No description available.

Chan Ling Nga. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 143-153). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.vi / Table of Contents --- p.vii / List of Figures --- p.xi / List of Tables --- p.xiv / Abbreviations --- p.xv / Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- The Human Endometrium --- p.1 / Chapter 1.1.1 --- The Structure of the Endometrium --- p.1 / Chapter 1.1.2 --- Cyclic Changes in the Endometrium --- p.1 / Chapter 1.1.3 --- Physiological Roles of the Endometrium --- p.5 / Chapter 1.1.4 --- Roles of Luminal Epithelium in Implantation --- p.5 / Chapter 1.1.5 --- Exocrine Functions of the Endometrial Epithelium --- p.6 / Chapter 1.2 --- Review of Epithelial Ion Channels --- p.8 / Chapter 1.2.1 --- Epithelial Na+ Channels (ENaC) in Absorbing Epithelia --- p.9 / Chapter 1.2.2 --- Epithelial C1- Channels in Secretory Epithelia --- p.13 / Chapter 1.2.3 --- Na+ and C1- Channels in Endometrial Epithelia --- p.15 / Chapter 1.3 --- Review of the Intracellular Signal Transduction Pathways --- p.15 / Chapter 1.3.1 --- The cAMP-Mediated Signal Transduction Pathway --- p.17 / Chapter 1.3.2 --- The cAMP-Mediated Chloride Channels in Epithelial Cells --- p.17 / Chapter 1.3.3 --- Ca2+-Dependent Signal Transduction Pathway --- p.21 / Chapter 1.4 --- Physiological Roles of some Neurohormonal Agents in Uterine Functions: Selected Examples --- p.23 / Chapter 1.4.1 --- Roles of Adrenaline on the Endometrial Ion Transport --- p.23 / Chapter 1.4.2 --- Prostaglandin (PG) E2 and PGF2α --- p.24 / Chapter 1.4.3 --- Biological Effect of Extracellular Nucleotides --- p.26 / Chapter 1.5 --- Objective of this Study --- p.28 / Chapter 2 --- Materials and Methods --- p.31 / Chapter 2.1 --- Materials --- p.31 / Chapter 2.1.1 --- Culture Media and Enzymes --- p.31 / Chapter 2.1.2 --- Drugs --- p.31 / Chapter 2.1.3 --- Chemicals --- p.32 / Chapter 2.1.4 --- Experimental Tissues and Animals --- p.32 / Chapter 2.2 --- Preparations --- p.32 / Chapter 2.2.1 --- Previous Support for Cell Growth --- p.32 / Chapter 2.2.2 --- Growth Medium --- p.33 / Chapter 2.2.3 --- Culture of Mouse Endometrial Epithelial Cells --- p.35 / Chapter 2.2.4 --- Solutions for the Short-Circuit Current Measurements --- p.36 / Chapter 2.2.5 --- Solutions for the Patch-Clamp Experiments --- p.38 / Chapter 2.2.6 --- Running Buffers for RNA and DNA Gel Electrophoresis --- p.39 / Chapter 2.2.7 --- UTP-free UDP --- p.40 / Chapter 2.2.8 --- Electrodes for the Short-Circuit Current Measurement --- p.40 / Chapter 2.3 --- Protocols --- p.41 / Chapter 2.3.1 --- Characterization of Neurohormonal Agents-induced Ion Channels --- p.41 / Chapter 2.3.2 --- Possible Interaction between CFTR and ENaC --- p.41 / Chapter 2.3.3 --- Characterization of Pyrimidinoceptors-mediated Conductances --- p.42 / Chapter 2.4 --- Methods of Measurements --- p.42 / Chapter 2.4.1 --- The Patch-Clamp Technique --- p.42 / Chapter 2.4.1.1 --- The Patch-Clamp Setup --- p.43 / Chapter 2.4.1.2 --- Shielding and Grounding --- p.45 / Chapter 2.4.1.3 --- Pipette Fabrication --- p.45 / Chapter 2.4.1.4 --- Pipette Holder and Electrodes --- p.48 / Chapter 2.4.1.5 --- Experimental Procedures --- p.49 / Chapter 2.4.1.6 --- Signal Recording and Data Acquisition --- p.54 / Chapter 2.4.1.7 --- Data Analysis --- p.54 / Chapter 2.4.2 --- The Short-Circuit Current Technique --- p.55 / Chapter 2.4.2.1 --- The Short-Circuit Current Setup --- p.56 / Chapter 2.4.2.2 --- Experimental Procedures --- p.56 / Chapter 2.4.2.3 --- Data Analysis --- p.61 / Chapter 