Conn The Unfolded Protein Response and Cellular Stress, Part A

1. Auflage 2011
ISBN: 978-0-12-385117-8
Verlag: Elsevier Science & Techn.
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 489, 432 Seiten, Web PDF

Reihe: Methods in Enzymology

This volume provides descriptions of the occurrence of the UPR, methods used to assess it, pharmacological tools and other methodological approaches to analyze its impact on cellular regulation. The authors explain how these methods are able to provide important biological insights. - This volume provides descriptions of the occurrence of the UPR, methods used to assess it, pharmacological tools and other methodological approaches to analyze its impact on cellular regulation - The authors explain how these methods are able to provide important biological insights

Autoren/Hrsg.

Conn, P. Michael

Inhaltsverzeichnis

1;Front Cover;1
2;Methods in Enzymology: The Unfolded Protein Response and Cellular Stress, Part A;4
3;Copyright;5
4;Contents;6
5;Contributors;12
6;Preface;18
7;Methods in Enzymology;20
8;Section 1: UPR and Cell Stress in Normal and Diseased Cells;50
8.1;Chapter 1: The Unfolded Protein Response in Mouse Cerebral Cortex;52
8.1.1;1. Introduction: The UPR;52
8.1.2;2. Transduction of the UPR Signal by PERK, IRE1, and ATF6;53
8.1.3;3. UPR and Apoptosis;54
8.1.4;4. The UPR in Health and Disease;55
8.1.5;5. Induction of the UPR by Sleep Deprivation;55
8.1.6;6. Experimental Protocols;56
8.1.7;7. Concluding Remarks;67
8.1.8;Acknowledgments;68
8.1.9;References;68
8.2;Chapter 2: Immunohistochemical Detection of the Unfolded Protein Response in Atherosclerotic Plaques;72
8.2.1;1. Endoplasmic Reticulum and the Unfolded Protein Response;73
8.2.2;2. Atherosclerosis;73
8.2.3;3. UPR in the Atherosclerotic Lesion;74
8.2.4;4. Mouse Models of Atherosclerosis;76
8.2.5;5. Methods;78
8.2.6;Acknowledgments;93
8.2.7;References;93
8.3;Chapter 3: The Role of PDI as a Survival Factor in Cardiomyocyte Ischemia;96
8.3.1;1. Introduction;97
8.3.2;2. Identification of PDI as Cardio-Protective Factor;99
8.3.3;3. Conclusions;111
8.3.4;References;111
8.4;Chapter 4: Measurement of the Increase in Endoplasmic Reticulum Stress-Related Proteins and Genes in Adipose Tissue of Obese,;116
8.4.1;1. Introduction;117
8.4.2;2. Study Subjects and Fat Biopsies;118
8.4.3;3. Proteomic Analysis;119
8.4.4;4. Western Blot Analysis;128
8.4.5;5. RT-PCR Analysis;129
8.4.6;Acknowledgments;130
8.4.7;References;131
8.5;Chapter 5: Measurement of the Unfolded Protein Response (UPR) in Monocytes;132
8.5.1;1. Introduction;133
8.5.2;2. Investigating UPR Activation in the Monocyte;135
8.5.3;Acknowledgments;142
8.5.4;References;142
8.6;Chapter 6: Assessment of Endurance Exercise Training in Hepatic Endoplasmic Reticulum Stress;146
8.6.1;1. Introduction;147
8.6.2;2. Assessment of an Endurance-Training Program;148
8.6.3;3. "Dietary-Inducible" Hepatic ER Stress;153
8.6.4;4. Assessment of Hepatic ER Stress Molecular Markers;154
8.6.5;5. Required Materials;154
8.6.6;Acknowledgments;155
8.6.7;References;155
8.7;Chapter 7: Discovery Approaches to UPR in Athero-Susceptible Endothelium In Vivo;158
8.7.1;1. Introduction;159
8.7.2;2. Procedures for the Isolation of Endothelial Cells and Preparation of RNA for Microarray Hybridization;161
8.7.3;3. Microarray Hybridization and Feature Extraction;165
8.7.4;4. Bioinformatics Analysis;166
8.7.5;5. Another Approach to Gene Connectivity: Weighted Gene Coexpression Network Analysis;171
8.7.6;6. Validation and Follow-up;173
8.7.7;Acknowledgments;174
8.7.8;References;174
8.8;Chapter 8: Assessment of Endoplasmic Reticulum Stress and the Unfolded Protein Response in Endothelial Cells;176
8.8.1;1. Introduction;177
8.8.2;2. The Acute Phase of ER Stress;178
8.8.3;3. The Adaptive Reaction Phase: Unfolded Protein Response in Endothelial Cells;180
