A biochemical study of cell death in murine PU5-1.8 cells.

by Chan Chun-wai, Francis.Thesis (M.Phil.)--Chinese University of Hong Kong, 1993.Includes bibliographical references (leaves 105-116).Abstract --- p.IAcknowledgments --- p.IIIAbbreviations --- p.IVObjectives --- p.VIContent --- p.VIIChapter Section 1 --- IntroductionChapter I. --- Preamble --- p.1Chapter II. --- Characteristics of Cell Death Process --- p.1Chapter II.1. --- Necrosis --- p.1Chapter II.2. --- Apoptosis-Programmed Cell Death --- p.5Chapter III. --- Triggering of Programmed Cell Death --- p.10Chapter IV. --- DNA Fragmentation and Activation of Endogenous Endonuclease --- p.12Chapter V. --- Signal Transduction Leading to Programmed Cell Death --- p.14Chapter V.1. --- Role of Calcium Ion --- p.14Chapter V.2. --- Role of Protein Kinase C --- p.15Chapter V.3. --- Protein Dephosphorylation by Phosphatases --- p.16Chapter V.4. --- Role of Adenosine 3':5'-cyclic Monophosphate --- p.17Chapter V.5. --- Other Signaling Mechanisms --- p.17Chapter VI. --- Gene Regulation in Programmed Cell Death --- p.19Chapter VI. 1. --- Gene Expression in Programmed cell death --- p.19Chapter VI. 1.1 . --- Tissue Transglutaminase --- p.19Chapter VI. 1.2. --- Poly (ADP-ribose) Polymerase --- p.20Chapter VI. 1.3. --- Testosterone-Repressed Prostate Message-2 Gene --- p.20Chapter VI. 1.4. --- Other Programmed Cell Death Associated Gene Expressions --- p.21Chapter VI.2. --- Protooncogene Regulation in Programmed Cell Death --- p.22Chapter VI.2.1. --- bcl-2 Expression --- p.22Chapter VI.2.2. --- c-myc Expression --- p.23Chapter VII. --- Concanavalin A and succinylated Concanavalin A --- p.25Chapter VII. 1. --- Physiochemical Characterization --- p.25Chapter VII.2. --- Cellular Response to Concanavalin A --- p.29Chapter VIII. --- Features of Murine Macrophage Cell Line PU5-1.8 and Normal Macrophages --- p.32Chapter Section 2 --- Materials and MethodsChapter I. --- Materials --- p.33Chapter II. --- Cell Culture --- p.33Chapter III. --- [Methyl-3H]-Thymidine Incorporation Assay --- p.34Chapter IV. --- [Methyl-3H]-Thymidine Release Assay --- p.34Chapter V. --- "3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT ) Cell Death Assay" --- p.35Chapter VI. --- Identification of Cell Death using DNA Chelating Fluorescence Probes´ؤFluorescent Microscopy and Confocal Laser Microscopy --- p.35Chapter VII. --- Analysis of DNA Fragmentation --- p.37Chapter VIII. --- Determination of Fluxes by Confocal Laser Microscopy --- p.38Chapter IX. --- Determination of PKC Activation by Western Blotting and Immunocytochemistry --- p.39Chapter X. --- Statistical Analysis --- p.41Chapter Section 3 --- ResultsChapter I. --- Concanavalin A was a Cell Death Causing Agent in PU5-1.8 cells --- p.42Chapter I.1 --- Con A Reduced the Cell Proliferation in PU5-1.8 cells --- p.42Chapter I.2. --- Con A Exhibited Cytotoxic Effect to PU5-1.8 cells --- p.44Chapter I.3. --- Con A Exhibited Cytotoxic Effect on Normal Peritoneal Macrophages --- p.46Chapter I.4. --- Succinylated Concanavalin A Showed a Weaker Cytotoxic Effect in the PU5-1.8 cells --- p.46Chapter I.5. --- α-D-Methylmannopyranoside Inhibited the Cytotoxic Effect of Con A in PU5-1.8 cells --- p.50Chapter I.6. --- FCS Inhibited the Con A-induced cell death of PU5-1.8 cells --- p.52Chapter II. --- Concanavalin A was an Apoptosis Causing Agentin PU5-1.8 cells --- p.57Chapter II. 1. --- Con A Induced Apoptosis in PU5-1.8 cells --- p.57Chapter II. 2. --- Con A Enhanced the Release of DNA in PU5-1.8 cell --- p.63Chapter II. 3. --- Con A Induced DNA fragmentation in PU5-1.8 cells --- p.63Chapter II.4. --- Cycloheximide Inhibited the Con A-Induced Cell Death in PU5-1.8 cells --- p.67Chapter II.5. --- Nicotinamide Inhibited the Con A-Induced Cell Death in PU5-1.8 cells --- p.71Chapter III. --- Signaling elicited by Concanavalin A --- p.74Chapter III.1. --- Con A Increased Intracellular Free Calcium Ion Concentration of PU5-1.8 cells --- p.74Chapter III. 1.1. --- Con A Induced Ca2+ Mobilization in PU5-1.8 cells --- p.74Chapter III. 1.2. --- Con A Induced the Ca2+ Influx and Intracellular Ca2+ Mobilization --- p.78Chapter III. 1.3. --- BAPTA-AM Inhibited the Ca2+ Mobilization in PU5-1.8 cells Stimulated by Con A --- p.80Chapter III.2. --- Role of Protein kinase C --- p.86Chapter III.2.1. --- Con A Increased the amount of PKC in PU5-1.8 cells --- p.86Chapter III.2.2. --- Con A translocated the Protein Kinase C from Cytosol into Subnuclear Region --- p.86Chapter III.2.3. --- The Cell Death Induced by Con A Is Partially Inhibited by PKC Depletion But not by Staurosporine --- p.89Chapter Section 4 --- DiscussionsChapter I. --- PU5-1.8 cells as a Model for the Study of Cell Deathin Macrophages --- p.94Chapter II. --- Concanavalin A caused Cell Death in PU5-1.8 cells --- p.95Chapter III. --- Concanavalin A induced Programmed Cell Death in PU5-1.8 cells --- p.97Chapter IV. --- Increase in Intracellular Calcium was not Required in Con A-induced Cell Death --- p.100Chapter V. --- Activation of Protein Kinase C was Partially Required for Con A-induced Cell Death --- p.101Chapter VI. --- General Discussions --- p.102Chapter Section 5 --- Bibliography --- p.104Reference --- p.10