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首页> 外文期刊>American Journal of Physiology >Cellular localization of adenine receptors in the rat kidney and their functional significance in the inner medullary collecting duct.
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Cellular localization of adenine receptors in the rat kidney and their functional significance in the inner medullary collecting duct.

机译：大鼠肾中腺嘌呤受体的细胞定位及其在内髓内收集管道中的功能意义。

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The Gi-coupled adenine receptor (AdeR) binds adenine with high affinity and potentially reduces cellular cAMP levels. Since cAMP is an important second messenger in the renal transport of water and solutes, we localized AdeR in the rat kidney. Real-time RT-PCR showed higher relative expression of AdeR mRNA in the cortex and outer medulla compared with the inner medulla. Immunoblots using a peptide-derived and affinity-purified rabbit polyclonal antibody specific for an 18-amino acid COOH-terminal sequence of rat AdeR, which we generated, detected two bands between -30 and 40 kDa (molecular mass of native protein: 37 kDa) in the cortex, outer medulla, and inner medulla. These bands were ablated by preadsorp-tion of the antibody with the immunizing peptide. Immunofluores-cence labeling showed expression of AdeR protein in all regions of the kidney. Immunoperoxidase revealed strong labeling of AdeR protein in the cortical vasculature, including the glomerular arterioles, and less intense labeling in the cells of the collecting duct system. Confocal immunofluorescence imaging colocalized AdeR with aqua-porin-2 protein to the apical plasma membrane in the collecting duct. Functionally, adenine (10 μM) significantly decreased (P < 0.01) 1-deamino-8-D-arginine vasopressin (10 nM)-induced cAMP production in ex vivo preparations of inner medullary collecting ducts, which was reversed by PSB-08162 (20 μM, P < 0.01), a selective antagonist of AdeR. Thus, we demonstrated the expression of AdeR in the renal vasculature and collecting ducts and its functional relevance. This study may open a new avenue for the exploration of autocrine/ paracrine regulation of renal vascular and tubular functions by the nucleobase adenine in health and disease.
机译：Gi偶联的腺嘌呤受体（涂剂）与高亲和力结合腺嘌呤，并且可能降低细胞阵营水平。由于阵营是肾脏运输中的重要第二个使者，我们在大鼠肾脏局部涂抹着剂。与内髓质相比，实时RT-PCR显示皮质和外髓内的涂剂mRNA的相对相对表达。使用肽衍生的和亲和纯化的兔多克隆多克隆多克隆抗体，其针对我们产生的大鼠涂剂的18-氨基酸官序列，检测到-30至40kDa之间的两个带（天然蛋白质：37kDa的分子量）在皮质，外髓质和内髓质。通过用免疫肽的抗体的预溶剂，这些条带被烧蚀。免疫功能标记显示肾脏的所有区域中的涂剂蛋白表达。免疫氧化酶揭示了皮质脉管系统中的涂剂蛋白的强烈标记，包括肾小球动脉瘤，并且在收集管道系统的细胞中较小标记。将共聚焦免疫荧光成像与Aqua-Porin-2蛋白的分层粘合剂分配到收集管中的顶端血浆膜中。在功能上，腺嘌呤（10μm）显着降低（P <0.01）1-达芯-8-D-精氨酸血压加压素（10nm）诱导的内髓内收集管道的exvivo制剂中的营养蛋白生产，由PSB-08162逆转（ 20μm，p <0.01），一种选择性拮抗剂。因此，我们证明了在肾脉管系统和收集管道中的涂剂的表达及其功能相关性。本研究可以通过在健康和疾病中，核心腺嘌呤腺嘌呤开辟探索肾脏血管和管状功能的自分泌/旁静脉调节的新途径。

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