8.8.4;4. The Final Phase: Cell Death Signaling;189
8.8.5;Acknowledgments;194
8.8.6;References;194
8.9;Chapter 9: Measuring the Impact of Cigarette Smoke on the UPR;196
8.9.1;1. Introduction;197
8.9.2;2. Preparing Samples for Analysis;198
8.9.3;3. Analysis of Activation of the PERK Pathway;204
8.9.4;4. ATF6 Pathway;207
8.9.5;5. IRE1 Pathway;208
8.9.6;6. Using LSC to Measure UPR Activation;208
8.9.7;Acknowledgments;210
8.9.8;References;211
8.10;Chapter 10: Induction and Measurement of UPR and Osmotic Stress in the Yeast Pichia pastoris;214
8.10.1;1. Introduction;215
8.10.2;2. Measuring UPR Induction in P. pastoris;218
8.10.3;3. Inducing and Measuring Osmotic Stress in P. pastoris;228
8.10.4;Acknowledgments;235
8.10.5;References;235
8.11;Chapter 11: Induction of ER Stress Response Leading to Programmed Cell Death in Trypanosoma brucei;238
8.11.1;1. Introduction;239
8.11.2;2. Induction of ER Stress in Trypanosomes;242
8.11.3;3. Measurements of ER Stress in Trypanosomes Cells;243
8.11.4;4. Measurements of ER Stress-Induced Cell Death;250
8.11.5;Acknowledgments;252
8.11.6;References;252
8.12;Chapter 12: Measurement of Activation of the Endoplasmic Reticulum Stress Response in Autoimmune Myositis;256
8.12.1;1. Introduction;257
8.12.2;2. Measuring Endoplasmic Reticulum Stress Response in Skeletal Muscle Tissues Using Western Blotting;258
8.12.3;3. Immunohistochemical Detection of ER Stress Markers in Muscle Tissues;265
8.12.4;4. Determination of mRNA Levels of ER Stress Targets;270
8.12.5;5. Concluding Remarks;272
8.12.6;Acknowledgments;272
8.12.7;References;272
8.13;Chapter 13: Activation of the Unfolded Protein Response in Human Acute Myeloid Leukemia;276
8.13.1;1. Introduction;277
8.13.2;2. Technical Aspects;281
8.13.3;3. Discussion;288
8.13.4;References;290
8.14;Chapter 14: Modulation of the Unfolded Protein Response by GRP78 in Prostate Cancer;294
8.14.1;1. Introduction;295
8.14.2;2. Methods for Monitoring UPR Modulation;297
8.14.3;3. Conclusions;305
8.14.4;References;305
8.15;Chapter 15: Endoplasmic Reticulum Stress in Brain Damage;308
8.15.1;1. Introduction;309
8.15.2;2. ER Stress and Unfolded Protein Response;310
8.15.3;3. Cross Talk Between ER and Mitochondria;316
8.15.4;4. Experimental Approaches for the Detection of ER Stress;318
8.15.5;Acknowledgments;320
8.15.6;References;321
8.16;Chapter 16: Methods and Models for Monitoring UPR-Associated Macrophage Death During Advanced Atherosclerosis;326
8.16.1;1. Introduction to ER Stress in Atherosclerosis;327
8.16.2;2. Significance of Macrophage ER Stress and Apoptosis in Advanced Atherosclerosis;328
8.16.3;3. Modeling Atherosclerosis-Relevant ER Stress-Induced Apoptosis;330
8.16.4;4. The UPR and Macrophage Death at the Murine Aortic Root;334
8.16.5;5. Conclusion and Future Methodological Advances;342
8.16.6;Acknowledgments;342
8.16.7;References;342
8.17;Chapter 17: Detecting Autophagy in Response to ER Stress Signals in Cancer;346
8.17.1;1. Introduction;347
8.17.2;2. Monitoring Autophagy Upon ER Stress;348
8.17.3;3. Monitoring Autophagy Upon ER Stress In Vivo;358
8.17.4;References;366
8.18;Chapter 18: The Unfolded Protein Response Induced by Salt Stress in Arabidopsis;368
8.18.1;1. Introduction;368
8.18.2;2. The Treatment of Arabidopsis Seedlings;369
8.18.3;3. The Analysis of Unfolded Protein Response;371
8.18.4;Acknowledgments;376
8.18.5;References;376
8.19;Chapter 19: Measurement of ER Stress Response and Inflammation in the Mouse Model of Nonalcoholic Fatty Liver Disease;378
8.19.1;1. Introduction;379
8.19.2;2. Methods to Measure ER Stress Response and Hepatic Inflammation in the Mouse Model of NAFLD;381
8.19.3;3. Visualizing ER Stress Response Associated with Liver Pathology in the Mouse Model of NAFLD;390
8.19.4;Acknowledgments;394
8.19.5;References;394
8.20;Author Index;398
8.21;Subject Index;414
8.22;Colour Plate;420