Yeung Ho Lam. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 93-107). / Abstracts in English and Chinese. / Thesis Committee --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Acknowledgments --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.x / List of Figures --- p.xiii / List of Tables --- p.xv / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Glucose Transport through the Blood Brain Barrier --- p.1 / Chapter 1.2 --- Roles of Astrocytes in the Brain --- p.4 / Chapter 1.3 --- Glucose Metabolism in Astrocytes --- p.8 / Chapter 1.4 --- Diseases Associated with Reduced Glucose Transport --- p.10 / Chapter 1.5 --- Extracellular Accumulation of Glutamate as a Cause for Epilepsy --- p.13 / Chapter 1.6 --- Regulations of Astrocyte-mediated Glutamate Uptake --- p.17 / Chapter 1.7 --- Aim and Hypothesis of the Project --- p.22 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.24 / Chapter 2.1 --- Materials --- p.24 / Chapter 2.1.1 --- Primary Rat Astrocytes --- p.24 / Chapter 2.1.2 --- Cell Culture Materials --- p.24 / Chapter 2.1.3 --- Chemicals --- p.26 / Chapter 2.1.4 --- Reagents for the Determination of Gene Expressions --- p.26 / Chapter 2.1.5 --- Reagents for the Determination of Protein Expressions --- p.29 / Chapter 2.1.6 --- Reagents for Functional Assays --- p.33 / Chapter 2.1.6.1 --- Reagents for Enzyme-Linked Immunosorbent Assay (ELISA) of IL-6 --- p.33 / Chapter 2.1.6.2 --- Reagents for Glutamate Uptake Assay --- p.33 / Chapter 2.1.6.3 --- Reagents for Extracellular Glutamate Determination Assay --- p.33 / Chapter 2.1.6.4 --- Reagents for Glucose Uptake Assay --- p.34 / Chapter 2.1.6.5 --- Reagents for MTT Assay --- p.34 / Chapter 2.1.6.6 --- Reagents for GFAP Immunostaining --- p.35 / Chapter 2.2 --- Methods --- p.36 / Chapter 2.2.1 --- Preparation of Primary Astrocytes --- p.36 / Chapter 2.2.2 --- Determination of Gene Expressions by Reverse Transcription-Polymersase Chain Reaction (RT-PCR) --- p.37 / Chapter 2.2.3 --- Determination of Protein Expressions by Western Blotting --- p.40 / Chapter 2.2.4 --- ELISA --- p.43 / Chapter 2.2.5 --- Glutamate Uptake Assay --- p.44 / Chapter 2.2.6 --- Extracellular Glutamate Determination Assay --- p.44 / Chapter 2.2.7 --- Glucose Uptake --- p.45 / Chapter 2.2.8 --- MTT Assay --- p.46 / Chapter 2.2.9 --- GFAP Immunostaining --- p.46 / Chapter 2.2.10 --- Band Intensity Quantification --- p.47 / Chapter 2.2.11 --- Statistical Analysis --- p.47 / Chapter CHAPTER 3 --- RESULTS --- p.49 / Chapter 3.1 --- Preparation of Primary Astrocyte Culture --- p.49 / Chapter 3.2 --- Effects of Glucose Deficiency on Astrocyte-mediated Glutamate Uptake --- p.51 / Chapter 3.2.1 --- Effects of Glucose Deficiency on the Expressions of Glutamate Transporters --- p.51 / Chapter 3.2.2 --- Effects of Glucose Deficiency on Glutamate Uptake in Primary Astrocytes --- p.56 / Chapter 3.3 --- Astrocytic Glucose Transport under Glucose Deficiency --- p.61 / Chapter 3.3.1 --- Effects of Glucose Deficiency on the Expressions and Secretion of Inflammatory Cytokines --- p.64 / Chapter 3.3.2 --- Effects of Exogenous Interleukin-6 on Energy Availability in Primary Astrocytes upon Glucose Deficiency --- p.70 / Chapter 3.4 --- Signaling Mechanism Mediating the Astrocytic Responses under Glucose Deficiency --- p.74 / Chapter 3.4.1 --- Effects of Glucose Deficiency on the Expressions of Total and Phosphorylated ERK1/2 in Primary Astrocytes --- p.74 / Chapter CHAPTER 4 --- DISCUSSIONS AND CONCLUSIONS --- p.81 / Chapter 4.1 --- Role of Astrocytes in Preventing Glutamate Excitotoxicity under Glucose Deficiency --- p.81 / Chapter 4.1.1 --- Neonatal Astrocytes as the Cell Model for Chronic Glucose Deficiency --- p.81 / Chapter 4.1.2 --- Effects of Glucose Deficiency on the Expressions of Glutamate Transporters and Glutamate Uptake --- p.83 / Chapter 4.1.3 --- Cytokines: Mediators for Energy Production in Astrocytes --- p.85 / Chapter 4.1.4 --- Summary of the Roles of Astrocyets under Prolonged Glucose Deficiency --- p.88 / Chapter 4.2 --- Establishment of an in vitro GlutlDS model --- p.89 / Chapter 4.3 --- Future Directions of the Project --- p.90 / Chapter 4.4 --- Conclusion --- p.92 / REFERENCES --- p.93 / APPENDIX --- p.108
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325735 |
| Date | January 2006 |
| Contributors | Yeung, Ho Lam., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xv, 123 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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