Lee Wing Sum. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 206-226). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract (Chinese Version) --- p.iv / Acknowledgements --- p.vii / Table of Contents --- p.viii / List of Abbreviations --- p.xiv / List of Figures --- p.xvii / List of Tables --- p.xxiv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- Literature review --- p.3 / Chapter 2.1 --- Peroxisomes --- p.3 / Chapter 2.2 --- Peroxisome proliferators --- p.5 / Chapter 2.3 --- Human exposure pathways to peroxisome proliferators --- p.5 / Chapter 2.4 --- Peroxisome proliferator-induced pleiotropic effects in rodents --- p.7 / Chapter 2.4.1 --- Short-term effects --- p.7 / Chapter 2.4.1.1 --- Hepatomegaly --- p.7 / Chapter 2.4.2.1 --- Peroxisome proliferation --- p.8 / Chapter 2.4.1.3 --- Alteration of gene transcriptions --- p.8 / Chapter 2.4.2 --- Long-term effect --- p.9 / Chapter 2.5 --- Mechanisms of actions of peroxisome proliferators --- p.9 / Chapter 2.5.1 --- Substrate overload --- p.9 / Chapter 2.5.2 --- Receptor-mediated --- p.11 / Chapter 2.6 --- Peroxisome proliferator-activated receptors (PPARs) --- p.11 / Chapter 2.6.1 --- Structure of PPARs --- p.11 / Chapter 2.6.2 --- Tissue-specific expression of PPARs --- p.15 / Chapter 2.6.3 --- Physiological functions of PPARs --- p.19 / Chapter 2.6.3.1 --- PPARα --- p.19 / Chapter 2.6.3.2 --- PPARγ --- p.21 / Chapter 2.6.3.3 --- PPARδ --- p.23 / Chapter 2.7 --- Role of PPARα involved in peroxisome proliferator-induced pleiotropic responses --- p.24 / Chapter 2.7.1 --- Short-term effects --- p.24 / Chapter 2.7.2 --- Long-term effect --- p.24 / Chapter 2.8 --- Mechanisms of peroxisome proliferator-induced hepatocarcinogenesis --- p.25 / Chapter 2.8.1 --- Oxidative stress --- p.25 / Chapter 2.8.2 --- Suppression of apoptosis --- p.26 / Chapter 2.8.3 --- Increased cell proliferation --- p.27 / Chapter 2.9 --- Species difference to peroxisome proliferator-induced pleiotropic effects --- p.28 / Chapter 2.10 --- Fluorescent differential display (FDD) --- p.32 / Chapter Chapter 3 --- Objectives --- p.35 / Chapter Chapter 4 --- Materials and methods --- p.37 / Chapter 4.1 --- Animals and treatments --- p.37 / Chapter 4.1.1 --- Materials --- p.37 / Chapter 4.1.2 --- Methods --- p.37 / Chapter 4.2 --- Serum triglyceride and cholesterol analyses --- p.39 / Chapter 4.2.1 --- Materials --- p.41 / Chapter 4.2.2 --- Methods --- p.41 / Chapter 4.2.2.1 --- Serum preparation --- p.41 / Chapter 4.2.2.2 --- Triglyceride determination --- p.41 / Chapter 4.2.2.3 --- Cholesterol determination --- p.42 / Chapter 4.3 --- Statistical analysis --- p.42 / Chapter 4.4 --- Tail-genotyping --- p.42 / Chapter 4.4.1 --- Materials --- p.44 / Chapter 4.4.2 --- Methods. --- p.44 / Chapter 4.4.2.1 --- Preparation of genomic tail DNA --- p.44 / Chapter 4.4.2.2 --- PCR reaction --- p.45 / Chapter 4.5 --- Total RNA isolation --- p.45 / Chapter 4.5.1 --- Materials --- p.48 / Chapter 4.5.2 --- Methods --- p.48 / Chapter 4.6 --- DNase I treatment --- p.48 / Chapter 4.6.1 --- Materials --- p.49 / Chapter 4.6.2 --- Methods --- p.49 / Chapter 4.7 --- Reverse transcription of mRNA and fluorescent PCR amplification --- p.50 / Chapter 4.7.1 --- Materials --- p.50 / Chapter 4.7.2 --- Methods --- p.53 / Chapter 4.8 --- Fluorescent differential display (FDD) --- p.53 / Chapter 4.8.1 --- Materials --- p.53 / Chapter 4.8.2 --- Methods --- p.54 / Chapter 4.9 --- Excision of differentially expressed cDNA fragments --- p.54 / Chapter 4.9.1 --- Materials --- p.57 / Chapter 4.9.2 --- Methods --- p.57 / Chapter 4.10 --- Reamplification of differentially expressed fragments --- p.57 / Chapter 4.10.1 --- Materials --- p.60 / Chapter 4.10.2 --- Methods --- p.60 / Chapter 4.11 --- Subcloning of reamplified cDNA fragments --- p.62 / Chapter 4.11.1 --- PCR-TRAP® cloning system --- p.62 / Chapter 4.11.1.1 --- Materials --- p.63 / Chapter 4.11.1.2 --- Methods --- p.63 / Chapter 4.11.2 --- AdvaTage´ёØ PCR cloning system --- p.65 / Chapter 4.11.2.1 --- Materials --- p.65 / Chapter 4.11.2.2 --- Methods --- p.66 / Chapter 4.12 --- Purification of plasmid DNA from recombinant clones --- p.69 / Chapter 4.12.1 --- Materials --- p.69 / Chapter 4.12.2 --- Methods --- p.69 / Chapter 4.13 --- DNA sequencing of differentially expressed cDNA fragments --- p.70 / Chapter 4.13.1 --- CEQ 2000 Dye Terminator Cycle Sequence system --- p.71 / Chapter 4.13.1.1 --- Materials --- p.71 / Chapter 4.13.1.2 --- Methods --- p.71 / Chapter 4.13.2 --- ABI PRISM´ёØ dRhodamine Terminator Cycle Sequencing system --- p.72 / Chapter 4.13.2.1 --- Materials --- p.72 / Chapter 4.13.2.2 --- Methods --- p.72 / Chapter 4.13.3 --- Homology search against computer databases --- p.73 / Chapter 4.14 --- Northern analysis of differentially expressed cDNA fragments --- p.73 / Chapter 4.14.1 --- Formaldehyde gel electrophoresis of total RNA --- p.74 / Chapter 4.14.1.1 --- Materials --- p.74 / Chapter 4.14.1.2 --- Methods --- p.74 / Chapter 4.14.2 --- Preparation of cDNA probes for hybridization --- p.74 / Chapter 4.14.2.1 --- PCR DIG labeling --- p.75 / Chapter 4.14.2.1.1 --- Materials --- p.75 / Chapter 4.14.2.1.2 --- Methods --- p.75 / Chapter 4.14.2.2 --- Random Prime cDNA DIG labeling --- p.75 / Chapter 4.14.2.2.1 --- Materials --- p.75 / Chapter 4.14.2.2.2 --- Methods --- p.76 / Chapter 4.14.3 --- Purification of DNA from agarose gel --- p.77 / Chapter 4.14.3.1 --- Materials --- p.77 / Chapter 4.14.3.2 --- Methods --- p.78 / Chapter 4.14.4 --- Hybridization --- p.78 / Chapter 4.14.4.1 --- Materials --- p.78 / Chapter 4.14.4.2 --- Methods --- p.73 / Chapter 4.14.5 --- Synthesis of mouse GAPDH probe from normalization --- p.80 / Chapter 4.14.5.1 --- Materials --- p.80 / Chapter 4.14.5.2 --- Methods --- p.80 / Chapter Chapter 5 --- Results --- p.82 / Chapter 5.1 --- Liver morphology --- p.82 / Chapter 5.2 --- Liver weight --- p.82 / Chapter 5.3 --- Serum triglyceride and cholesterol levels --- p.88 / Chapter 5.4 --- Confirmation of genotypes --- p.91 / Chapter 5.5 --- DNase I treatment --- p.91 / Chapter 5.6 --- FDD RT-PCR and band excision --- p.98 / Chapter 5.7 --- Reamplification of excised cDNA fragments --- p.111 / Chapter 5.8 --- Subcloning of reamplified cDNA fragments --- p.121 / Chapter 5.9 --- DNA sequencing of subcloned cDNA fragments --- p.124 / Chapter 5.10 --- Confirmation of the differentially expressed cDNA fragments by Northern blot analysis --- p.132 / Chapter 5.11 --- Temporal expression pattern of differentially expressed genes --- p.157 / Chapter 5.12 --- Tissue distribution pattern of differentially expressed genes --- p.171 / Chapter Chapter 6 --- Discussions --- p.183 / Chapter 6.1 --- "Lack of hepatomegaly, hypotriglyceridemia and hepatic nodule formation in PPARα (-/-) mice" --- p.184 / Chapter 6.2 --- "Identification of PPARα-dependent and Wy-14,643 responsive genes" --- p.185 / Chapter 6.3 --- Functional roles of the isolated cDNA fragments --- p.186 / Chapter 6.3.1 --- Fragments B14 and H4 --- p.187 / Chapter 6.3.2 --- Fragment H1 --- p.189 / Chapter 6.3.3 --- Fragment H5 --- p.192 / Chapter 6.3.4 --- Fragment H8 --- p.194 / Chapter 6.4 --- Temporal expression patterns of the isolated cDNA fragments --- p.196 / Chapter 6.5 --- Tissue distribution patterns of the isolated cDNA fragments --- p.197 / Chapter Chapter 7 --- Conclusions --- p.200 / Chapter Chapter 8 --- Future studies --- p.204 / Chapter 8.1 --- Subcloning and characterization of the other differentially expressed genes --- p.204 / Chapter 8.2 --- Overexpression and inhibition expression of specific genes --- p.204 / Chapter 8.3 --- Generating transgenic mice with target disruption of specific gene --- p.205 / References --- p.206
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_323194 |
| Date | January 2000 |
| Contributors | Lee, Wing Sum., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xxv, 226 leaves : ill. (some col., some mounted) ; 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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