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Biochemical and genetic analysis of Tau protein kinases in drosophila. / Biochemical & genetic analysis of Tau protein kinases in drosophila

Chau Wing-Kam. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 92-101). / Abstracts in English and Chinese. / Abstract --- p.I / Abstract (Chinese version) --- p.III / Acknowledgement --- p.IV / List of Abbreviations --- p.VIII / List of Tables --- p.IX / List of Figures --- p.X / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Neurodegenerative diseases --- p.2 / Chapter 1.2 --- Tauopathies --- p.5 / Chapter 1.3 --- Function and structure of Tau --- p.9 / Chapter 1.4 --- Post-translational modifications of Tau --- p.13 / Chapter 1.5 --- Tau protein kinases --- p.17 / Chapter 1.6 --- Tau protein kinase inhibitors --- p.19 / Chapter 1.7 --- Drosophila model of Tauopathies --- p.20 / Chapter 1.8 --- Aims of study --- p.24 / Chapter Chapter 2 --- Materials and methods / Chapter 2.1 --- Drosophila manipulation / Chapter 2.1.1 --- Drosophila genetics --- p.26 / Chapter 2.1.2 --- External fly eye and adult wing morphology examination --- p.27 / Chapter 2.1.3 --- Study of fly wings deformation on Tau kinase overexpression --- p.27 / Chapter 2.2 --- RNA extraction / Chapter 2.2.1 --- Method --- p.28 / Chapter 2.2.2 --- Buffers and reagents --- p.29 / Chapter 2.3 --- Reverse transcription-PCR / Chapter 2.3.1 --- Method --- p.30 / Chapter 2.3.2 --- Buffers and reagents --- p.31 / Chapter 2.4 --- SDS-Polyacrylamide gel electrophoresis / Chapter 2.4.1 --- Method --- p.31 / Chapter 2.4.2 --- Buffers and reagents --- p.32 / Chapter 2.5 --- Western blotting / Chapter 2.5.1 --- Method --- p.32 / Chapter 2.5.2 --- Buffers and reagents --- p.33 / Chapter 2.6 --- Phosphatase treatment of proteins / Chapter 2.6.1 --- Method --- p.34 / Chapter 2.6.2 --- Buffers and reagents --- p.34 / Chapter 2.7 --- Sequential extraction of proteins / Chapter 2.7.1 --- Methods --- p.35 / Chapter 2.7.2 --- Buffers and reagents --- p.36 / Chapter 2.8 --- Sarkosyl extraction of proteins / Chapter 2.8.1 --- Method --- p.37 / Chapter 2.8.2 --- Buffers and reagents --- p.37 / Chapter 2.9 --- Immunostaining / Chapter 2.9.1 --- Method --- p.38 / Chapter 2.9.2 --- Buffers and reagents --- p.38 / Chapter 2.10 --- Lithium treatment of flies / Chapter 2.10.1 --- Method --- p.39 / Chapter 2.10.2 --- Buffers and reagents --- p.40 / Chapter 2.11 --- Quantitation of Lithium ion by atomic absorption spectrometry / Chapter 2.11.1 --- Method --- p.40 / Chapter 2.12 --- Statistical analysis --- p.41 / Chapter Chapter 3 --- Results / Chapter 3.1 --- GAL4/UAS gene expression system in transgenic fly / Chapter 3.1.1 --- Introduction --- p.43 / Chapter 3.1.2 --- Results --- p.47 / Chapter 3.1.3 --- Discussion --- p.52 / Chapter 3.2 --- Tau phosphorylation and Tau-induced toxicity in transgenic fly / Chapter 3.2.1 --- Introduction --- p.52 / Chapter 3.2.2 --- Results / Chapter 3.2.2.1 --- Overexpressed Tau is phosphorylated and toxic in fly --- p.53 / Chapter 3.2.2.2 --- Coexpression of GSK3β/Shaggy or Cdk5 enhance Tau phosphorylation and Tau-induced toxicity --- p.57 / Chapter 3.2.2.3 --- Lithium suppresses Tau phosphorylation and Tau-induced toxicity --- p.64 / Chapter 3.2.3 --- Discussion --- p.68 / Chapter 3.3 --- Tau solubility properties in transgenic fly / Chapter 3.3.1 --- Introduction --- p.69 / Chapter 3.3.2 --- Results / Chapter 3.3.2.1 --- Coexpression of GSKlβ/Shaggy does not alter the sarkosyl solubility of Tau --- p.70 / Chapter 3.3.2.2 --- Coexpression of GSK3β/Shaggy causes a minor alteration of Tau solubility properties --- p.73 / Chapter 3.3.3 --- Discussion --- p.78 / Chapter 3.4 --- Tau aggregate formation in transgenic fly / Chapter 3.4.1 --- Introduction --- p.79 / Chapter 3.4.2 --- Results / Chapter 3.4.2.1 --- Tau aggregates are detected in aged transgenic flies --- p.80 / Chapter 3.4.3 --- Discussion --- p.82 / Chapter 3.5 --- Effect of Lithium on GSK3p/Shaggy-induced wing deformation / Chapter 3.5.1 --- Introduction --- p.83 / Chapter 3.5.2 --- Results / Chapter 3.5.2.1 --- Lithium rescues GSK3β/Shaggy-induced wing deformation --- p.84 / Chapter 3.5.3 --- Discussion --- p.86 / Chapter Chapter 4 --- General discussion --- p.87 / References --- p.92

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325122
Date January 2005
ContributorsChau, Wing-Kam., Chinese University of Hong Kong Graduate School. Division of Molecular Biotechnology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, x, 101 leaves : ill. (some col.) ; 30 cm.
RightsUse 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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