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Studies of localization and expression of angiopoietin in the testis.

Wong Chun Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 149-160). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Abbreviations --- p.v / Acknowledgement --- p.x / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- General review of angiogenesis --- p.1 / Chapter 1.1.1 --- Angiogenesis in development and growth --- p.1 / Chapter 1.1.2 --- The process of angiogenesis --- p.2 / Chapter 1.1.3 --- Types of factors controlling angiogenesis --- p.3 / Chapter 1.2 --- Roles of VEGF and its receptors in the regulation of angiogenesis --- p.6 / Chapter 1.2.1 --- VEGF --- p.6 / Chapter 1.2.2 --- VEGF receptors --- p.8 / Chapter 1.2.3 --- Regulation of VEGF expression by hypoxia and nitric oxide… --- p.10 / Chapter 1.2.4 --- Signal transduction mechanisms of VEGFR-1 and VEGFR-2 --- p.12 / Chapter 1.2.5 --- Anti-apoptotic effect ofVEGF on endothelial cells as a result of signal transduction of VEGFR-2 --- p.14 / Chapter 1.3 --- Angiopoietins --- p.15 / Chapter 1.3.1 --- Angiopoietin 1 (Ang-1) --- p.16 / Chapter 1.3.2 --- Angiopoietin 2 (Ang-2) --- p.19 / Chapter 1.3.3 --- Angiopoietins 3 and 4 (Ang-3 and Ang-4) --- p.24 / Chapter 1.4 --- "Interaction among VEGF, angiopoietin and Tie in the maintenance of vasculature" --- p.25 / Chapter 1.5 --- Tyrosine kinase with immunoglobulin and EGF factor homology domains - Tie 1 and Tie 2 --- p.28 / Chapter 1.6 --- Angiopoietin expression in female reproductive tissues (ovary) --- p.33 / Chapter 1.7 --- Testicular angiogenesis --- p.37 / Chapter 1.8 --- Aims of the present study --- p.38 / Chapter Chapter 2 --- Materials and methods / Chapter 2.1 --- Preparation of primary cells from rat testes --- p.40 / Chapter 2.1.1 --- Sertoli cell preparation --- p.40 / Chapter 2.1.2 --- Germ cell preparation --- p.41 / Chapter 2.1.3 --- Interstitial cell and Leydig cell preparation --- p.43 / Chapter 2.2 --- Cell cultures --- p.45 / Chapter 2.2.1 --- Reagents and cell lines --- p.45 / Chapter 2.2.2 --- Cell lines of mouse TM3 Leydig cells and TM4 Sertoli cells --- p.45 / Chapter 2.2.3 --- Mouse MLTC-1 Leydig tumour cells --- p.46 / Chapter 2.2.4 --- Rat R2C Leydig tumour cells --- p.46 / Chapter 2.2.5 --- Rat LC540 Leydig tumour cells --- p.47 / Chapter 2.2.6 --- "Rat C6 glioma cells.............," --- p.47 / Chapter 2.3 --- "Analyses of Angiopoietin 1, Angiopoietin 2, Angiopoietin3, Tie 1 receptor, and Tie 2 receptor mRNA in testicular cell lines and testicular tissues" --- p.48 / Chapter 2.3.1 --- Extraction of total RNA from testicular cell lines and testicular tissues --- p.48 / Chapter 2.3.2 --- Quantitation of total RNA --- p.50 / Chapter 2.3.3 --- First strand cDNA synthesis by reverse transcription (RT) --- p.51 / Chapter 2.3.4 --- Normalization of the amounts of cDNA usedin polymerase chain reaction (PCR) --- p.52 / Chapter 2.3.5 --- Polymerase chain reaction (PCR) --- p.53 / Chapter 2.3.6 --- Purification of PCR products --- p.65 / Chapter 2.3.7 --- Confirmation of PCR product authenticity by automated DNA sequencing --- p.66 / Chapter 2.4 --- Western blot analysis --- p.68 / Chapter 2.4.1 --- Preparation of cell lysates from primary testicular cells and testicular cell lines --- p.68 / Chapter 2.4.2 --- Preparation of mouse testicular tissue and adult rat testicular tissue lysates --- p.68 / Chapter 2.4.3 --- Determination of protein concentration --- p.69 / Chapter 2.4.4 --- Reagents for Western blot analysis --- p.70 / Chapter 2.4.5 --- Preparation of protein samples and markers for Western blot analysis --- p.71 / Chapter 2.4.6 --- Sodium dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.72 / Chapter 2.4.7 --- Transfer of proteins to membrane --- p.74 / Chapter 2.4.8 --- Blocking of the membrane --- p.74 / Chapter 2.4.9 --- Immunoblotting --- p.75 / Chapter 