by Josephine Wing Sze Lau. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 84-112). / Abstracts in English and Chinese. / Acknowledgment --- p.iii / Abstract --- p.iv / List of Abbreviations --- p.viii / Chapter CHAPTER ONE: --- INTRODUCTION / Chapter 1.1 --- Methamphetamine (METH) --- p.1 / Chapter 1.1.1 --- Historical Background and Epidemiology --- p.1 / Chapter 1.1.2 --- Physical Effects of METH --- p.4 / Chapter 1.1.3 --- Neurochemical Alternation of METH --- p.6 / Chapter 1.2 --- Mechanisms of METH Toxicity / Chapter 1.2.1 --- Oxidative Stress --- p.8 / Chapter 1.2.1.1 --- Superoxide (O2-) and Superoxide Dismutase (SOD) --- p.10 / Chapter 1.2.1.2 --- "Hydrogen Peroxide (H202), Catalase and Glutathione (GSH)" --- p.11 / Chapter 1.2.1.3 --- Hydroxyl Radical (OH.) --- p.12 / Chapter 1.2.1.4 --- Nitric Oxide (NO) --- p.13 / Chapter 1.2.2 --- Apoptosis --- p.16 / Chapter 1.2.3 --- Excitotoxicity --- p.17 / Chapter 1.2.4 --- Mitochondrial Dysfunction --- p.18 / Chapter 1.2.5 --- Hyperthermia --- p.21 / Chapter 1.2.5.1 --- Cyclooxygenase-2 (COX-2) --- p.23 / Chapter 1.2.5.2 --- Heme-oxygenase-1 (HO-1) --- p.25 / Chapter 1.2.5.3 --- The Effects of Nitric Oxide (NO) on COX-2 and HO-1 Expressions --- p.27 / Chapter 1.3 --- Astrocytes / Chapter 1.3.1 --- Characteristics of Astrocytes --- p.29 / Chapter 1.3.2 --- Astrocyte Functions --- p.30 / Chapter 1.3.3 --- The Role of Astrocytes in METH-induced Neurotoxicity --- p.34 / Chapter 1.4 --- Aim of Project --- p.37 / Chapter CHAPTER TWO: --- MATERIALS AND METHODS / Chapter 2.1 --- Cell Cultures / Chapter 2.1.1 --- Astrocyte Cultures --- p.42 / Chapter 2.1.2 --- CATH.a Cell line and Astrocytes Co-cultures --- p.43 / Chapter 2.2 --- Treatment / Chapter 2.2.1 --- METH Treatment --- p.44 / Chapter 2.2.2 --- Inhibition of Cyclooxygenase-2 (COX-2) and Inducible Nitric Oxide Synthase (iNOS) --- p.44 / Chapter 2.3 --- Lactate Dehydrogenase (LDH) Assay --- p.45 / Chapter 2.4 --- Assay for Reactive Oxygen Species (ROS) Formation --- p.47 / Chapter 2.5 --- Assay for Adenosine Triphosphate (ATP) Content --- p.48 / Chapter 2.6 --- Determination of Mitochondrial Membrane Potential (Δ Ψm) --- p.50 / Chapter 2.7 --- Determination of Nitrite Levels in Cultured Astrocytes --- p.51 / Chapter 2.8 --- Western Blot Analysis --- p.52 / Chapter 2.8.1 --- COX-2 --- p.53 / Chapter 2.8.2 --- HO-1 --- p.53 / Chapter 2.9 --- Viability Assay of CATH.a-Astrocyte Cocultures --- p.54 / Chapter 2.10 --- Statistics --- p.55 / Chapter CHAPTER THREE: --- RESULTS / Chapter 3.1 --- The Effects of METH Treatment on Cultured Astrocytes / Chapter 3.1.1 --- Lactate Dehydrogenase (LDH) Activities --- p.56 / Chapter 3.1.2 --- Morphological Changes --- p.56 / Chapter 3.1.3 --- The Production of Reactive Oxygen Species / Chapter 3.1.3.1 --- Rate of change (0-120 min) --- p.57 / Chapter 3.1.3.2 --- Time course (0 - 48 h) --- p.57 / Chapter 3.1.4 --- Change in ATP Content --- p.58 / Chapter 3.1.5 --- Change in Mitochondrial Membrane Potential (Δ Ψm) --- p.59 / Chapter 3.1.6 --- Nitrite levels after METH treatment / Chapter a) --- Striatal astrocytes --- p.59 / Chapter b) --- Mesencephalic astrocytes --- p.60 / Chapter c) --- Cortical astrocytes --- p.60 / Chapter 3.1.7 --- The Effects of Aminoguanidine (AG) on Nitrite Levels / Chapter a) --- Striatal astrocytes --- p.61 / Chapter b) --- Mesencephalic astrocytes --- p.62 / Chapter c) --- Cortical astrocytes --- p.62 / Chapter 3.1.8 --- The Effects of Indomethacin (INDO) on Nitrite Levels / Chapter a) --- Striatal astrocytes --- p.63 / Chapter b) --- Mesencephalic astrocytes --- p.64 / Chapter c) --- Cortical astrocytes --- p.64 / Chapter 3.1.9 --- Change in Cyclooxygenase-2 (COX-2) Protein Levels / Chapter a) --- Striatal astrocytes --- p.65 / Chapter b) --- Mesencephalic astrocytes --- p.65 / Chapter c) --- Cortical astrocytes --- p.66 / Chapter 3.1.10 --- Change in Heme-oxygenase-1 (HO-1) Protein Levels / Chapter a) --- Striatal astrocytes --- p.66 / Chapter b) --- Mesencephalic astrocytes --- p.66 / Chapter c) --- Cortical astrocytes --- p.67 / Chapter 3.2 --- Cell Viability on CATH.a-Astrocyte Cocultures After METH Treatment --- p.67 / Chapter CHAPTER FOUR: --- DISCUSSION AND CONCLUSION --- p.69 / REFERENCES --- p.84

by Wai Yuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 123-157). / Abstracts in English and Chinese. / Acknowledegment --- p.iv / Abstract --- p.v / List of Abbreviations --- p.ix / Chapter CHAPTER ONE: --- INTRODUCTION / Chapter 1.1 --- Parkinson's Disease (PD) --- p.1 / Chapter 1.1.1 --- Epidemiology --- p.1 / Chapter 1.1.2 --- Neuropathology --- p.2 / Chapter 1.1.3 --- Clinical Symptoms --- p.3 / Chapter 1.1.4 --- Treatment --- p.6 / Chapter 1.2 --- Proposed Mechanisms of Neurodegeneration in PD --- p.11 / Chapter 1.2.1 --- Oxidative Stress --- p.11 / Chapter 1.2.2 --- Mitochondrial Dysfunction --- p.13 / Chapter 1.2.3 --- Genetic Factors --- p.15 / Chapter 1.2.4 --- Environmental Factors --- p.17 / Chapter 1.2.5 --- Ageing --- p.20 / Chapter 1.3 --- "1-Methy-4-Phenyl-1,2,3,6-Tetrahydropyridine (MPTP) as a PD Model" --- p.22 / Chapter 1.3.1 --- Discovery of MPTP --- p.22 / Chapter 1.3.2 --- The Mechanisms of MPTP-induced Neurotoxicity --- p.23 / Chapter 1.4 --- Antioxidants in the Central Nervous System --- p.26 / Chapter 1.4.1 --- Superoxide Dismutase --- p.26 / Chapter 1.4.2 --- Glutathione --- p.27 / Chapter 1.5 --- Metallothioneins (MTs) --- p.29 / Chapter 1.5.1 --- Characteristics of MTs --- p.29 / Chapter 1.5.2 --- Functions of Astrocytes --- p.31 / Chapter 1.6 --- Astrocytes --- p.34 / Chapter 1.6.1 --- Characteristics of Astrocytes --- p.34 / Chapter 1.6.2 --- Functions of Astrocytes --- p.35 / Chapter 1.6.3 --- Role of Astrocytes in Parkinson's Disease --- p.39 / Chapter 1.7 --- Aim of Project --- p.41 / Chapter CHAPTER TWO: --- MATERIALS AND METHODS / Chapter 2.1 --- In Vitro Study --- p.44 / Chapter 2.1.1 --- Astrocyte Cultures --- p.44 / Chapter 2.1.2 --- Treatment Regimen --- p.46 / Chapter 2.1.2.1 --- 1 -methyl-4-phenyl-pyridinium (MPP+) Treatment --- p.46 / Chapter 2.1.2.2 --- Induction of Metallothioneins (MTs) and Glutathione (GSH) --- p.46 / Chapter 2.1.2.2.1 --- Northern Blot Analysis --- p.47 / Chapter 2.1.2.2.2 --- Immunocytochemical Staining for MTs --- p.48 / Chapter 2.1.2.2.3 --- GSH Assay --- p.49 / Chapter 2.1.2.3 --- Iron Chelation --- p.51 / Chapter 2.1.2.4 --- Combined Pretreatment --- p.51 / Chapter 2.1.3 --- Lactate Dehydrogenase (LDH) Assay --- p.51 / Chapter 2.1.4 --- "3,(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide (MTT) Assay" --- p.53 / Chapter 2.1.5 --- Reactive Oxygen Species (ROS) Assay --- p.55 / Chapter 2.1.6 --- Protein Assay --- p.56 / Chapter 2.1.7 --- Statistics --- p.57 / Chapter 2.2 --- In Vivo Study --- p.57 / Chapter 2.2.1 --- "Administration of 1 -methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)" --- p.57 / Chapter 2.2.2 --- Tyrosine Hydroxylase (TH) Immunocytochemical Staining --- p.58 / Chapter 2.2.3 --- DAT Receptor Binding Assay --- p.59 / Chapter 2.2.4 --- Dopamine (DA) and DA metabolites - High Performance Liquid Chromatography (HPLC) --- p.60 / Chapter 2.2.5 --- Statistics --- p.61 / Chapter CHAPTER THREE: --- RESULTS / Chapter 3.1 --- In Vitro Study --- p.62 / Chapter 3.1.1. --- Induction of Metallothioneins (MTs) in Astrocytes with Zinc Sulfate (ZnS04) --- p.62 / Chapter 3.1.1.1 --- Immunocytochemical changes --- p.62 / Chapter 3.1.1.2 --- Northern Blot Analysis --- p.62 / Chapter 3.1.1.3 --- The Effects of ZnSO4 Pretreatment on 1 -methyl-4-phenyl- pyridinium (MPP+)-treated Astrocytes --- p.63 / Chapter 3.1.1.3.1 --- Lactate Dehydrogenase (LDH) Activities --- p.63 / Chapter 3.1.1.3.2 --- "3,(4,5-dimethylthiazol-2-yl)2,5-diphenyl- tetrazolium bromide (MTT) Activities" --- p.67 / Chapter 3.1.1.3.3 --- Reactive Oxygen Species (ROS) Production --- p.71 / Chapter 3.1.2 --- The Effects of NAc Pretreatment on MPP+-treated Astrocytes --- p.75 / Chapter 3.1.2.1 --- Glutathione (GSH) levels --- p.75 / Chapter 3.1.2.2 --- LDH Activities --- p.77 / Chapter 3.1.2.3 --- MTT Activities --- p.80 / Chapter 3.1.2.4 --- ROS Production --- p.83 / Chapter 3.1.3 --- The Effects of Deferoxamine on MPP+-treated Astrocytes --- p.87 / Chapter 3.1.3.1 --- LDH Activities --- p.87 / Chapter 3.1.3.2 --- ROS Production --- p.89 / Chapter 3.1.4 --- The Effects of ZnSO4 and NAc Combined Treatment on MPP+-treated Astrocytes --- p.92 / Chapter 3.1.4.1 --- LDH Activities --- p.92 / Chapter 3.1.4.2 --- ROS Production --- p.95 / Chapter 3.2 --- "Effects of 1 -methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on MT-I, -II Knock-out Mice" --- p.99 / Chapter 3.2.1 --- The Effects of MPTP on Substantia Nigral (SN) Cell Loss --- p.99 / Chapter 3.2.2 --- The Effects of MPTP on Striatal (ST) and SN Dopamine Transporter (DAT) Binding --- p.99 / Chapter 3.2.3 --- The Effects of MPTP on ST Dopamine (DA) Metabolites --- p.100 / Chapter CHAPTER FOUR: --- DISCUSSION AND CONCLUSION --- p.102 / REFERENCES --- p.123