Part I. Introduction to the Renin-Angiotensin System Historical Perspective of the Renin-Angiotensin System John E. Hall Three Angiotensin Paradigms in One Patient: Etiology of Hypertension, Glomerular Hemodynamics, and Long-Term Glomerular Protection Cheryl L. Laffer and Fernando Elijovich Part II. Transgenic and Gene Targeting Methods Production and Use of Chimeric Mice Junji Takaya, Taiji Matsusaka, and Iekuni Ichikawa Genetic Manipulation of the Renin-Angiotensin System Using Cre-loxP-Recombinase Curt D. Sigmund and David E. Stec Systemic Delivery of a Transgene in Intact Animals by a Retroviral Vector Michael J. Katovich, Hong-Wei Wang, Craig H. Gelband, and Mohan K. Raizada Antisense Inhibition of the Renin-Angiotensin System Dagmara Mohuczy and M. Ian Phillips Part III. Analysis of Transcriptional and Translational Mechanisms Analysis of Transcriptional Control Mechanisms I: Techniques for Characterization of cis-Regulatory Elements Christopher T. Sherman and Allan R. Brasier Analysis of Transcriptional Control Mechanisms II: Techniques for Characterization of trans-Acting Factors Allan R. Brasier, Christopher T. Sherman, and Mohammad Jamaluddin Analysis of Cytosolic Proteins that Bind to the 5' Leader Sequence of the Angiotensin AT1 Receptor by RNA Electromobility Shift Assay Zheng Wu, Kamakshi Krishnamurthi, Koby Mok, and Kathryn Sandberg In Vitro Translation of the Angiotensin AT1 Receptors in Wheat-Germ Extracts Hong Ji, Kamakshi Krishnamurthi, Zheng Wu, and Kathryn Sandberg Part IV. Assays for Determination of mRNA Levels Quantitation of Angiotensin II Receptors by Competitive Reverse-Transcription Polymerase Chain Reaction Wei Wu, Aqing Yao, and Donna H. Wang Analysis of RNA by Northern-Blot Hybridization Patricia E. Gallagher and Debra I. Diz Use of the Ribonuclease Protection Assay for the Analysis and Characterization of Target mRNAs Scott C. Supowit and Donald J. DiPette In Situ Hybridization Susan Riggs Runge, Zhaoyong Hu, and Marschall Runge Part V. Assays for Determination of Protein Contents and Enzyme Activity Immunohistochemical Detection of Angiotensin II Receptor Zhi-Qin Wang and Robert M. Carey Western Blotting Nageswara R. Madamanchi and Marschall S. Runge Quantification of Angiotensin-Converting Enzyme (ACE) Activity Qing Cheng Meng and Kathleen H. Berecek Quantification of Angiotensin Peptides Qing Cheng Meng and Kathleen H. Berecek Radiolabeling of Angiotensin Peptides Robert C. Speth and Joseph W. Harding Assays for Radiolabel-Photoaffinity Labeling of Angiotensin Receptors Jacqueline Perodin, Antony A. Boucard, Richard Leduc, Emanuel Escher, and Gaetan Guillemette Radioligand Binding Assay Gaetan Thibault and Ernesto L. Schiffrin Autoradiographic Localization and Quantification of Components of the Renin-Angiotensin System in Tissues Siew Yeen Chai, Andrew M. Allen, Jialong Zhuo, Ingrid Moeller, and Frederick A. O. Mendelsohn Part VI. Biochemical Studies of the Renin-Angiotensin System Measurement of Intracellular Free Calcium Ion Concentration in Vascular Smooth Muscle Cells: Fluorescence Imaging of Cytosolic Calcium Rhian M. Touyz and Ernesto L. Schiffrin Measurement of Phospholipidase D Activation in Vascular Smooth Muscle Cells Rhian M. Touyz and Ernesto L. Schiffrin Measurement of Cyclic AMP and Cyclic GMP in Xenopus Oocytes Stimulated with Angiotensin II and Atrial Natriuretic Factor Hong Ji Measurement of Nitric Oxide Formation Christian Thorup and Leon C. Moore Measuring Renin Secretion from Juxtaglomerular