2.5 --- "Immunohistochemical staining for Ang-1, Ang-2,Ang-3, Tie 1 and Tie 2 in rat testes" --- p.78 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Expression of Ang-1 and Ang-1 alternatively spliced transcripts in the testis and other testicular cell types --- p.81 / Chapter 3.1.1 --- Detection of Ang-1 expression in the testis and and testicular cell types by nested PCR --- p.81 / Chapter 3.1.2 --- Detection of Ang-1 expression in testicular cell lines by nested PCR --- p.82 / Chapter 3.1.3 --- Sequence analysis of Ang-1 transcript amplified from adult rat testis --- p.84 / Chapter 3.1.4 --- Detection of alternatively spliced species of Ang-1 mRNA in the testis and other testicular cell lines --- p.87 / Chapter 3.2 --- Expression of Ang-2 and Ang-2 isoforms in the testis and various testicular cell types --- p.94 / Chapter 3.2.1 --- Detection of Ang-2 expression in the testis and testicular cell types by nested PCR --- p.94 / Chapter 3.2.2 --- Detection of Ang-2 expression in testicular cell lines by nested PCR --- p.96 / Chapter 3.2.3 --- Sequence analysis of Ang-2 transcript amplified from adult rat testis --- p.98 / Chapter 3.2.4 --- Detection of the expression of Ang-2 isoforms in adult rat testis --- p.99 / Chapter 3.3 --- Expression of Ang-3 in the testis and testicular cell types --- p.103 / Chapter 3.3.1 --- Detection of Ang-3 expression in the testis and primary testicular cells by RT-PCR --- p.103 / Chapter 3.3.2 --- Detection of Ang-3 expression in testicular cell lines by RT-PCR --- p.105 / Chapter 3.3.3 --- Sequence analysis of Ang-3 transcripts amplified from TM4 mouse Sertoli cells and adult rat testis --- p.105 / Chapter 3.4 --- Expression of Tie 1 and Tie 2 in the testis and testicular blood vessel --- p.110 / Chapter 3.4.1 --- Detection of Tie 1 and Tie 2 expression in the testis and rat testicular blood vessel by RT-PCR --- p.110 / Chapter 3.4.2 --- Sequence analysis of Tie 1 transcripts amplified from adult rat testis and rat testicular blood vessel --- p.113 / Chapter 3.4.3 --- Sequence analysis of Tie 2 transcript amplified from rat testicular blood vessel --- p.113 / Chapter 3.5 --- "Western blot analysis of Ang-1 and Ang-2 expression in testicular tissues, primary testicular cells and cell lines" --- p.116 / Chapter 3.6 --- "Localization of Ang-1,Ang-2, Ang-4, Tie 1 and Tie2 proteins in adult rat testis by immunohistochemistry" --- p.122 / Chapter 3.7 --- "Comparison of angiopoietin expression patterns in testis using RT-PCR, Western immunoblotting and immunohistochemistry" --- p.128 / Chapter 3.8 --- Comparison of Tie 1 and Tie 2 expression patterns in testis using RT-PCR and immunohistochemistry --- p.128 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- "Expression of Ang-1 mRNA and protein in adult rat testis, mouse testis, rat testicular blood vessel, primary testicular cells and testicular cell lines" --- p.131 / Chapter 4.2 --- "Expression of Ang-2 mRNA and protein in adult rat testis, mouse testis, rat testicular blood vessel, primary testicular cells and testicular cell lines" --- p.136 / Chapter 4.3 --- "Expression of Ang-3 mRNA and protein in adult rat testis, mouse testis, rat testicular blood vessel, primary testicular cells and testicular cell lines" --- p.141 / Chapter 4.4 --- Expression of Tie 1 and Tie 2 mRNAs and proteinsin adult rat testis and rat testicular blood vessel --- p.143 / Chapter 4.5 --- Conclusion --- p.145 / Chapter 4.6 --- Future work --- p.146 / Chapter Chapter 5 --- References --- p.149

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324114
Date January 2001
ContributorsWong, Chun Yan., Chinese University of Hong Kong Graduate School. Division of Physiology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, x, 160 leaves : ill. (some col.) ; 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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