2.4.3 --- Reverse Transciption - Polymerase Chain Reaction (RT-PCR) --- p.61 / Chapter 2.4.3.1 --- RNA Isolation --- p.61 / Chapter 2.4.3.2 --- RNA Gel Electrophoresis --- p.62 / Chapter 2.4.3.3 --- Reverse Transcription (RT) --- p.63 / Chapter 2.4.3.4 --- Polymerase Chain Reaction (PCR) --- p.64 / Chapter 2.4.3.5 --- DNA Gel Electrophoresis --- p.66 / Chapter 2.4.4 --- Statistical Analysis --- p.66 / Chapter 3 --- Results --- p.67 / Chapter 3.1 --- Activation of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) in Response to Hormonal Stimuli --- p.67 / Chapter 3.2 --- Inhibition of Na+ Absorption by CFTR --- p.89 / Chapter 3.3 --- Pyrimidinoceptors-activated Ca2+-dependent C1- Conductance --- p.111 / Chapter 4 --- General Discussions --- p.132 / Appendix --- p.140 / Chapter A --- RNA Isolation --- p.140 / Chapter B --- Reverse Transcription --- p.141 / Chapter C --- Polymerase Chain Reaction --- p.142 / References --- p.143

A mouse in vitro co-culture model between endometrial epithelial cells (EEC) and peripheral blood lymphocytes and monocytes (PBLM) immune cells was established, and Ct lipopolysaccharide (LPS) was added to the cells to mimic Ct bacterial infection and to stimulate an immune response. This model enabled us to study the cross-talk between EEC and PBLM and the physiological changes that occur in the endometrium upon Ct LPS stimulation. Results showed that EEC-PBLM co-culture and Ct LPS stimulation caused changes in transepithelial resistance (TER) as well as the expression and function of the cystic fibrosis transmembrane conductance regulator (CFTR) ion channel of epithelial cells. CFTR gene transcription was up-regulated at early hours after Ct LPS stimulation, while its channel activity was down-regulated at later hours. These results suggested the possible involvement of CFTR acting as a receptor for the internalization of Ct, which may ultimately lead to the disappearance of CFTR on the apical membrane. The EEC-PBLM co-culture showed that cross-talk was important for host defense in the endometrium. Direct cross-talk by cell-cell contact between EEC and PBLM was vital for immune cell survival as well as strengthening epithelials' barrier function. (Abstract shortened by UMI.) / Chlamydia trachomatis (Ct) infection is one of the most prevalent causes for sexually transmitted disease (STD) in the female reproductive tract. Ct is unique in that it is a bacterium, but infects and replicates like a virus inside a host cell. Ct infection can lead to a variety of reproductive diseases, such as pelvic inflammatory diseases (PID), tubule scarring, salpingitis, endometriosis, ectopic pregnancy and infertility. Effective immune defense in the uterus is necessary to eliminate these bacteria and to ensure optimal uterine environment for sperm motility, fertilization, and embryo implantation to occur. The immune system of the endometrium responds to Ct infection by the recruitment of many types of leukocytes, such as T-lymphocytes, B-lymphocytes, monocytes, macrophages and neutrophils, to the site of infection. Cross-talk between endometrial epithelial cells and immune cells may alter the activities of epithelial cells causing changes in channels function and anion secretion. / Ho Alice. / "August 2005." / Adviser: Chan Hsiao Chang. / Source: Dissertation Abstracts International, Volume: 67-07, Section: B, page: 3532. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 155-167). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract in English and Chinese. / School code: 1307.