Szeto Tsz Kwan Leo. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 112-120). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Tables --- p.ix / List of Figures --- p.x / Abbreviations --- p.xii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Heavy metals contaminations in Shing Mun River --- p.1 / Chapter 1.1 --- Importance of copper regulation and role of liverin copper metabolism --- p.6 / Chapter 1.1.1 --- Role of copper --- p.6 / Chapter 1.1.2 --- Toxicity due to unbalanced copper regulation --- p.7 / Chapter 1.1.3 --- Function of liver in copper detoxification --- p.9 / Chapter 1.2 --- Aims and rationale of this research --- p.11 / Chapter Chapter 2 --- Heavy metal concentrations of tilapia samples collected from Shing Mun River --- p.12 / Chapter 2.1 --- Introduction --- p.12 / Chapter 2.1.1 --- Sampling sites - Fo Tan and Siu Lek Yuen Nullah --- p.12 / Chapter 2.1.2 --- Tilapia samples collected from the sites --- p.16 / Chapter 2.1.3 --- Tilapia as a study model --- p.18 / Chapter 2.1.4 --- Bioavailability of heavy metals in water --- p.19 / Chapter 2.1.5 --- Metal content in liver --- p.20 / Chapter 2.1.6 --- Aim of this chapter --- p.20 / Chapter 2.2 --- Materials and Methods --- p.22 / Chapter 2.2.1 --- Collection of control and field samples --- p.22 / Chapter 2.2.2 --- Heavy metal concentrations determination --- p.23 / Chapter 2.2.3 --- Homogenization of liver cells --- p.24 / Chapter 2.2.4 --- Subcellular fractionation --- p.24 / Chapter 2.2.5 --- Determination of copper and zinc content in each subcellular fraction --- p.253 / Chapter 2.3 --- Results --- p.27 / Chapter 2.3.1 --- Physical data --- p.27 / Chapter 2.3.2 --- Metal concentrations in liver and muscle --- p.27 / Chapter 2.3.3 --- Copper and zinc subcellular distribution in liver cell --- p.33 / Chapter 2.4 --- Discussion --- p.36 / Chapter 2.4.1 --- Difference in metal concentration between sites --- p.36 / Chapter 2.4.2 --- Copper contamination in water and fish organ (muscle and liver) from the Shing Mun River --- p.36 / Chapter 2.4.3 --- Comparison of metal content in muscle and liver at Fo Tan site with previous studies --- p.39 / Chapter 2.4.4 --- Copper and zinc concentrations in the liver of tilapia --- p.42 / Chapter 2.4.5 --- Copper and zinc sebcellular distribution in the liver of tilapia --- p.43 / Chapter Chapter 3 --- Column chromatography of hepatic proteins from tilapias --- p.44 / Chapter 3.1 --- Transport of metals from circulatory system to liver --- p.44 / Chapter 3.1.1 --- Copper transporting plasma proteins in vertebrates --- p.44 / Chapter 3.1.2 --- Copper uptake into hepatocytes --- p.45 / Chapter 3.1.3 --- Intracellular metabolism of copper --- p.48 / Chapter 3.1.4 --- Mechanism of copper toxicity following excess accumulation --- p.49 / Chapter 3.1.5 --- Aim of this chapter --- p.50 / Chapter 3.2 --- Materials and Methods --- p.51 / Chapter 3.2.1 --- Purification of liver cytosolic proteins by gel-filtration column chromatography --- p.51 / Chapter 3.2.2 --- Copper content detection in elution --- p.52 / Chapter 3.2.3 --- Analysis of peaks from elution profile using tricine gel SDS PAGE --- p.53 / Chapter 3.3 --- Results --- p.55 / Chapter 3.3.1 --- Gel-filtration liquid chromatography elution profiles --- p.55 / Chapter 3.3.2 --- SDS PAGE analysis of peaks in elution profiles --- p.51 / Chapter 3.4 --- Discussion --- p.54 / Chapter 3.4.1 --- Comparison of gel filtration profiles of sample liver cytosol between sites and sexes --- p.64 / Chapter 3.4.2 --- Possible proteins in peaks found in the gel filtration profiles --- p.64 / Chapter 3.4.3 --- Common copper-indeced proteins --- p.67 / Chapter 3.5 --- Conclusion --- p.70 / Chapter Chapter 4 --- Two-dimensional electrophoresis of hepatic cutosol of tilapias caught from Shing Mun River and copper-treated HEPA T1 cell --- p.72 / Chapter 4.1 --- Introduction --- p.72 / Chapter 4.1.1 --- The need of ´بin vitro' experiment --- p.72 / Chapter 4.1.2 --- Choice of cell line --- p.73 / Chapter 4.1.3 --- Aim of this chapter --- p.74 / Chapter 4.2 --- Materials and Methods --- p.76 / Chapter 4.2.1 --- HEPA T1 cell cultivation --- p.76 / Chapter 4.2.2 --- Copper exposure of HEPA T1 cell --- p.77 / Chapter 4.2.3 --- Subcellular protein extraction of the copper-treated HEPA T1 cells --- p.77 / Chapter 4.2.4 --- Bicinchoninic Acidic (BCA) Protein Assay --- p.79 / Chapter 4.2.5 --- Two-dimensional gel electrophoresis --- p.79 / Chapter 4.3 --- Results --- p.83 / Chapter 4.3.1 --- Graphical presentation of spots observed on 2-dimensional gel of field samples and copper-injected samples --- p.33 / Chapter 4.3.2 --- Graphical presentation of spots detected on 2-dimensional gel of HEPAT1 cells --- p.84 / Chapter 4.3.3 --- Comparison of matched spots on 2-dimensional gels among control and copper-treated HEPAT1 cells --- p.97 / Chapter 4.4 --- Discussion --- p.105 / Chapter 4.4.1 --- Comparison of the spot patterns between field sample and copperOtreated HEPA T1 cells --- p.105 / Chapter 4.5 --- Conclusion --- p.107 / Chapter Chapter 5 --- General Discussions --- p.108 / Chapter 5.2 --- Research Overview --- p.108 / Chapter 5.2 --- Characterization of metal binding proteins from the cytosol of liver of tilapia --- p.109 / REFERENCES --- p.112