BCL-2 family in retinal degeneration in ischemia/reperfusion injury and in the RCS rats.

Chiu Kin.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1998.
Includes bibliographical references (leaves 100-116).
also in Chinese.
TABLE OF CONTENTS --- p.I
ACKNOWLEDGEMENTS --- p.V
LIST OF FIGURES --- p.VI
LIST OF ABBREVIATIONS --- p.VIII
p.1
Chapter 1. --- INTRODUCTION --- p.5
Chapter 2. --- LITERATURE REVIEW --- p.7
Chapter 2.1 --- RETINAL ISCHEMIA --- p.7
Chapter 2.1.1 --- INDUCTION OF RETINAL ISCHEMIA --- p.7
Chapter 2.1.2 --- MECHANISMS OF RETINAL ISCHEMIA/REPERFUSION DAMAGE --- p.8
Chapter 2.1.2.1. --- Free radical --- p.8
Chapter 2.1.2.2 --- Excitotoxicity --- p.9
Chapter 2.1.3 --- APOPTOSIS IN RETINAL ISCHEMIA/REPERFUSION INJURY --- p.10
Chapter 2.2 --- RETINAL DYSTROPHIC ROYAL COLLEGE OF SURGEONS (RCS) RAT --- p.15
Chapter 2.3 --- BCL-2 FAMILY MEMBERS --- p.16
Chapter 2.3.1 --- FAMILY MEMBERS AND THEIR INTERACTIONS --- p.16
Chapter 2.3.2 --- SUBCELLULAR LOCALIZATION --- p.18
Chapter 2.3.3 --- PHYSICAL STRUCTURE AND PORE FORMATION --- p.19
Chapter 2.3.4 --- BIOLOGICAL EFFECTS OF BCL-2 --- p.20
Chapter 3. --- OBJECTIVES --- p.24
Chapter 4. --- MATERIALS AND METHODS --- p.27
Chapter 4.1. --- RETINAL ISCHEMIA AND REPERFUSION INDUCED LOSS OF INNER RETINAL ELEMENTS --- p.27
Chapter 4.1.1. --- TISSUE RESPONSES IN THE RAT RETINAS AFTER TRANSIENT ELEVATED INTRAOCULAR PRESSURE INDUCED RETINAL ISCHEMIA/REPERFUSION INSULT --- p.27
Chapter 4.1.1.1. --- Induction of retinal ischemia/reperfusion insult with transient elevated intraocular pressure (IOP) --- p.27
Chapter 4.1.1.2. --- Animal experiments --- p.28
Chapter 4.1.1.3. --- Histopathology and measurement of inner retinal thickness (IRT) --- p.28
Chapter 4.1.1.4. --- Flat preparation of the retinas and retinal ganglion cell counts (RGCCs) --- p.29
Chapter 4.1.2. --- INTERNUCLEOSOMAL DNA FRAGMENTATION AND IN SITU NICKED DNA DETECTIONS AT DIFFERENT TIME AFTER REPERFUSION IN THE RAT RETINAS --- p.30
Chapter 4.1.2.1. --- Enzyme-linked immunosorbent assay (ELISA) of mono- and oligonucleosomes --- p.30
Chapter 4.1.2.2. --- In-situ terminal deoxynucleotidyl transferase (TdT)-mediated biotin- dUTP nicked end labelling (TUNEL) --- p.31
Chapter 4.1.3. --- "IMMUNOHISTOCHEMISTRY OF BCL-2, BAX AND P53" --- p.32
Chapter 4.1.4. --- "DOUBLE LABELLING OF BCL-2, BAX AND TUNEL" --- p.33
Chapter 4.1.5. --- IN-SITU REVERSE TRANSCRIPTASE - POLYMERASE CHAIN REACTION OF BCL-2 AND BAX --- p.34
Chapter 4.1.5.1. --- Primers design and specificity test --- p.34
Chapter 4.1.5.2. --- In-situ RT-PCR --- p.36
Chapter 4.2. --- LOSS OF INNER RETINAL ELEMENTS IN THE RETINAL DYSTROPHIC ROYAL COLLEGE OF SURGEONS (RCS) RATS --- p.38
Chapter 4.2.1. --- HISTOPATHOLOGY --- p.38
