by Chung Hoi Sing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 92-106). / Abstract also in Chinese. / ABSTRACT --- p.i / 摘要 --- p.iii / ACKNOWLEDGMENT --- p.v / Chapter 1. --- Introduction / Chapter 1.1 --- Angiogenesis in general --- p.1 / Chapter 1.1.1 --- The concept of angiogenesis --- p.1 / Chapter 1.1.2 --- The process of angiogenesis --- p.1 / Chapter 1.2 --- Measurement of angiogenesis --- p.3 / Chapter 1.2.1 --- In vivo assays --- p.3 / Chapter 1.2.2 --- In vitro assays --- p.5 / Chapter 1.3 --- Angiogenic factors --- p.6 / Chapter 1.4 --- Angiogenesis in the female reproductive system --- p.7 / Chapter 1.5 --- Evidence of hormonally-regulated angiogenesis in endocrine tissues --- p.10 / Chapter 1.5.1 --- Ovary --- p.10 / Chapter 1.5.2 --- Thyroid --- p.11 / Chapter 1.6 --- Angiogenesis in the testis --- p.12 / Chapter 1.6.1 --- Structure of testicular vasculature --- p.12 / Chapter 1.6.2 --- Angiogenic factors in the testis --- p.13 / Chapter 1.6.3 --- Vascular effects of hCG/LH in the testis --- p.17 / Chapter 1.6.4 --- Postnatal development of testicular vasculature --- p.17 / Chapter 1.7 --- Aims of the present study --- p.19 / Chapter 2. --- Materials and methods / Chapter 2.1 --- Animals --- p.20 / Chapter 2.2 --- Experimental design --- p.20 / Chapter 2.2.1 --- Testicular angiogenesis in adult rats - hormonal stimulation by hCG --- p.20 / Chapter 2.2.1.1 --- Changes with time after hCG treatment --- p.20 / Chapter 2.2.1.2 --- Effect of Leydig cell depletion --- p.22 / Chapter 2.2.1.3 --- Effect of Leydig cell suppression by subcutaneous testosterone-filled silastic implants --- p.22 / Chapter 2.2.1.4 --- Effect of testicular macrophage activation --- p.24 / Chapter 2.2.1.5 --- Effect of testicular macrophage depletion --- p.26 / Chapter 2.2.2 --- Developmental changes in testicular angiogenesis --- p.29 / Chapter 2.3 --- Perfusion of testes with fixative or Indian Ink --- p.29 / Chapter 2.4 --- Processing of the testes for histological sections --- p.30 / Chapter 2.5 --- Immunohistochemical staining for proliferating cell nuclear antigen (PCNA) --- p.31 / Chapter 2.6 --- Immunohistochemical staining for vascular endothelial growth factor --- p.32 / Chapter 2.7 --- Quantification of PCNA-positive endothelial cells --- p.33 / Chapter 2.8 --- Quantification of blood vessel density --- p.34 / Chapter 2.9 --- Estimation of intertubular area in testis section --- p.35 / Chapter 2.10 --- Preparation of liposome-entrapped dichloromethylene diphosphonate (Cl2MDP-lp) --- p.38 / Chapter 2.11 --- Radioimmunoassay of serum tsetosterone --- p.38 / Chapter 2.12 --- Statistical analyses --- p.40 / Chapter 3. --- Results / Chapter 3.1 --- hCG-induced increase in endothelial cell proliferation in adult rat testes --- p.41 / Chapter 3.1.1 --- Testicular histology --- p.41 / Chapter 3.1.2 --- Changes in the number of PCNA-positive endothelial cells --- p.41 / Chapter 3.1.3 --- Changes in blood vessel density --- p.44 / Chapter 3.1.4 --- Changes in testis weight and serum testosterone concentration --- p.44 / Chapter 3.2 --- Effect of Leydig cell depletion by ethane dimethane sulphonate (EDS) on hCG-induced endothelial cell proliferation in adult rat testes --- p.48 / Chapter 3.2.1 --- Testicular histology --- p.48 / Chapter 3.2.2 --- Changes in the number of PCNA-positive endothelial cells --- p.48 / Chapter 3.2.3 --- Changes in serum testosterone concentration and testis weight --- p.52 / Chapter 3.3 --- Effect ofLeydig cell suppression by testosterone-filled subcutaneous silastic implants on hCG-induced endothelial cell proliferation in adult rat testes --- p.54 / Chapter 3.3.1 --- "Changes in serum testosterone concentration, testis weight, and testicular intertubular area" --- p.54 / Chapter 3.3.2 --- Changes in the number of PCNA-positive endothelial cells --- p.58 / Chapter 3.3.3 --- Changes in the level of vascular endothelial growth factor (VEGF) immunoreactivity in the testis --- p.60 / Chapter 3.4 --- Effect of testicular macrophage activation by polystyrene latex beads on hCG-induced endothelial cell proliferation in adult rat testes --- p.60 / Chapter 3.4.1 --- Testicular histology --- p.60 / Chapter 3.4.2 --- Changes in the number of PCNA-positive endothelial cells --- p.63 / Chapter 3.4.3 --- Changes in testis weight and serum testosterone concentration --- p.65 / Chapter 3.5 --- Effect of testicular macrophage depletion by liposome-entrapped C12MDP treatment on hCG-induced endothelial cell proliferation in adult rat testes --- p.67 / Chapter 3.5.1 --- Testicular histology --- p.68 / Chapter 3.5.2 --- Changes in the number of PCNA-positive endothelial cells --- p.68 / Chapter 3.5.3 --- Changes in testis weight and serum testosterone --- p.72 / Chapter 3.6 --- Endothelial cell proliferation in rat testes during postnatal development --- p.74 / Chapter 3.6.1 --- Changes in the number of PCNA-positive endothelial cells --- p.74 / Chapter 3.6.2 --- Changes in blood vessel density --- p.74 / Chapter 3.6.3 --- Changes in testis weight and intertubular area of the testes --- p.77 / Chapter 4. --- Discussion / Chapter 4.1 --- hCG-induced endothelial cell proliferation and changes in blood vessel density --- p.79 / Chapter 4.2 --- Role of Leydig cells in hCG-induced endothelial cell proliferation in adult rat testes --- p.82 / Chapter 4.3 --- Role of testicular macrophages in hCG-induced endothelial cell proliferation in adult rat testes --- p.86 / Chapter 4.4 --- Testicular angiogenesis during postnatal development --- p.88 / Chapter 5. --- References --- p.92

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

Tao Kin Pong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 144-152). / Abstracts in English and Chinese. / Chapter i. --- Cover page --- p.1 / Chapter ii. --- Table of contents --- p.2 / Chapter iii. --- Abstract of thesis (English version and Chinese Version) --- p.4 / Chapter iv. --- Acknowledgements --- p.8 / Chapter v. --- Abbreviations --- p.9 / Chapter 1. --- Introduction / Interstitial tissue and Leydig cells --- p.11 / Seminiferous tubules --- p.11 / "Male germ cell line and spermatogenesis (Mitotic, Meiotic and Post-meiotic)" --- p.12 / Sertoli cells --- p.14 / Specialized organizations of junction present in testis --- p.15 / Dynamic nature of Sertoli-Sertoli & Sertoli-germ cell junctions --- p.16 / Role of proteases and protease inhibitors in male gametogenesis --- p.17 / Proteases and Proteases Inhibitors expressed in testis --- p.18 / Hormonal control of spermatogenesis --- p.19 / Hypothyroidism and testis development --- p.21 / Genes to be studied: / Proteases --- p.22 / Proteases Inhibitors --- p.27 / Other spermatogenesis related genes --- p.30 / Chapter 2. --- Objectives --- p.32 / Chapter 3. --- Materials and Methods / Animal treatments and tissue collection --- p.33 / RNA preparation --- p.34 / RT-PCRs --- p.35 / Real-time PCRs --- p.35 / Data manipulations and Statistics --- p.36 / Primer sequences used in this experiment --- p.37 / Chapter 4. --- Results / "Effect of neonatal hypothyroidism on developmental profile of body weight, absolute and relative testicular weight" --- p.40 / Developmental transcription profiles of genes under normal and hypothyroidism --- p.43 / Screening Data --- p.78 / Expression of non-spermatogenic genes at neonatal age --- p.88 / Responsiveness of gene transcription after thyroxin replacement --- p.89 / Changes of gene expression under different hypothyroid regimens --- p.97 / Chapter 5. --- Discussion / Changes in testicular size under hypothyroidism --- p.107 / Five patterns of transcription profile --- p.107 / "Suggestion on the role of ""MEIOTIC"" proteases and inhibitors" --- p.111 / "Suggestion on the role of ""POST-MEIOTIC"" proteases and inhibitors" --- p.113 / "Explanations on ""SOMATIC"" genes" --- p.114 / "Explanations on ""MITOTIC"" genes" --- p.115 / Explanations on the un-clustered pattern --- p.116 / Explanations on the age down-regulated group --- p.116 / Proposed clustering of genes according to their transcription profile --- p.117 / "Expression of some ""non-spermatogenic"" genes before puberty" --- p.123 / Neonatal hypothyroidism as a model for studying reproductive physiology --- p.125 / Different components of spermatogenesis --- p.127 / Chapter I. --- Roles of nuclear and chromatin related genes in assisting meiosis --- p.128 / Chapter II. --- Roles of specific transcription regulators in assisting gene selection --- p.129 / Chapter III. --- Role of signal transduction molecules for translation and activation --- p.131 / Chapter IV. --- Role of proteases and inhibitors for matrix and junctions dynamics --- p.132 / The somatic proteases and inhibitors system in the testis --- p.133 / Spermatogenic proteases and inhibitors system --- p.134 / Chapter V. --- Role of matrix and junctional molecules in intercellular interactions --- p.137 / Chapter VI. --- Role of cytoplasmic motors in cellular movement --- p.139 / Chapter 6. --- Conclusion / Proposed story of spermatogenesis - involvement of proteases and inhibitors --- p.140 / Future Direction --- p.141 / Chapter 7. --- References --- p.144