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Renal renin secretion is regulated by numerous physiological factors which act in a direct or indirect fashion through alterations in hemodynamic or circulatory factors. Renin release studies using renal cortical slices have been reported in the rat, rabbit, dog, pig, and recently in the mouse of examine the direct stimuli on juxtaglomerullar cells not mediated by systemic effects. However, the evaluations of some of the essential characters of the experimental system, e.g., energy dependency (1, 2), Ca dependency (2-4), and oxygen demand (5, 6) of renin secretion show discrepancies in some of the studies. Me thodological problems seems to be involved in some of the inconsistencies in the experi mental results. Renin determination in many studies have been performed through angiotensin I radioimmunoassay since Haber et al. (7). Commercially available angiotensin I radioimmunoassay kits are, however, expensive and sometimes insufficient in the sensitivity for various pharmacological studies. In the present paper, we will describe the preparation of anti-angiotensin I anti serum, the determination of renin activity released from dog renal cortical slices by radioimmunoassay of angiotensin I, and adrenergic 3-receptor mediated renin release using this experimental system. For the primary and secondary immuni zation, angiotensin I antigen was synthesized by mixing 10 mg each of angiotensin I (Sigma, A9650: Isoleucine5 angiotensin I) and bovine serum albumin in 0.5 ml of water and then adding of 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide HCI (EDC) (150 mg) in 0.5 ml of water. After 30 min standing at room temperature, the remarkably ag gragated reaction mixture was dialyzed against distilled water, lyophilized and kept until use. Five mg of the conjugate was dispersed by sonication in saline (1 ml), emulsified with an equal volume of Freund's complete adjuvant, and injected into two male rabbits in two thigh sites (i.m.) and three sites on the back (s.c.). Three booster injections were performed three, eight, and seventeen months after the primary injection. In the antigen synthesis for the third and fourth injections, the amount of EDC was reduced to 10 mg and 50 mg, respectively. Selection of antiserum for a sensitive radioimmunoassay was performed by examining the 1251 angiotensin I binding capacity curves of the antisera at a dilution-fold of 50% binding. As shown in Table 1, a dissociation between high titer and high sensitivity was observed. All antisera obtained after the third immuni zation were available for an angiotensin I radioimmunoassay in this study. Specificity of these …

Stimulation of renin release from dog renal cortical slices with L-isoproterenol and dibutyryl cyclic adenosine 3',5'-monophosphate.