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Effects of cadmium exposure on hormonal status and expression of metallothionein and glucose-6-phosphate dehydrogenase in silver sea bream, Sparus sarba.

Man, Ka Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 101-126). / 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 (English) --- p.iv / Chapter V. --- Abstract (Chinese) --- p.vi / Chapter VI. --- Table of contents --- p.vii / Chapter VII. --- List of abbreviations --- p.xiii / Chapter VIII. --- List of figures --- p.xv / General introduction --- p.1 / Chapter Chapter 1: --- Literature review --- p.4 / Chapter 1.1. --- Cadmium --- p.5 / Chapter 1.1.1. --- Cadmium - Ways of uptake in human and aquatic life --- p.5 / Chapter 1.1.2. --- Cadmium - Toxic effects in fish --- p.6 / Chapter 1.2. --- Cortisol --- p.11 / Chapter 1.2.1. --- Cortisol - General information and its regulations --- p.11 / Chapter 1.2.2. --- Cortisol - Functions --- p.12 / Chapter 1.3. --- Thyroid hormones --- p.14 / Chapter 1.3.1. --- THs - General information and its regulations --- p.14 / Chapter 1.3.2. --- THs - Functions --- p.15 / Chapter 1.4. --- Growth hormone --- p.18 / Chapter 1.4.1. --- GH - General information and its regulations --- p.18 / Chapter 1.4.2. --- GH - Functions --- p.20 / Chapter 1.5. --- Insulin-like growth factor --- p.22 / Chapter 1.5.1. --- IGF-I - General information and its regulations --- p.22 / Chapter 1.5.2. --- IGF-I - Functions --- p.24 / Chapter 1.6 --- Metallothioneins --- p.26 / Chapter 1.6.1. --- MTs - Definition and Classification --- p.26 / Chapter 1.6.2. --- MTs - Functions --- p.27 / Chapter 1.7. --- Glucose-6-phosphate dehydrogenase --- p.31 / Chapter 1.7.1. --- G6PDH - General information and its regulations --- p.31 / Chapter 1.7.2. --- G6PDH ´ؤ Functions --- p.32 / Chapter Chapter 2: --- "Effects of cadmium exposure on the endocrine status of silver sea bream, Sparus sarba" --- p.34 / Chapter 2.1. --- Introduction --- p.35 / Chapter 2.2. --- Materials and methods --- p.37 / Chapter 2.2.1. --- Overall experimental design --- p.37 / Chapter 2.2.2. --- In vivo exposure to waterborne cadmium --- p.37 / Chapter 2.2.2.1. --- Experimental animals --- p.37 / Chapter 2.2.2.2. --- Adaptation --- p.37 / Chapter 2.2.2.3. --- Tissue sampling --- p.38 / Chapter 2.2.2.4. --- Enzyme-linked immunosorbent assay (ELISA) --- p.38 / Chapter 2.2.2.4.1. --- Serum cortisol analysis --- p.39 / Chapter 2.2.2.4.2. --- Serum triiodothyronine analysis --- p.39 / Chapter 2.2.2.4.3. --- Serum thyroxine analysis --- p.39 / Chapter 2.2.2.5. --- Protein extraction and Protein quantification --- p.40 / Chapter 2.2.2.6. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.40 / Chapter 2.2.2.7. --- RNA extraction --- p.41 / Chapter 2.2.2.8. --- Reverse transcription for first-strand cDNAs from total RNAs samples from liver --- p.42 / Chapter 2.2.2.9. --- Real-time quantitative PCR assays of IGF-I mRNA expression --- p.43 / Chapter 2.2.3. --- In vivo experiments involving cadmium injection --- p.44 / Chapter 2.2.3.1. --- Experimental animals --- p.44 / Chapter 2.2.3.2. --- Adaptation --- p.44 / Chapter 2.2.3.3. --- Tissue sampling --- p.45 / Chapter 2.2.3.4. --- Enzyme-linked immunosorbent assay (ELISA) --- p.45 / Chapter 2.2.3.4.1. --- Serum cortisol analysis --- p.45 / Chapter 2.2.3.4.2. --- Serum triiodothyronine analysis --- p.45 / Chapter 2.2.3.4.3. --- Serum thyroxine analysis --- p.46 / Chapter 2.2.3.5. --- Protein extraction and Protein quantification --- p.46 / Chapter 2.2.3.6. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.46 / Chapter 2.2.3.7. --- RNA extraction --- p.46 / Chapter 2.2.3.8. --- Reverse transcription for the first-strand cDNAs from total RNAs samples from liver --- p.46 / Chapter 2.2.3.9. --- Real-time quantitative PCR of IGF-I mRNA expression --- p.46 / Chapter 2.2.4. --- In vitro part of the project (Primary cell culture: hepatocytes exposed to cadmium medium) --- p.47 / Chapter 2.2.4.1. --- Experimental animals --- p.47 / Chapter 2.2.4.2. --- Primary hepatocytes culture preparation --- p.47 / Chapter 2.2.4.3. --- Cadmium treatment and cell harvest --- p.48 / Chapter 2.2.4.4. --- "RNA extraction, reverse transcription for the first-strand cDNAs from total RNAs samples from lysed cells and real-time quantitative PCR of IGF-I mRNA expression" --- p.48 / Chapter 2.2.5. --- Statistical analysis --- p.48 / Chapter 2.3. --- Results --- p.49 / Chapter 2.3.1. --- Serum cortisol level --- p.49 / Chapter 2.3.2. --- Serum triiodothyronine level --- p.49 / Chapter 2.3.3. --- Serum thyroxine level --- p.49 / Chapter 2.3.4. --- Pituitary growth hormone level --- p.50 / Chapter 2.3.5. --- Hepatic insulin-like growth factor mRNA expression --- p.50 / Chapter 2.4. --- Discussion --- p.58 / Chapter 2.4.1. --- Serum cortisol level --- p.58 / Chapter 2.4.2. --- Thyroid hormones --- p.61 / Chapter 2.4.3. --- Growth hormone --- p.64 / Chapter 2.4.4. --- Insulin-like growth factor-I --- p.67 / Chapter 2.5. --- Conclusion --- p.70 / Chapter Chapter 3: --- "Effects of cadmium exposure on MT and G6PDH mRNA expressions of silver sea bream, Sparus sarba" --- p.71 / Chapter 3.1. --- Introduction --- p.72 / Chapter 3.2. --- Materials and methods --- p.74 / Chapter 3.2.1. --- Overall experimental design --- p.74 / Chapter 3.2.2. --- In vivo experiments involving exposure to waterborne cadmium --- p.74 / Chapter 3.2.2.1. --- Experimental animals --- p.74 / Chapter 3.2.2.2. --- Adaptation --- p.74 / Chapter 3.2.2.3. --- Tissue sampling --- p.74 / Chapter 3.2.2.4. --- RNA extraction --- p.74 / Chapter 3.2.2.5. --- Reverse transcription for first-strand cDNAs from total RNAs samples from gill and liver --- p.75 / Chapter 3.2.2.6. --- Amplification of partial fragments of metallothionein (MT) --- p.75 / Chapter 3.2.2.7. --- Rapid amplification of 5´ة and 3´ة cDNA ends of metallothionein (MT) --- p.76 / Chapter 3.2.2.7.1. --- Amplification of 5´ة cDNA end --- p.76 / Chapter 3.2.2.7.2. --- Amplification of 3´ة cDNA end --- p.78 / Chapter 3.2.2.8. --- Real-time quantitative PCR of MT and G6PDH mRNA expressions --- p.79 / Chapter 3.2.3. --- In vivo injection of cadmium --- p.80 / Chapter 3.2.3.1. --- "Experimental animals, adaptation and tissue sampling" --- p.80 / Chapter 3.2.3.2. --- RNA extraction --- p.80 / Chapter 3.2.3.3. --- Reverse transcription for first-strand cDNAs from total RNAs samples from gill and liver --- p.81 / Chapter 3.2.3.4. --- Real-time quantitative PCR of MT and G6PDH mRNA expression --- p.81 / Chapter 3.2.4. --- In vitro exposure of primary hepatocyte culture to cadmium --- p.81 / Chapter 3.2.4.1. --- Experimental animals --- p.81 / Chapter 3.2.4.2. --- Preparation of the hepatocytes for cell culture --- p.81 / Chapter 3.2.4.3. --- Cadmium treatment and cell harvest --- p.81 / Chapter 3.2.4.4. --- "RNA extraction, reverse transcription for first-strand cDNAs from total RNAs samples from lysed cells and real-time quantitative PCR of MT and G6PDH mRNA expression" --- p.82 / Chapter 3.2.5. --- Statistical analysis --- p.82 / Chapter 3.3. --- Results --- p.83 / Chapter 3.3.1. --- MT cloning and characterization --- p.83 / Chapter 3.3.2. --- Metallothionein mRNA expression --- p.83 / Chapter 3.3.3. --- Hepatic glucose-6-phosphate dehydrogenase mRNA expression --- p.84 / Chapter 3.4. --- Discussion --- p.91 / Chapter 3.4.1. --- MT cloning and characterization --- p.91 / Chapter 3.4.2. --- Metallothioneins mRNA expression --- p.92 / Chapter 3.4.3. --- Hepatic glucose-6-phosphate dehydrogenase mRNA expression --- p.95 / Chapter 3.5. --- Conclusion --- p.98 / General conclusion --- p.99 / References --- p.101

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326214
Date January 2008
ContributorsMan, Ka Yan., Chinese University of Hong Kong Graduate School. Division of Biology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xvii, 126 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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