Chapter 4.2.2 --- MORPHOMETRY OF CELLS IN THE RETINAL GANGLION CELL LAYER (RGCL) AND THE INNER NUCLEAR LAYER (INL) --- p.39
Chapter 4.2.3. --- IMMUNOHISTOCHEMISTRY OF BCL-2 AND BAX --- p.39
Chapter 5. --- RESULTS --- p.40
Chapter 5.1. --- RETINAL ISCHEMIA AND REPERFUSION INDUCED LOSS OF INNER RETINAL ELEMENTS --- p.40
Chapter 5.1.1. --- TISSUE RESPONSES IN THE RAT RETINAS AFTER TRANSIENT ELEVATED INTRAOCULAR PRESSURE INDUCED ISCHEMIA/ REPERFUSION INSULT --- p.40
Chapter 5.1.1.1. --- Histopathology --- p.40
Chapter 5.1.1.2. --- Morphometry of inner retinal thickness --- p.40
Chapter 5.1.1.3. --- Retinal ganglion cell counts (RGCCs) --- p.41
Chapter 5.1.2. --- INTERNUCLEOSOMAL DNA FRAGMENTATION AND IN SITU NICKED DNA DETECTION AT DIFFERENT TIME AFTER REPERFUSION IN THE RAT RETINAS --- p.41
Chapter 5.1.2.1. --- Enzyme-linked immunosorbent assay (ELISA) of mono- and oligonucleosomes --- p.42
Chapter 5.1.2.2. --- In situ TUNEL --- p.42
Chapter 5.1.3. --- BCL-2 AND RETINAL ISCHEMIA/REPERFUSION INJURY --- p.42
Chapter 5.1.3.1. --- Immunohistochemistry of Bcl-2 --- p.42
Chapter 5.1.3.2. --- Double labelling of Bcl-2 and TUNEL --- p.43
Chapter 5.1.3.3. --- In situ RT-PCR for bcl-2 mRNA --- p.43
Chapter 5.1.4. --- BAX AND RETINAL ISCHEMIA/REPERFUSION INJURY --- p.44
Chapter 5.1.4.1. --- Immunohistochemistry of Bax --- p.44
Chapter 5.1.4.2. --- Double labelling of Bax and TUNEL --- p.45
Chapter 5.1.4.3. --- In situ RT-PCR for bax mRNA --- p.45
Chapter 5.1.5. --- P53 IMMUNOREACTIVITY AT VARIOUS TIME AFTER REPERFUSION --- p.46
Chapter 5.2. --- LOSS OF INNER RETINAL ELEMENTS IN THE RETINAL DYSTROPHIC ROYAL COLLEGE OF SURGEON (RCS) RATS --- p.47
Chapter 5.2.1. --- HISTOPATHOLOGY --- p.47
Chapter 5.2.2. --- MORPHOMETRY OF CELLS IN THE RGCL AND INL --- p.47
Chapter 5.2.3. --- IMMUNOHISTOCHEMISTRY OF BCL-2 AND BAX --- p.47
Chapter 5.2.3.1. --- Bcl-2 --- p.47
Chapter 5.2.3.2. --- Bax --- p.48
Chapter 6. --- DISCUSSION --- p.49
Chapter 6.1. --- RETINA ISCHEMIA AND REPERFUSION INDUCED LOSS OF RETINAL ELEMENTS --- p.51
Chapter 6.1.1 --- REPERFUSION TIME DEPENDENT TISSUE RESPONSES IN RAT RETINAS --- p.51
Chapter 6.1.2 --- ISCHEMIA/REPERFUSION INDUCED APOPTOSIS IN RAT RETINAS --- p.52
Chapter 6.1.3 --- BCL-2 AND RETINAL ISCHEMIA/REPERFUSION INSULT --- p.53
Chapter 6.1.4 --- BAX AND RETINAL ISCHEMIA/REPERFUSION INSULT --- p.58
Chapter 6.1.5 --- P53 AND RETINAL ISCHEMIA/REPERFUSION INJURY --- p.60
Chapter 6.2. --- LOSS OF INNER RETINAL ELEMENTS IN THE RETINAL DYSTROPHIC ROYAL COLLEGE OF SURGEON (RCS) RAT --- p.61
Chapter 6.2.1. --- HISTOPATHOLOGY AND MORPHOMETRY --- p.62
Chapter 6.2.2. --- BCL-2 --- p.63
Chapter 6.2.3. --- BAX --- p.64
Chapter 7. --- CONCLUSION --- p.65
APPENDIX A FIGURES --- p.66
APPENDIX B REFERENCES --- p.100
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