Isolated superfused juxtaglomerular cells from rat kidney: a model for study of renin secretion

Abstract

Freshly isolated rat juxtaglomerular cells (JGC) were superfused to study renin secretion rate (RSR) at the cellular level. Effluates from the superfusion chamber collected in 20-min intervals showed a time-dependent decline in RSR from 85.5 ± 32 to 4.0 ± 2.4 ng ANG I ⋅ ml−1 ⋅ h−1 ⋅ mg protein−1 ⋅ min−1within 100 min of collection (mean ± SE, n = no. of JGC preparations/superfusion chambers = 9/18). Addition of adenosine deaminase type II (ADA II, 3 U/1.4 mg protein) to the superfusion medium increased RSR more than fourfold to 402 ± 100 ng in the first collection period, which dropped to 237.5 ± 67 ng ANG I ⋅ ml−1 ⋅ h−1 ⋅ mg protein−1 ⋅ min−1( n = 9/18) within 100 min. This ADA II effect was rapid in onset and fully reversible. When the purified ADA type VII, with a 40-fold higher specific activity, was added to the superfusate, RSR was increased only by 96 ± 17.8% compared with controls. This ADA VII (5 U/30 μg) effect could be mimicked by the selective adenosine A1-receptor antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX, 10−6 mol/l). Since albumin stimulated RSR in a concentration-dependent fashion, to an extent similar to that of ADA II, we assume that the ADA II effect was largely unspecific in nature. We conclude that 1) superfusion of isolated JGC from rats is suitable for investigations of renin secretion at the cellular level, 2) the increase in RSR by ADA II appears to be only in part due to deamination of endogenously generated adenosine, and 3) albumin in the superfusate induces a similar stimulatory effect as ADA II.