The cellular distribution of Ca(2+)-inhibitable adenylyl cyclase (AC) type 5 and type 6 mRNAs in rat outer medullary collecting duct (OMCD) was performed by in situ hybridization. Kidney sections were also stained with specific antibodies against either collecting duct intercalated cells or principal cells. The localization of type 5 AC in H(+)-ATPase-, but not aquaporin-3-, positive cells demonstrated that type 5 AC mRNA is expressed only in intercalated cells. In contrast, type 6 AC mRNA was observed in both intercalated and principal cells. In microdissected OMCDs, the simultaneous superfusion of carbachol and PGE(2) elicited an additive increase in the intracellular Ca(2+) concentration, suggesting that the Ca(2+)-dependent regulation of these agents occurs in different cell types. Glucagon-dependent cAMP synthesis was inhibited by both a pertussis toxin-sensitive PGE(2) pathway (63.7 +/- 4.6% inhibition, n = 5) and a Ca(2+)-dependent carbachol pathway (48.6 +/- 3.3%, n = 5). The simultaneous addition of both agents induced a cumulative inhibition of glucagon-dependent cAMP synthesis (78.2 +/- 3.3%, n = 5). The results demonstrate a distinct cellular localization of type 5 and type 6 AC mRNAs in OMCD and the functional expression of these Ca(2+)-inhibitable enzymes in intercalated cells.

译文

通过原位杂交进行了大鼠外髓质收集管 (OMCD) 中Ca(2) 抑制腺苷酸环化酶 (AC) 5型和6型mrna的细胞分布。肾脏切片也用针对收集导管插层细胞或主要细胞的特异性抗体染色。5型AC在H(+)-ATPase-而非aquaporin-3-阳性细胞中的定位表明5型AC mRNA仅在插层细胞中表达。相反,在插层细胞和主细胞中均观察到6型AC mRNA。在显微解剖的OMCDs中,卡巴胆碱和PGE(2) 的同时融合引起细胞内Ca(2) 浓度的增加,表明这些试剂的Ca(2) 依赖性调节发生在不同的细胞类型中。胰高血糖素依赖性cAMP合成被百日咳毒素敏感的PGE(2) 途径 (63.7 +/- 4.6% 抑制,n = 5) 和Ca(2 +) 依赖性卡巴胆碱途径 (48.6 +/- 3.3%,n = 5) 抑制。两种药物的同时加入诱导了胰高血糖素依赖性cAMP合成的累积抑制 (78.2 +/- 3.3%,n = 5)。结果表明,OMCD中5型和6型AC mrna具有明显的细胞定位,并且这些Ca(2) 抑制酶在插入细胞中的功能表达。

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