by Wan Sze Ping. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 172-185). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.v / Publications --- p.vi / Abbreviations --- p.vii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1. --- Historical Background --- p.2 / Chapter 1.2. --- Origin and heterogeneity of mast cells --- p.2 / Chapter 1.3. --- Mast cell mediators --- p.4 / Chapter 1.3.1. --- Preformed mediators --- p.5 / Chapter 1.3.2. --- Newly synthesised mediators --- p.6 / Chapter 1.4. --- Mast cell activation --- p.8 / Chapter 1.4.1. --- Antigenic/Immunological pathway --- p.8 / Chapter 1.4.1.1. --- Aggregation of IgE Receptors (FcsRI) --- p.9 / Chapter 1.4.1.2. --- Biochemical events following FcsRI aggregation --- p.10 / Chapter 1.4.1.3. --- Phospholipase C activation --- p.11 / Chapter 1.4.1.4. --- Phospholipase A2 activation --- p.12 / Chapter 1.4.1.5. --- Adenylate cyclase activation --- p.14 / Chapter 1.4.1.6. --- Role of calcium --- p.15 / Chapter 1.5. --- Roles of chloride channels in mast cell activation --- p.18 / Chapter 1.6. --- Aims of the study --- p.20 / Chapter Chapter 2 --- Materials and Methods --- p.23 / Chapter 2.1. --- Materials --- p.24 / Chapter 2.1.1. --- Secretagogues --- p.24 / Chapter 2.1.2. --- Anti-allergic compounds --- p.24 / Chapter 2.1.3. --- Chloride channel blockers --- p.24 / Chapter 2.1.4. --- Materials for buffers --- p.25 / Chapter 2.1.5. --- Materials for rat sensitisation --- p.25 / Chapter 2.1.6. --- Materials for histamine assay --- p.26 / Chapter 2.1.7. --- Materials for calcium measurement --- p.26 / Chapter 2.1.8. --- Miscellaneous --- p.27 / Chapter 2.2. --- Buffers --- p.28 / Chapter 2.3. --- Stock solutions of drugs --- p.30 / Chapter 2.4. --- Source of mast cells --- p.31 / Chapter 2.4.1. --- Animals --- p.31 / Chapter 2.4.2. --- Sensitization of animals --- p.31 / Chapter 2.4.3. --- Isolation of rat peritoneal mast cells --- p.32 / Chapter 2.4.4. --- Purification of rat peritoneal mast cells --- p.32 / Chapter 2.4.5. --- Cell counting --- p.33 / Chapter 2.5. --- General protocol for histamine release --- p.35 / Chapter 2.6. --- Histamine assay --- p.35 / Chapter 2.7. --- Protocol for fura-2 fluorescence measurement of intracellular calcium --- p.39 / Chapter 2.7.1. --- Fura-2 cell loading --- p.39 / Chapter 2.7.2. --- Fura-2 fluorescence measurement by fluorescence microscope --- p.39 / Chapter 2.7.3. --- Calculation of intracellular calcium --- p.41 / Chapter 2.8. --- Protocol for 45Ca2+ influx measurement --- p.44 / Chapter 2.9. --- Measurement of 43Ca2+ influx --- p.45 / Chapter 2.10. --- Statistical analysis --- p.45 / Chapter Chapter 3 --- Effects of chloride depletion on mast cell activation --- p.46 / Chapter 3.1. --- Introduction --- p.47 / Chapter 3.1.1. --- Mechanism of non-immunological stimuli --- p.48 / Chapter 3.2. --- Materials and methods --- p.50 / Chapter 3.3. --- Results --- p.51 / Chapter 3.3.1. --- Effects of anti-IgE induced histamine release in chloride free buffers --- p.51 / Chapter 3.3.2. --- Effects of compound 48/80 induced histamine releasein chloride free buffers --- p.52 / Chapter 3.3.3. --- Effects of calcium ionophore A23187 induced histamine release in chloride free buffers --- p.53 / Chapter 3.3.4. --- Effects of ionomycin induced histamine release in chloride free buffers --- p.53 / Chapter 3.3.5. --- Effects of thapsigargin induced histamine releasein chloride free buffers --- p.53 / Chapter 3.3.6. --- Effects of Triton-X 100 induced histamine releasein chloride free buffers --- p.54 / Chapter 3.3.7. --- Effects of anti-IgE on the free intracellular calcium concentration of rat peritoneal mast cells in chloride free buffers --- p.54 / Chapter 3.3.8. --- Effects of compound 48/80 on the free intracellular calcium concentration of rat peritoneal mast cellsin chloride free buffers --- p.56 / Chapter 3.3.9. --- Effects of ionomycin on the free intracellular calcium concentration of rat peritoneal mast cellsin chloride free buffers --- p.58 / Chapter 3.3.10. --- Effects of thapsigargin on the free intracellular calcium concentration of rat peritoneal mast cellsin chloride free buffers --- p.59 / Chapter 3.4. --- Discussion --- p.86 / Chapter 3.5. --- Conclusion --- p.96 / Chapter Chapter 4 --- "DSCG and Chloride channel blockers (NPPB, DIDS, SITS, DPC, IAA-94 & Flufenamic acid)" --- p.97 / Chapter 4.1. --- Introduction --- p.98 / Chapter 4.1.1. --- Role of chloride channels --- p.98 / Chapter 4.1.1.1. --- Chloride channel classification --- p.99 / Chapter 4.1.1.2. --- Chloride channel blockers --- p.101 / Chapter 4.1.2. --- Aims of study --- p.106 / Chapter 4.2. --- Materials and methods --- p.107 / Chapter 4.3. --- Results --- p.108 / Chapter 4.3.1. --- Effects of DSCG and the test blockers on immunologically induced histamine release from rat peritoneal mast cells --- p.108 / Chapter 4.3.2. --- Effects of DSCG and the test blockers on compound48/80 induced histamine release from rat peritoneal mast cells --- p.110 / Chapter 4.3.3. --- Effects of DSCG and the test blockers on ionophore A23187 induced histamine release from rat peritoneal mast cells --- p.112 / Chapter 4.3.4. --- Effects of DSCG and the test blockers on thapsigargin induced histamine release from rat peritoneal mast cells --- p.113 / Chapter 4.3.5. --- Effects of the test blockers on Triton-X 100 induced histamine release from rat peritoneal mast cells --- p.114 / Chapter 4.3.6. --- Effects of DSCG and the test blockers on immunologically and non-immunologically induced 45Ca2+ uptake --- p.115 / Chapter 4.4. --- Discussion --- p.160 / Chapter 4.5. --- Conclusion --- p.167 / References --- p.172