Rightsel W A, Okamura T, Inagami T, Pitcock J A, Takii Y, Brooks B, Brown P, Muirhead E E
Circ Res. 1982 Jun;50(6):822-9. doi: 10.1161/01.res.50.6.822.
A monolayer cell culture of juxtaglomerular cells (JGC) was derived from the renal cortex of neonatal rats. The JGC had the characteristics of those within the kidney, including peripheral dense bodies and myofibrils indicating a smooth muscle origin; rough ER containing fluffy material consistent with protein synthesis; a prominent Golgi apparatus for packaging granules, and granules having the characteristics of secretory granules and lysosomes. Transplants of the cultured cells into syngeneic recipients survived for 2 weeks or longer and retained the features of JGC. The JGC granules fluoresced when treated with a rabbit antibody against pure rat renin, followed by fluorescein isothyocyanate conjugated F(ab')2 fragment of goat antirabbit IgG (Fc fragment) heavy chain specific. The latter indicated the presence of renin. The JGC were lysed in the presence of DFP, captopril, leupeptin, and EDTA, and were extracted in the presence of pepstatin. The lysate contained renin activity that was inhibited by a specific renin antibody. Nonspecific proteases were excluded by the antibody and its pH optimum. Angiotensin I-converting enzyme was detected in the lysate prepared without the use of EDTA and captopril. Angiotensins I and II/III were derived from the extract by additional extractions, TLC, and RIA, using highly specific antibodies. The angiotensins were confirmed by chromatography monitored by authentic angiotensins. We concluded that the cultured JGC contained renin, angiotensin I-converting enzyme, and angiotensin I and II/III.
近球细胞(JGC)单层细胞培养物源自新生大鼠的肾皮质。这些近球细胞具有肾内近球细胞的特征,包括外周致密体和肌原纤维,表明其起源于平滑肌;含有与蛋白质合成一致的蓬松物质的粗面内质网;用于包装颗粒的显著高尔基体,以及具有分泌颗粒和溶酶体特征的颗粒。将培养的细胞移植到同基因受体中可存活2周或更长时间,并保留近球细胞的特征。用抗纯大鼠肾素的兔抗体处理近球细胞颗粒后发荧光,随后用异硫氰酸荧光素偶联的山羊抗兔IgG(Fc片段)重链特异性F(ab')2片段处理。后者表明存在肾素。近球细胞在二异丙基氟磷酸(DFP)、卡托普利、亮抑酶肽和乙二胺四乙酸(EDTA)存在的情况下被裂解,并在胃蛋白酶抑制剂存在的情况下被提取。裂解物中含有被特异性肾素抗体抑制的肾素活性。抗体及其最适pH排除了非特异性蛋白酶。在不使用EDTA和卡托普利制备的裂解物中检测到血管紧张素I转换酶。通过额外的提取、薄层层析(TLC)和放射免疫分析(RIA),使用高度特异性抗体从提取物中获得血管紧张素I和II/III。通过用标准血管紧张素监测的层析法证实了血管紧张素。我们得出结论,培养的近球细胞含有肾素、血管紧张素I转换酶以及血管紧张素I和II/III。