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Relationship between serum corticosteroid level and telomere length/telomerase activity in spleen cells of Balb/c mice.January 2007 (has links)
Chiu, Wang Kei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 95-110). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Contents --- p.v / Acknowledgements --- p.viii / Abbreviations --- p.ix / List of tables --- p.xii / List of figures --- p.xii / Chapter 1. --- Literature Review --- p.1 / Chapter 1.1 --- Cell cycle and chromosome replication --- p.1 / Chapter 1.2 --- Telomere-associated proteins --- p.4 / Chapter 1.3 --- Telomere repair protein --- p.6 / Chapter 1.4 --- Function of telomere --- p.9 / Chapter 1.5 --- Telomerase --- p.10 / Chapter 1.6 --- Factors affecting telomere length --- p.14 / Chapter 1.7 --- Stress and telomere length --- p.14 / Chapter 1.8 --- Definition of Stress --- p.16 / Chapter 1.9 --- Central nervous system components involved in stress response --- p.17 / Chapter 1.10 --- Glucocorticoid --- p.17 / Chapter 1.11 --- Physiological effects under the activation of stress system --- p.19 / Chapter 1.12 --- Effects of chronic hyperactivation of the stress system --- p.20 / Chapter 1.13 --- Hypothesis --- p.23 / Chapter 2. --- Materials and Methods --- p.24 / Chapter 2.1 --- Materials --- p.24 / Chapter 2.2 --- Experiment animals --- p.25 / Chapter 2.3 --- Methods --- p.26 / Chapter 2.3.1 --- Treatment schedule --- p.26 / Chapter 2.3.2 --- Organ extraction and serum preparation --- p.27 / Chapter 2.4 --- Serum corticosteroid assay --- p.27 / Chapter 2.5 --- Telomere length assay --- p.30 / Chapter 2.5.1 --- Genomic DNA extraction --- p.30 / Chapter 2.5.2 --- Genomic DNA digestion --- p.31 / Chapter 2.5.3 --- Southern blotting procedure --- p.32 / Chapter 2.6 --- Telomeric Repeat Amplification Protocol (TRAP) assay --- p.37 / Chapter 2.6.1 --- Extract preparation and protein concentration quanititation --- p.38 / Chapter 2.6.2 --- Real-time PCR reaction --- p.39 / Chapter 2.6.3 --- Melt Curve --- p.41 / Chapter 2.7 --- Detection of mouse telomerase reverse transcriptase component (mTERT) mRNA expression by reverse transcriptase- polymerase chain reaction (RT-PCR) --- p.42 / Chapter 2.7.1 --- Total RNA extraction --- p.42 / Chapter 2.7.2 --- RT-PCR --- p.44 / Chapter 2.7.3 --- Agarose gel electrophoresis of RT-PCR products --- p.45 / Chapter 2.8 --- Detection of mouse TERT (mTERT) by Western blotting --- p.46 / Chapter 2.8.1 --- Nuclear protein extraction --- p.46 / Chapter 2.8.2 --- Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.47 / Chapter 2.9 --- Statistics --- p.49 / Chapter 3. --- Results --- p.51 / Chapter 3.1 --- Body and spleen weights of study animals --- p.51 / Chapter 3.2 --- Serum corticosterone level --- p.54 / Chapter 3.3 --- Telomere lengths of spleen --- p.54 / Chapter 3.4 --- Telomerase activity of spleen tissue --- p.58 / Chapter 3.5 --- Correlation between serum corticosterone level and telomere length --- p.65 / Chapter 3.6 --- Correlation between serum corticosterone level and telomerase activity --- p.66 / Chapter 3.7 --- Correlation between telomere length and telomerase activity --- p.67 / Chapter 3.8 --- Detection of telomerase reverse transcriptase component (mTERT) mRNA expression by RT-PCR --- p.69 / Chapter 3.9 --- Detection of mTERT by Western blotting --- p.72 / Chapter 3.10 --- Correlation between serum corticosterone level and mTERT mRNA /protein expression --- p.75 / Chapter 4. --- Discussion --- p.77 / Chapter 4.1 --- Serum corticosterone level in mice --- p.78 / Chapter 4.2 --- Mice body weight and spleen weight --- p.80 / Chapter 4.3 --- Telomere lengths in the spleen tissue --- p.83 / Chapter 4.4 --- Telomerase activity in spleens --- p.84 / Chapter 4.5 --- Correlation between serum corticosterone level and telomere length --- p.87 / Chapter 4.6 --- Correlation between serum corticosterone level and telomerase activity --- p.89 / Chapter 4.7 --- Mouse telomerase reverse transcriptase component (mTERT) mRNA expression --- p.89 / Chapter 4.8 --- Expression of mTERT protein --- p.90 / Chapter 4.9 --- Conclusion --- p.92 / Chapter 5. --- References --- p.95 / Chapter 6. --- Appendix --- p.111
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