A6222
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Angiotensin Protocols

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General description

The renin-angiotensin system has been in the spotlight fo rmore than 100 years because of its vital role in blood pressure regulation, ion and fluid homeostasis, and tissue growth and remodeling. This book describes in step-by-step detail all the major molecular, biochemical, and functional techniques used in probing the renin-angiotensin system. These range from the use of embryonic gene targeting to create of mutant strains, to the functional investigation of components of the renin-angiotensin system. This comprehensive text translates expert knowledge into readily reproducible methods that enable all biomedical investigators successfully to explore the role of angiotensin in hypertension, congestive heart failure, chronic renal failure and stroke.

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Identifiers

ISBN-10

0-89603-763-0

ISBN-13

978-0-89603-763-2

Properties

publication info

D.H. Wang, ed., Humana Press, 2001, 544 pp., hard cover

Table of Contents

PART I INTRODUCTION TO THE RENIN-ANGIOTENSIN SYSTEM
1. Historical Perspective of the Renin-Angiotensin System
2. Three Angiotensin Paradigms in One Patient: Etiology of Hypertension, Glomerular Hemodynamics, and LongTerm Glomerular Protection
PART II TRANSGENIC AND GENE TARGETING METHODS
3. Production and Use of Chimeric Mice
4. Genetic Manipulation of the Renin-Angiotensin System Using Cre-loxP-Recombinase
5. Systemic Delivery of a Transgene in Intact Animals by a Retroviral Vector
6. Antisense Inhibition of the Renin-Angiotensin System
PART III ANALYSIS OF TRANSCRIPTIONAL AND TRANLATIONAL MECHANISMS
7. Analysis of Transcriptional Control Mechanisms I: Techniques for Characterization of cis-Regulatory Elements
8. Analysis of Transcriptional Control Mechanisms II: Techniques for Characterization of trans-Acting Factors
9. Analysis of Cytosolic Proteins that Bind to the 5' Leader Sequence of the Angiotensin AT₁ Receptors by RNA Electromobility Shift Assay
10. In Vitro Translation of the Angiotensin AT₁ Receptors in Wheat Germ Extracts
PART IV ASSAYS FOR DETERMIANTION OF mRNA LEVELS
11. Quantitation of Angiotensin II Receptors by Competitive Reverse Transcription Polymerase Chain Reaction
12. Analysis of RNA by Northern Blot Hybridization
13. Use of the Ribonuclease Protection Assay for the Analysis and Charactirzation of Target mRNAs
14. In Situ Hybridization
PART V ASSAYS FOR DETERMINATION OF PROTEIN CONTENTS AND ENZYME ACTIVITY
15. Immunohistochemical Detection of Angiotensin II Receptor
16. Western Blotting
17. Quantification of Angiotensin Converting Enzyme (ACE) Activity
18. Quantification of Angiotensin peptides
19. Radiolabeling of Angiotensin Peptides
20. Assyas fo rRadiolabel Photoaffinity Labeling of Angiotensin Recetpors
21. Radioligand Binding Assay
22. Autoradiographic Localization and Quantifications of Components of the Renin-Angiotensin System in Tissues
PART VI BIOCHEMICAL STUDIES OF THE RENIN-ANGIOTNSIN SYSTEM
23. Measurement of Intracellular Free Calcium Ion Concentration in Vascular Smooth Muscle Cells: Fluorescence Imaging of Cytosolic Calcium
24. Measurement of Phospholipase D Activation in Vascular Smooth Muscle Cells
25. Measurement of Cyclic AMP and Cyclic GMP in Xenopus Oocytes Stimulated with Angiotensin II and Atrial Natriuretic Factor
28. Measurement of Nitric Oxide Formation
27. Mesuring Renin Secretion form Juxtaglomerular Cell
PART VI FUNCTIONAL STUDIES OF TH ERENIN-ANGIOTENSIN SYSTME
28. Measurement of Regional Blood Flow int eh Kidney Using Laser-Doppler Flowmetry
29. Measurement of Renal Tubular Angiotensin II

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