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Am J Physiol Renal Physiol 294: F1257-F1263, 2008. First published March 19, 2008; doi:10.1152/ajprenal.00588.2007
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INNOVATIVE METHODOLOGY

Determination of plasma and urinary angiotensinogen levels in rodents by newly developed ELISA

Hiroyuki Kobori,1 Akemi Katsurada,1 Kayoko Miyata,1 Naro Ohashi,1 Ryousuke Satou,1 Toshie Saito,1 Yoshiaki Hagiwara,2 Kazuya Miyashita,2 and L. Gabriel Navar1

1Department of Physiology, and Hypertension and Renal Center of Excellence, Tulane University Health Sciences Center, New Orleans, Louisiana; and 2Immuno-Biological Laboratories, Fujioka, Gunma, Japan

Submitted 11 December 2007 ; accepted in final form 17 March 2008

We recently reported that urinary excretion rates of angiotensinogen provide a specific index of the intrarenal renin-angiotensin system status in angiotensin II-dependent hypertensive rats. Angiotensinogen concentrations in mouse plasma are thought to be much lower than those in rat plasma; however, detailed information is deficient due to lack of direct quantitative measurements of rodent angiotensinogen. To elucidate this issue, we have developed a quantitative method for measurement of rodent angiotensinogen using a sandwich-type ELISA. The standard curve for mouse and rat angiotensinogen exhibited a high linearity at 0.16–10 and 0.08–5 ng/ml, respectively, with correlation coefficients >0.99. While plasma angiotensinogen concentrations of male high serum IgA (HIGA) mice (IgA nephritis model animals, 1,308 ± 47 ng/ml; n = 10) were lower than those of control BALB/c mice (1,620 ± 384; n = 12), urinary angiotensinogen concentrations of HIGA mice (14.6 ± 1.5 ng/ml; n = 34) were higher than those of BALB/c mice (4.6 ± 0.1; n = 2). In a similar manner, while plasma angiotensinogen concentrations of Zucker diabetic fatty (ZDF) obese rats (type 2 diabetic model animals, 1,789 ± 50 ng/ml; n = 5) were lower than those of control ZDF lean rats (2,296 ± 47; n = 5), urinary angiotensinogen concentrations of ZDF obese rats (88.2 ± 11.4 ng/ml; n = 15) were higher than those of ZDF lean rats (31.3 ± 1.9; n = 15). These data indicate that plasma and urinary angiotensinogen concentrations are less in mice than rats. However, these data suggest that urinary angiotensinogen levels are different from plasma angiotensinogen levels in rodents. The development of rodent angiotensinogen ELISA allows quantitative comparisons in mouse and rat angiotensinogen levels in models of hypertension and cardiovascular and kidney diseases.

enzyme-linked immunosorbent assay; renin-angiotensin system; plasma; urine; mouse; rat


Address for reprint requests and other correspondence: H. Kobori, Depts. of Medicine and Physiology and Hypertension and Renal Center of Excellence, Tulane Univ. Health Sciences Center, 1430 Tulane Ave., #SL39, New Orleans, LA 70112-2699 (e-mail: hkobori{at}tulane.edu)






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