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Role of cofilin, an actin cytoskeletal protein, in ischemic conditions potential therapeutic target for ischemic stroke
Table of Contents
Abstract
Acknowledgements
List of Figures
1. Introduction
1.1 Cerebral ischemia
1.2 Neuronal Cytoskeleton
1.3 Actin filaments
1.3.1 Actin structure and dynamics
1.3.2 Actin binding proteins
1.4 ADF/Cofilin family of proteins
1.4.1 Distribution of ADF/Cofilin proteins
1.4.2 Cofilin and its role in actin dynamics
1.4.3 Regulation of activity of cofilin
1.5 Cofilin activity beyond cytoskeletal regulations- Role in pathophysiological conditions of various diseases
1.6 Hypothesis and aims
2. Materials
2.1 Materials
3. Methods
3.1 Animals
3.2 pMCAO model of stroke induction
3.3 Perfusion procedure and isolation of brain
3.4 Tissue sectioning
3.5 TUNEL assay for apoptosis
3.6 Antigen retrieval
3.7 Immunohistochemistry
3.8 Protein estimation
3.9 Protein quantification using Bradford protein assay
3.10 Western blotting procedures
3.11 Cell culture
3.12 Differentiation of PC12 cells
3.13 Oxygen glucose deprivation (OGD) model of ischemia
3.14 Treatment with stressor (tertiary butyl hydro peroxide)
3.15 Immunocytochemistry
3.16 MTT cell viability assay
3.17 Statistical analysis
4. Results
4.1 Decreased expression of phosphocofilin expression in PC12 cells against t-butyl hydroperoxide induced oxidative stress
4.2 OGD induced changes in phosphocofilin expression levels
4.3 Effect of nonspecific tyrosine phosphatase inhibitor (Na3VO4) on phosphocofilin expression in OGD
4.4 Inhibitor pretreatment increases cell viability during OGD
4.5 Immunocytochemical analysis of phosphocofilin protein expression in OGD conditions
4.6 Calcineurin, a mediator involved in activation of SSH 1L (Slingshot phosphatase) contributes to the activation of cofilin during OGD conditions
4.7 Ischemia induced changes in phosphocofilin expression in mice subjected to permanent middle cerebral artery occlusion (pMCAO) model of ischemia
4.8 Immunohistochemical representation of protein changes in mice brains subjected to ischemia
5. Discussion
ReferencesBài viết liên quan
