A Radioimmunoassay for the Direct Measurement of Renin in Mice and Its Application to Submaxillary Gland and Kidney Studies

Michelakis, Andrew M.; Yoshida, Hisanobu; Menzie, James W.; Murakami, Kazuo; Inagami, Tadashi

doi:10.1210/endo-94-4-1101

Cited by 91 publications

(27 citation statements)

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“…Previous investigators (Deodhar et al, 1964;Haas et al, 1962;Haas et al, 1965;Haas et aL, 1975;Helmer, 1958;Lucas et al, 1975;Poulsen, 1977a, 1977b;Michelakis et al, 1974;Peart et al, 1966;Skeggs et al, 1975;Wakerlin et al, 1953;Walsh et at., 1976: Weiser et al, 1969 have used renin preparations of variable degrees of purity as an antigen without further modification except for emulsification in Freund's adjuvant. It is known that chemical alteration of poorly immunogenic molecules, which usually are of relatively low molecular weight, can markedly enhance their antigenicity.…”

Section: Discussionmentioning

confidence: 97%

“…In contrast to the numerous methods available to measure angiotensins I and II, a direct assay for the enzyme renin has been developed only for the mouse submaxillary gland enzyme (Michelakis et al, 1974;Poulsen, 1977a, 1977b). An attempt to establish a direct assay for human renal renin (Lucas et al, 1975) was unsuccessful due to the inhibition of the renin-antibody reaction by a component of plasma.…”

Section: Discussionmentioning

confidence: 99%

“…Due to the impurity of the antigens, these antibodies possessed only limited specificity, which restricted their usefulness. When a homogeneous renin preparation from the mouse submaxillary gland became available (Cohen et al, 1972), antibodies specific for renin were prepared and used to develop a direct radioimmunoasay for the enzyme, which was detected in various tissues of the mouse Poulsen, 1977a, 1977b;Michelakis et al, 1974). However, since antibodies against the mouse submaxillary gland enzyme did not cross-react with renin from other species, this radioimmunoassay was limited to studies in the mouse.…”

mentioning

confidence: 99%

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Specific antibody to hog renal renin and its application to the direct radioimmunoassay of renin in various organs.

Hirose¹,

Workman²,

Inagami³

1979

Circulation Research

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SUMMARY We produced anti-hog renin antibodies using as antigens pure hog renal renin that either had been insolubilized or conjugated to tetanus toxoid. High titer antibodies were obtained, which demonstrated different cross-reactivity with renins from other species. A direct radioimmunoassay for renin was developed using antibody, monoiodinated lu I-hog renin, and various methods for separating free and antibody-bound trace. This assay was capable of detecting 40 pg of hog renin and was applied to the determination of renin in hog blood and other organs. Based on the direct measurement of renin by this radioimmunoassay, the renin-like activity (i.e., the ability to generate angiotensin I from renin substrate preparations) of the pituitary gland was found to be due mostly to true renin, whereas the renin activity of other hog tissues, including the adrenal gland, liver, lung, spleen, and submaxillary gland, was not identified as renin and may have been due to cathepsins. Cire Ret 45: 275-281, 1979

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Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Specific antibody to hog renal renin and its application to the direct radioimmunoassay of renin in various organs.

Hirose¹,

Workman²,

Inagami³

1979

Circulation Research

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“…Secretion of renin by the kidney increases in these conditions and renin in plasma may act upon its substrate to generate AI. However, renin is present in many organs (3)(4)(5)(6) and cell types (7)(8)(9), including vascular smooth muscle cells (8); AI and All and converting enzyme exist in renin-containing cells (7,9,10); and the plasma concentration of All is in the picomolar range (1 1), but binding affinities of this peptide to its receptors are in the nanomolar range (12)(13)(14)(15). These observations raise questions about the role of circulating renin and angiotensin.…”

Section: Introductionmentioning

confidence: 99%

Local generation of angiotensin II as a mechanism of regulation of peripheral vascular tone in the rat.

Oliver¹,

Sciacca²

1984

J. Clin. Invest.

114

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Abstract. Renin is present in vascular smooth muscle cells and has been shown to coexist with angiotensins I (Al) and II (All) in many cell types. Accordingly, we postulated that the renin-angiotensin system controls vascular tone, not by the action of circulating renal renin but rather, by the local generation of angiotensin by vascular renin. Isolated rat hindquarters were perfused in vitro with Krebs-Henseleit buffer containing 7% albumin, and flow-adjusted to obtain a perfusion pressure of -90 mmHg. Infusion of 4.8 nmol* min-' for 5 min of All or Al markedly increased perfusion pressure. An identical dose of the synthetic tetradecaptide of renin substrate (TDCP-RS) increased pressure similarly to Al. The pressure increase evoked by TDCP-RS was markedly decreased by captopril and by two different peptides that inhibit renin. Renin activity in the perfusate, incubated with semipurified rat renin substrate, was 21±3 pg Al ml-' h-' (mean±SEM) at 15 min of perfusion and 47±4 pg Al -ml-' -h-' at 45 min (n = 9; P < 0.01). When TDCP-RS was infused at 4.8 nmol * min-' for 5 min in the presence of captopril, Al in the perfusate increased linearly at a rate of 16.5 pmol min-' for 10 min (n = 5). The results indicate that TDCP-RS constricted the vasculature by its conversion to All and suggest that All was generated from a two-step hydrolysis of TDCP-RS by renin and converting enzyme. The data thus suggest that the renin-angiotensin system controls vascular tone by the local generation of All by renin and converting enzyme in the vasculature.

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“…22 Pure renin and antibodies permitted us to construct a direct radioimmunoassay of renin for the first time. 23 By this method we learned that submaxillary gland renin is not secreted into the plasma in large measure under normal conditions in spite of its enormously high concentration in male mouse submandibular gland (renin content in the salivary gland was approximately 100-fold as high as in the kidney).…”

mentioning

confidence: 99%

Purification of renin and prorenin.

Inagami

1991

Hypertension

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A Radioimmunoassay for the Direct Measurement of Renin in Mice and Its Application to Submaxillary Gland and Kidney Studies

Cited by 91 publications

References 0 publications

Specific antibody to hog renal renin and its application to the direct radioimmunoassay of renin in various organs.

Specific antibody to hog renal renin and its application to the direct radioimmunoassay of renin in various organs.

Local generation of angiotensin II as a mechanism of regulation of peripheral vascular tone in the rat.

Purification of renin and prorenin.

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