A biochemical study of cell death, apoptosis, in macrophages.

• Other Authors: Chan, Yee Man Elaine.
• Format: Others
• Language: English
• Published: Chinese University of Hong Kong 1995
• Subjects: Apoptosis; Macrophages > Chemistry; Nitric oxide; Cell death
• Online Access: http://library.cuhk.edu.hk/record=b5888421; http://repository.lib.cuhk.edu.hk/en/item/cuhk-320668

by Chan Yee Man Elaine. === Thesis (M.Phil.)--Chinese University of Hong Kong, 1995. === Includes bibliographical references (leaves 130-143). === Abstract --- p.I === Acknowledgements --- p.III === Abbreviations --- p.IV === Objectives of the study --- p.VII === Table of Contents --- p.VIII === Chapter Section 1 --- Introduction --- p.1 === Chapter 1.1 --- Necrosis vs Apoptosis --- p.2 === Chapter 1.2 --- Cell Death by Apoptosis --- p.4 === Chapter 1.3 --- The Biochemistry of Nitric Oxide --- p.9 === Chapter 1.4 --- Mechanisms of NO Action --- p.14 === Chapter 1.5 --- Signal Transduction Pathways to Apoptosis --- p.17 === Chapter 1.5.1 --- Regulation by Ca2+ --- p.17 === Chapter 1.5.2 --- Protein Kinase C --- p.20 === Chapter 1.5.3 --- cAMP --- p.21 === Chapter 1.5.4 --- Protein tyrosine kinase --- p.21 === Chapter 1.5.5 --- Ceramide --- p.22 === Chapter 1.5.6 --- pH --- p.23 === Chapter 1.5.7 --- Oxygen Radicals --- p.23 === Chapter 1.5.8 --- Anchorage Dependence and Extracellular Matrix --- p.24 === Chapter Section 2 --- Materials and Methods --- p.27 === Chapter 2.1 --- Materials --- p.28 === Chapter 2.1.1 --- Animal --- p.28 === Chapter 2.1.2 --- Cell line --- p.28 === Chapter 2.1.3 --- "Culture media, buffers and chemicals" --- p.28 === Chapter 2.1.4 --- Dye solutions --- p.30 === Chapter 2.1.5 --- Reagents and buffers for polyacrylamide gel electrophoresis (PAGE) --- p.31 === Chapter 2.1.6 --- Reagents and buffers for Western blotting --- p.32 === Chapter 2.1.7 --- Reagents and buffers for agarose gel electrophoresis --- p.33 === Chapter 2.2 --- Methods --- p.35 === Chapter 2.2.1 --- Cell culture --- p.35 === Chapter 2.2.2 --- [3H]-Thymidine incorporation --- p.35 === Chapter 2.2.3 --- MTT assay --- p.36 === Chapter 2.2.4 --- Determination of NO by Griess assay --- p.36 === Chapter 2.2.5 --- Observation of apoptotic morphology of cells by confocal laser scanning microscopy (CLSM) --- p.37 === Chapter 2.2.6 --- Determination of cell death induced by NO producing drugs --- p.38 === Chapter 2.2.7 --- Determination of cell death induced by Concanavalin A --- p.38 === Chapter 2.2.8 --- Determination of effect of nitric oxide synthase (NOS) inhibitor on cell death induced by Con A --- p.39 === Chapter 2.2.9 --- Determination of the requirement of Ca2+ in cell death induced by NO producing drugs --- p.39 === Chapter 2.2.10 --- Determination of the requirement of cGMP in cell death induced by NO producing drugs --- p.40 === Chapter 2.2.11 --- Determination of cell death induced by PKC activation and depletion --- p.40 === Chapter 2.2.12 --- Determination of effect of PKC depletion on cell death induced by NO producing drugs and Con A --- p.41 === Chapter 2.2.13 --- Observation of immunofluorescence by confocal laser scanning microscopy --- p.41 === Chapter 2.2.14 --- Preparation of protein samples for PAGE --- p.42 === Chapter 2.2.15 --- Polyacrylamide gel electrophoresis --- p.43 === Chapter 2.2.16 --- Western blotting of PKC --- p.44 === Chapter 2.2.17 --- Preparation of DNA samples from cells --- p.46 === Chapter 2.2.18 --- Agarose gel electrophoresis of DNA --- p.47 === Chapter 2.2.19 --- Statistical analysis --- p.48 === Chapter Section 3 --- Results --- p.49 === Chapter 3.1 --- Induction of apoptosis in macrophages by NO producing drugs --- p.50 === Chapter 3.2 --- Apoptosis induced by NO producing drugs was caused by NO --- p.54 === Chapter 3.3 --- Signal transduction pathways to the NO-induced cell death in macrophages --- p.66 === Chapter 3.3.1 --- Calcium ion --- p.66 === Chapter 3.3.2 --- cGMP --- p.68 === Chapter 3.3.3 --- Protein kinase C --- p.71 === Chapter 3.4 --- Induction of apoptosis by Con A --- p.89 === Chapter 3.5 --- Tubulin structure in Con A-treated cells --- p.94 === Chapter 3.6 --- Nitric oxide and Con A-induced cell death in macrophages --- p.96 === Chapter 3.7 --- Effect ofNOS inhibitor on cell death induced by Con A --- p.102 === Chapter 3.8 --- Involvement of PKC in the Con A-induced cell death in macrophages --- p.106 === Chapter Section 4 --- Discussion --- p.114 === Chapter 4.1 --- Induction of apoptosis in macrophages by NO producing drugs --- p.118 === Chapter 4.2 --- Signal transduction pathways to the NO-induced cell death in macrophages --- p.120 === Chapter 4.2.1 --- Ca2+ ion --- p.120 === Chapter 4.2.2 --- cGMP --- p.120 === Chapter 4.2.3 --- Protein kinase C --- p.121 === Chapter 4.3 --- Induction of apoptosis by Con A --- p.124 === Chapter 4.4 --- Tubulin structure in Con A-treated cells --- p.124 === Chapter 4.5 --- Nitric oxide and Con A-induced cell death --- p.125 === Chapter 4.6 --- Effect ofNOS inhibitor on cell death induced by Con A --- p.125 === Chapter 4.7 --- Involvement of PKC in the Con A-induced cell death in macrophages --- p.127 === Chapter Section 5 --- Bibliography --- p.129 === References --- p.130