Chapter 3.3.1.2 --- Results of immunoblotting in blood cells from 18°C-acclimated fish --- p.60 / Chapter 3.3.1.3 --- Results of immunoblotting in blood cells from fish acclimated to 20°C --- p.60 / Chapter 3.3.1.4 --- 35S-methionine labelling of de novo protein synthesis in blood cells of fish acclimated to 15 and 20°C --- p.61 / Chapter 3.3.2 --- Blood cell HSP70 levels in 20°C-acclimated fish subjected to in vivo hyperthermic stress --- p.61 / Chapter 3.3.3 --- Transcriptional inhibitory effect of actinomycin D on HSP70 induction in blood cells subjected to in vitro thermal stress --- p.62 / Chapter 3.4 --- Discussions --- p.60 / Chapter 3.4.1 --- Characteristics of HSP70 induction in blood cells of seabream subjected to in vitro temperature stress --- p.72 / Chapter 3.4.1.1 --- Induction profile of HSP70 in blood cells --- p.72 / Chapter 3.4.1.2 --- Time course ofHSP70 induction in blood cells --- p.75 / Chapter 3.4.1.3 --- Effect of acclimation temperature of fish on the induction of HSP70 --- p.76 / Chapter 3.4.2 --- Comparison of HSP70 induction in in vitro and in vivo thermal treatment on blood cells --- p.78 / Chapter 3.4.3 --- "Effect of transcriptional inhibitor, actinomycin D, on the de novo synthesis of HSP70" --- p.80 / Chapter 3.5 --- Conclusions --- p.70 / Chapter Chapter 4: --- "Effects of seasonal variation and transportation stress on level of HSP70, serum glucose and serum Cortisol in seabream, Sparus sarba" --- p.86 / Chapter 4.1 --- Introduction --- p.87 / Chapter 4.2 --- Materials and methods --- p.90 / Chapter 4.2.1 --- Overall experimental design --- p.90 / Chapter 4.2.2 --- Fish and blood sampling --- p.91 / Chapter 4.2.3 --- Preparation of blood samples --- p.92 / Chapter 4.2.4 --- Determination of HSP70 levels in blood cells sampled from seabream upon different seasons --- p.92 / Chapter 4.2.5 --- Immunoblotting analysis --- p.92 / Chapter 4.2.6 --- Enzyme-linked Immnosorbent Assay (ELISA) --- p.93 / Chapter 4.2.7 --- Measurement of serum parameter in seabream --- p.95 / Chapter 4.2.7.1 --- Serum glucose --- p.95 / Chapter 4.2.7.2 --- Serum Cortisol --- p.96 / Chapter 4.3 --- Results --- p.96 / Chapter 4.3.1 --- Determination of HSP70 levels in blood cells sampled from seabream in different seasons --- p.96 / Chapter 4.3.1.1 --- Immunoblotting analysis --- p.96 / Chapter 4.3.1.2 --- Enzyme-linked immunosorbent assay (ELISA) --- p.96 / Chapter 4.3.2 --- Serum analysis of seabream sampled from fish farm in different seasons --- p.98 / Chapter 4.3.2.1 --- Serum glucose --- p.98 / Chapter 4.3.2.2 --- Serum Cortisol --- p.99 / Chapter 4.4 --- Discussions --- p.117 / Chapter 4.4.1 --- Characterization of HSP70 expression in blood cells of seabream --- p.117 / Chapter 4.4.2 --- Dynamicity of HSP70 content and thermo- tolerance of fish in different seasons --- p.118 / Chapter 4.4.3 --- Effect of transportation stress on HSP70 induction in blood cells of seabream --- p.121 / Chapter 4.4.4 --- Dynamicity of serum glucose level in seabream subjected to seasonal variations --- p.123 / Chapter 4.4.5 --- Effect of transportation stress on the serum glucose level of seabream in different seasons --- p.124 / Chapter 4.4.6 --- Dynamicity of senam Cortisol level in seabream subjected to seasonal variations --- p.125 / Chapter 4.4.7 --- Effect of transportation stress on the serum Cortisol level of seabream in different seasons --- p.126 / Chapter 4.4.8 --- "Comments on the use of HSP70, serum Cortisol and serum glucose as biomarkersin environmental supervision" --- p.126 / Chapter 4.5 --- Conclusions --- p.129 / Chapter Chapter 5: --- "In vitro and in vivo effects of Cortisol, dexamethasone and adrenaline on the induction of HSP70 in seabream, Sparus sarba" --- p.131 / Chapter 5.1 --- Introduction --- p.132 / Chapter 5.2 --- Materials and methods --- p.133 / Chapter 5.2.1 --- Overall experimental design --- p.133 / Chapter 5.2.2 --- Acclimation of fish and regimes of treatment --- p.133 / Chapter 5.2.3 --- Serum Cortisol and adrenaline analysis --- p.135 / Chapter 5.2.4 --- "Protein analysis, gel electrophoresis, immuno- blotting and ELISA analysis" --- p.136 / Chapter 5.3 --- Results --- p.137 / Chapter 5.3.1 --- "HSP70 level in blood cells treated with Cortisol, dexamethasone and adrenaline in vitro" --- p.137 / Chapter 5.3.2 --- "Serum hormones and HSP70 level in tissues of fish injected with Cortisol, adrenaline and dexamethasone invivo" --- p.137 / Chapter 5.3.2.1 --- Serum Cortisol and adrenaline level of fish after hormone injections --- p.137 / Chapter 5.3.2.2 --- "HSP70 level in blood cells, brain and liver tissue of fish after hormone injections" --- p.138 / Chapter (I) --- Level of HSP70 in blood cells of fish after hormone injections --- p.138 / Chapter 5.4 --- Discussions --- p.156 / Chapter 5.4.1 --- In vitro and in vivo study of the hormonal effect on HSP70 level in blood cells of seabream --- p.156 / Chapter 5.4.2 --- Hypothetical mechanism of hormone-receptor mediated HSP70 regulation --- p.158 / Chapter 5.4.3 --- In vivo study of the hormonal effect on HSP70 level in blood cells of seabream --- p.160 / Chapter 5.4.4 --- In vivo study on the hormonal effect of HSP70 synthesis in liver of seabream --- p.163 / Chapter 5.4.5 --- In vivo study on the hormonal effect of HSP70 synthesis in brain tissue of seabream --- p.165 / Chapter 5.4.6 --- HSP70 level in different tissues of fish in relation to the induction and sensitivity against stress --- p.166 / Chapter 5.5 --- Conclusions --- p.169 / Chapter Chapter 6: --- Summary --- p.171 / References --- p.175
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_321860 |
| Date | January 1997 |
| Contributors | Lo, Ka-Man., Chinese University of Hong Kong Graduate School. Division of Biology. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xx, 197 leaves : ill. (some col.) ; 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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