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Regulation of chloride secretion by P2Y receptors in polarized human bronchial epithelia, 16HBE14o-.January 2007 (has links)
Wong, Miu Fong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 140-152). / Abstracts in English and Chinese. / DECLARATION --- p.i / ACKNOWLEDGEMENT --- p.ii / ABBREVIATIONS --- p.iii / ABSTRACT IN ENGLISH --- p.iv / ABSTRACT IN CHINESE --- p.vii / TABLE OF CONTENTS --- p.ix / LIST OF FIGURES --- p.xii / LIST OF TABLES --- p.xviii / Chapter CHAPTER I - --- INTRODUCTION / Chapter 1.1 --- Regulation of human airway surface liquid --- p.1 / Chapter 1.2 --- Sodium reabsorption and chloride secretion in airway epithelium --- p.3 / Chapter 1.3 --- Purinergic receptors --- p.7 / Chapter 1.4 --- P2Y receptors in epithelial cells --- p.11 / Chapter 1.5 --- Autocrine or paracrine regulation of ion transport in epithelial cells --- p.13 / Chapter 1.6 --- Signaling pathways underlying the regulation of ion transport by P2Y receptors stimulation --- p.16 / Chapter 1.7 --- The therapeutic potential of P2Y receptors in treating cystic fibrosis --- p.18 / Chapter 1.8 --- Particular interest on P2Y6 receptor as potential target for treatment of cystic fibrosis --- p.21 / Chapter 1.9 --- Properties of 16HBE14o- cell line --- p.23 / Chapter 1.10 --- Objectives of the present experiments --- p.25 / Chapter CHAPTER II - --- MATERIALS AND METHODS / Chapter 2.1 --- Solutions and Chemicals --- p.26 / Chapter 2.2 --- Cell culture --- p.28 / Chapter 2.3 --- Simultaneous measurement of short-circuit current (Isc) and intracellular calcium concentration ([Ca2+ ])i --- p.29 / Chapter 2.3.1 --- Preparation of 16HBE14o- cells for simultaneous measurement of Isc and [Ca2+]i --- p.29 / Chapter 2.3.2 --- Measurement of Isc and transepithelial resistance with Ussing chamber --- p.32 / Chapter 2.3.3 --- Simultaneous measurement of Isc and [Ca2+]i --- p.35 / Chapter 2.4 --- Measurement of protein kinase A activity --- p.38 / Chapter 2.5 --- Data analysis --- p.39 / Chapter CHAPTER III - --- RESULTS / Chapter 3.1 --- Apical and basolateral application of P2Y agonists induced Isc and [Ca2+]i responses in 16HBE14o- cells --- p.40 / Chapter 3.1.1 --- Effect of apical and basolateral application of ATP on Isc and [Ca2+̐]ư --- p.40 / Chapter 3.1.2 --- Effect of apical and basolateral application of UTP on Isc and [Ca2+̐]ư --- p.45 / Chapter 3.1.3 --- Effect of apical and basolateral application of UDP on Isc and [Ca2+̐]ư --- p.50 / Chapter 3.1.4 --- "Summary of the effects of apical and basolateral application of ATP, UTP and UDP on Isc and [Ca2+̐]ư" --- p.55 / Chapter 3.2 --- Ionic mechanisms underlying the effect of apical and basolateral UDP on 16HBE14o- cells --- p.56 / Chapter 3.2.1 --- Differential effect of apical and basolateral UDP on Isc --- p.56 / Chapter 3.2.2 --- Effect of various apical CI- channel blockers on Isc response induced by apical and basolateral application of UDP --- p.59 / Chapter 3.2.3 --- Effect of various basolateral K+ channel blockers on Isc response induced by apical and basolateral application of UDP --- p.83 / Chapter 3.3 --- Involvement of other signaling molecules or pathways in regulation of the chloride secreting response evoked by apical and basolateral UDP --- p.108 / Chapter 3.3.1 --- Effect of apical and basolateral UDP on PKA activity --- p.109 / Chapter 3.3.2 --- Effect of PKC inhibitors on Isc response induced by apical and basolateral application of UDP --- p.111 / Chapter CHAPTER IV - --- DISCUSSION / Chapter 4.1 --- Simultaneous measurement of Isc and [Ca2+ ̐]ư upon apical and basolateral application of P2Y agonists in 16HBE14o- cells --- p.125 / Chapter 4.2 --- Ionic mechanism underlying the effect of apical and basolateral UDP on 16HBE14o- cells --- p.128 / Chapter 4.2.1 --- Possible ionic mechanism for chloride secretion mediated by apical P2Y6 receptors --- p.131 / Chapter 4.2.2 --- Possible ionic mechanism for chloride secretion mediated by basolateral P2Y6 receptors --- p.133 / Chapter 4.3 --- Involvement of other possible signaling molecules or pathway underlying the action of apical and basolateral UDP --- p.135 / Chapter 4.4 --- Summary --- p.138 / Chapter CHAPTER V - --- REFERENCES --- p.140 / Publications --- p.153
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