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首页> 美国卫生研究院文献>Biochemical Journal >Cholera-toxin and corticotropin modulation of inositol phosphate accumulation induced by vasopressin and angiotensin II in rat glomerulosa cells.
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Cholera-toxin and corticotropin modulation of inositol phosphate accumulation induced by vasopressin and angiotensin II in rat glomerulosa cells.

机译：血管加压素和血管紧张素II在大鼠肾小球细胞中诱导的霍乱毒素和促肾上腺皮质激素对肌醇磷酸积累的调节。

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Vasopressin (VP) and angiotensin II (AT II) stimulate the production of inositol phosphates (IP) in rat glomerulosa cells. Guanosine 5'-[gamma-thio]triphosphate (GTP[S]), but not VP or AT II, stimulates IP production in a myo-[3H]inositol-prelabelled glomerulosa-cell membrane preparation. In combination with GTP[S], these hormones potentiate the response to GTP[S], indicating the existence of a G-protein involved in the coupling of the VP and AT II receptor with the phospholipase C. ADP-ribosylation with pertussis toxin (IAP) revealed the specific labelling of a single molecule of 41 kDa. No significant inhibition of VP- or AT II-stimulated IP accumulation was detected in intact cells when the whole 41 kDa molecule was endogenously ADP-ribosylated by IAP treatment. On the contrary, when glomerulosa cells were infected with cholera toxin (CT), both the VP- and AT II-stimulated IP accumulations were inhibited in a dose-dependent manner. Yet these effects were partial even at high concentrations of CT, and could not be related to the ADP-ribosylation of 'alpha s' molecules. Similarly, when the cells were infected with 1 microgram of CT/ml, the specific binding of VP and AT II decreased by 50-60%. Such results may signify that the treatment primarily affects the densities of the hormone receptors. When glomerulosa cells were incubated for 15 h in the presence of 10 nM-corticotropin (ACTH), a condition in which the intracellular concentration of cyclic AMP was increased 3-fold, the maximum IP response to 0.1 microM-VP or -AT II was decreased by 50%. When similar experiments were carried out only after a 15 min incubation period with the same concentration of ACTH, the increase in cyclic AMP was more pronounced, but no inhibition of hormone-induced IP accumulation was observed. Altogether, these results may suggest that CT exerts its action on the VP- or AT II-sensitive phospholipase C systems via a prolonged increase in intracellular cyclic AMP.
机译：加压素（VP）和血管紧张素II（AT II）刺激大鼠肾小球细胞中肌醇磷酸酯（IP）的产生。鸟苷5'-[γ-硫代]三磷酸酯（GTP [S]），而不是VP或AT II，可刺激肌-[3H]肌醇预标记的肾小球细胞膜制剂中IP的产生。与GTP [S]结合使用时，这些激素增强了对GTP [S]的反应，表明存在VP和AT II受体与磷脂酶C偶联的G蛋白。ADP-核糖基化与百日咳毒素（ IAP）揭示了41 kDa单分子的特异性标记。当通过IAP处理将整个41 kDa分子内源性ADP核糖基化时，在完整细胞中未检测到对VP或AT II刺激的IP积累的显着抑制。相反，当肾小球细胞感染霍乱毒素（CT）时，VP和AT II刺激的IP积累均受到剂量依赖性的抑制。然而，即使在高浓度的CT下，这些作用仍是部分的，并且与“αs”分子的ADP-核糖基化无关。同样，当细胞被1微克CT / ml感染时，VP和AT II的特异性结合降低了50-60％。这样的结果可能表明该治疗主要影响激素受体的密度。当肾小球细胞在10 nM促肾上腺皮质激素（ACTH）存在下孵育15小时时，细胞内环状AMP浓度增加3倍的条件下，对0.1 microM-VP或-AT II的最大IP反应为减少了50％。当仅在相同浓度的ACTH孵育15分钟后进行类似实验时，环AMP的增加更为明显，但未观察到激素诱导的IP积累受到抑制。总而言之，这些结果可能表明CT通过延长细胞内环AMP的作用在VP或AT II敏感的磷脂酶C系统上发挥作用。

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