Abstract
The renin-angiotensin system (RAS) is a complex circulating and tissue-based system. There are multiple pathways for the formation and degradation of peptides. In order to understand the functions of the system, characterization of angiotensin peptides (products and substrates) is important. Radioimmunoassays with the requisite specificity and sensitivity have been developed to allow for the characterization and quantification of circulating and tissue angiotensins. Here, we describe the appropriate methods for collecting the tissue and blood, the extractions steps required to partially purify and remove larger molecular weight-interfering proteins from tissue and plasma, and the radioimmunoassay of three of the peptides of this system (Ang I, Ang II, and Ang-(1-7)), as well as the verification of immunoreactive identity for Ang II and Ang-(1-7) by combined high-performance liquid chromatography—RIA analysis.
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References
Chappell MC, Brosnihan KB, Diz DI, Ferrario CM (1989) Identification of angiotensin-(1-7) in rat brain. Evidence for differential processing of angiotensin peptides. J Biol Chem 264:16518–16523
Kohara K, Tabuchi Y, Senanayake P, Brosnihan KB, Ferrario CM (1991) Reassessment of plasma angiotensins measurement: effects of protease inhibitors and sample handling. Peptides 12:1135–1141
Campbell DJ, Kladis A, Valentijn AJ (1995) Effects of losartan on angiotensin and bradykinin peptides and angiotensin-converting enzyme. J Cardio Pharm 26:233–240
Sampaio WO, Nascimento AAS, Santos RA (2003) Regulation of cardiovascular signaling by kinins and products of similar converting enzyme systems. Systemic and regional hemodynamic effects of angiotensin-(1-7) in rats. Am J Physiol Heart Circ Physiol 284:H1985–H1994
Tallant EA, Clark MA (2003) Molecular mechanisms of inhibition of vascular growth by angiotensin-(1-7). Hypertension 42:574–579
Machado RD, Santos RA, Andrade SP (2000) Opposing actions of angiotensins on angiogenesis. Life Sci 66:67–76
Capettini LS, Montecucco R, Mach R, Stergiopulos N, Santos RA, da Silva RF (2012) Role of renin-angiotensin system in inflammation, immunity and aging. Curr Pharm Des 18:963–970
Chappell MC (2016) Biochemical evaluation of renin-angiotensin system: the good, bad and absolute? Am J Physiol Heart Circ Physiol 310:H137–H152
Modrall JG, Sadjadi J, Brosnihan KB, Gallagher PE, Yu CH, Bernstein KE, Chappell MC (2004) Depletion of tissue angiotensin-converting enzyme differentially influences the intrarenal and urinary expression of angiotensins. Hypertension 43:849–853
Kohara K, Brosnihan KB, Ferrario CM (1993) Angiotensin-(1-7) in the spontaneously hypertensive rat. Peptides 14:883–891
Westwood BM, Chappell MC (2012) Differential metabolism of angiotensin-(1-12) in the circulation and kidney of the mRen2.Lewis congenic rat. Peptides 35:190–195
Yamaleyeva L, Gilliam-Davis S, Almeida I, Brosnihan KB, Lindsey SH, Chappell MC (2012) Differential regulation of circulating and renal ACE2 and ACE in hypertensive mRen2.Lewis with early onset diabetes. Am J Physiol Renal Physiol 302:F1374–F1384
Acknowledgment
Support was provided in part by the National Institutes of Health, NHLBI-P01 HL51952, HL56932, HD084227, Unifi, Inc. Greensboro, NC and Farley-Hudson Foundation, Jacksonville, NC.
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Brosnihan, K.B., Chappell, M.C. (2017). Measurement of Angiotensin Peptides: HPLC-RIA. In: Touyz, R., Schiffrin, E. (eds) Hypertension. Methods in Molecular Biology, vol 1527. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6625-7_7
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DOI: https://doi.org/10.1007/978-1-4939-6625-7_7
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