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Studies of the control of VEGF expression in testicular cell lines and in the testis.

Sy Chun Choi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 123-160). / Abstracts in English and Chinese. / ABSTRACT --- p.i / 摘要 --- p.iv / ACKNOWLEDGEMENT --- p.vi / Chapter 1. --- Introduction / Chapter 1.1 --- General review of the testis --- p.1 / Chapter 1.1.1 --- Structure and function of the testis --- p.1 / Chapter 1.1.2 --- Testicular vasculature --- p.2 / Chapter 1.1.3 --- Testicular angiogenesis --- p.3 / Chapter 1.2 --- Vascular endothelial growth factor (VEGF) --- p.4 / Chapter 1.2.1 --- Discovery of VEGF --- p.4 / Chapter 1.2.2 --- Organization of VEGF --- p.4 / Chapter 1.2.3 --- Properties of the VEGF isoforms --- p.5 / Chapter 1.2.4 --- VEGF receptors --- p.6 / Chapter 1.2.5 --- Functions of VEGF --- p.8 / Chapter 1.3 --- VEGF in the testis --- p.10 / Chapter 1.3.1 --- Localization of VEGF and VEGF receptors in the testis --- p.10 / Chapter 1.3.2 --- Postulated functions of VEGF in the testis --- p.11 / Chapter 1.4 --- Regulation of VEGF --- p.11 / Chapter 1.4.1 --- "VEGF, hypoxia and the testis" --- p.11 / Chapter 1.4.2 --- "VEGF, nitric oxide and the testis" --- p.14 / Chapter 1.4.3 --- Cadmium-induced testicular toxicity --- p.16 / Chapter 1.4.4 --- "VEGF, glucocorticoids and the testis" --- p.17 / Chapter 1.4.5 --- Hormonal regulation of VEGF expression 226}0ؤimportance of LH --- p.19 / Chapter 1.4.6 --- VEGF and Leydig cell - macrophage interaction --- p.21 / Chapter 1.5 --- Aims of the present study --- p.24 / Chapter 2. --- Materials and methods / Chapter 2.1 --- Animals --- p.26 / Chapter 2.1.1 --- Spermatic cord torsion --- p.26 / Chapter 2.1.2 --- Cadmium chloride treatment --- p.27 / Chapter 2.1.3 --- Leydig cell depletion and cadmium chloride treatment --- p.28 / Chapter 2.1.4 --- Dexamethasone pretreatment and cadmium chloride injection --- p.28 / Chapter 2.1.5 --- hCG injection --- p.29 / Chapter 2.2 --- Immunohistochemistry --- p.29 / Chapter 2.2.1 --- Perfusion fixation of the testes --- p.29 / Chapter 2.2.2 --- Processing of the testes for histological section --- p.29 / Chapter 2.2.3 --- Immunohistochemical staining for VEGF --- p.30 / Chapter 2.2.4 --- Photomicrograph --- p.32 / Chapter 2.3 --- Cell cultures --- p.32 / Chapter 2.3.1 --- Cell lines of mouse TM3 Leydig cells and TM4 Sertoli cells --- p.32 / Chapter 2.3.2 --- Tumour cell line of mouse MLTC-1 Leydig cells --- p.33 / Chapter 2.4 --- Cell treatments --- p.33 / Chapter 2.4.1 --- Hypoxic treatment --- p.34 / Chapter 2.4.2 --- Cadmium chloride treatment --- p.36 / Chapter 2.4.3 --- hCG treatment --- p.37 / Chapter 2.4.4 --- Activators of second messenger systems --- p.37 / Chapter 2.4.5 --- Effect of pro-inflammatory cytokines and angiogenic growth factors --- p.38 / Chapter 2.5 --- Preparation of cDNA probes --- p.39 / Chapter 2.5.1 --- VEGF cDNA probe preparation --- p.39 / Chapter 2.5.2 --- P-actin cDNA probe preparation --- p.42 / Chapter 2.5.3 --- Purification of PCR products --- p.44 / Chapter 2.5.4 --- Confirmation of PCR products --- p.45 / Chapter 2.5.5 --- cDNA probe labeling --- p.46 / Chapter 2.6 --- RNA extraction --- p.46 / Chapter 2.6.1 --- Extraction of total RNA from testicular cell lines --- p.46 / Chapter 2.6.2 --- Extraction total RNA from testicular tissues --- p.50 / Chapter 2.7 --- Northern blot analysis --- p.51 / Chapter 2.7.1 --- Measurement of total RNA concentration --- p.51 / Chapter 2.7.2 --- RNA gel electrophoresis --- p.52 / Chapter 2.7.3 --- Transfer of RNA to membrane --- p.53 / Chapter 2.7.4 --- Hybridization with [α-32P]dCTP-labelled probes --- p.53 / Chapter 2.7.5 --- Autoradiography and densitometric quantification --- p.54 / Chapter 2.8 --- Data and statistical analysis --- p.55 / Chapter 3. --- Results / Chapter 3.1 --- Effects of hypoxia and cobalt chloride treatment on VEGF expression in TM3 and TM4 cells --- p.57 / Chapter 3.2 --- Effects of testicular torsion on VEGF expression in adult rat testes --- p.61 / Chapter 3.3 --- Antagonism of hypoxic induction of VEGF expression in TM3 cells by nitric oxide --- p.66 / Chapter 3.4 --- Effect of cadmium on VEGF expression in TM3 and TM4 cells --- p.66 / Chapter 3.5 --- Effect of dexamethasone on Cd-induced increase in VEGF expression in TM3 cells --- p.73 / Chapter 3.6 --- Effect of cadmium treatment on VEGF expression in the adult rat testes --- p.73 / Chapter 3.7 --- Effect of Leydig cell depletion on basal and Cd-induced expression of VEGF in adult rat testes --- p.76 / Chapter 3.8 --- Effect of dexamethasone on basal and Cd-induced expression of VEGF in adult rat testes --- p.76 / Chapter 3.9 --- "Effects of hCG,forskolin and phorbol ester on VEGF expression in TM3 and TM4 cells" --- p.79 / Chapter 3.10 --- Effect of hCG on VEGF expression in MLTC-1 cells --- p.92 / Chapter 3.11 --- "Effect of EL-lα, IL-1β, IL-6, TNF- α and TNF- β on VEGF expression in TM3 cells" --- p.92 / Chapter 3.12 --- Effect of bFGF and TGF- β1 on VEGF expression in TM3 cells --- p.102 / Chapter 4. --- Discussion / Chapter 4.1 --- Upregulation of VEGF expression in TM3 and TM4 cells by hypoxia and cobalt chloride --- p.108 / Chapter 4.2 --- Effect of testicular torsion on VEGF expression in adult rat testes --- p.110 / Chapter 4.3 --- Antagonism of hypoxic induction of VEGF expression in TM3 cells by nitric oxide --- p.111 / Chapter 4.4 --- "Effect of cadmium on VEGF mRNA levels in TM3 and TM4 cells, and in adult rat testes" --- p.113 / Chapter 4.5 --- "Effect of hCG,forskolin and phorbol ester on VEGF expression in TM3 and TM4 cells" --- p.116 / Chapter 4.6 --- Effect of cytokines and growth factors on VEGF expression in TM3 cells --- p.119 / Chapter 5. --- References --- p.123

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_322874
Date January 1999
ContributorsSy, Chun Choi., Chinese University of Hong Kong Graduate School. Division of Physiology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, vi, 160 leaves : ill. (some col., some mounted) ; 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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