Apical-basolateral membrane asymmetry in canine cortical collecting tubule cells. Bradykinin, arginine vasopressin, prostaglandin E2 interrelationships.
Apical-basolateral membrane asymmetry in canine cortical collecting tubule cells. Bradykinin, arginine vasopressin, prostaglandin E2 interrelationships.
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犬皮质集合管细胞的顶基底外侧膜不对称。
DOI：
10.1172/jci111419
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
Smith,WL
中科院分区：
文献类型：
--
作者：
Garcia-Perez,A;Smith,WL
本文报道的研究旨在确定前列腺素在肾集合小管的释放或前列腺素在肾集合小管中的生化作用是否存在根尖-基底侧不对称。犬皮质收集小管（CCCT）细胞通过免疫解剖分离，并以超汇合密度在Millipore过滤器上播种。形成的CCCT细胞融合单层：(a)形成并维持了1 mV（顶侧负）的跨细胞电位差；(b)对菊粉的渗透性与Madin-Darby犬肾（MDCK）细胞类似单层的渗透性相同；(c)当精氨酸加压素（AVP）加入到细胞膜基底外侧而非顶端表面时，释放腺苷3′，5′-环单磷酸腺苷（cAMP）。这些结果表明，微孔滤膜上的CCCT细胞的汇合单层具有不对称的特征，与完整的收集小管一样。此外，添加到CCCT细胞单层两侧的PGE2以与菊粉相同的慢速穿过单层，证明了检测PGE2作用的侧边性和释放的可行性。虽然AVP仅在CCCT细胞基底外侧释放cAMP，但AVP在根尖或基底外侧表面均可引起PGE2的释放。这一结果表明至少存在两种AVP受体系统，一种与cAMP合成偶联，另一种与PGE2形成偶联。与AVP观察到的结果相反，缓激肽仅当添加到CCCT细胞的顶端表面时才引起PGE2的释放，这表明尿源性而非血源性的激肽通过犬收集小管诱导PGE2的形成。在AVP和缓激肽作用下，PGE2在单分子膜两侧的释放量相当。高浓度（大于或等于10(-8)M）的PGE2加入到单分子膜的两侧，引起cAMP的释放。然而，在浓度(10（-10)-10 (-12)M）时，PGE2对cAMP的释放没有独立影响，PGE2抑制了cAMP的释放，这通常是对AVP的反应。这种抑制发生在PGE2添加到CCCT细胞单层的顶端或底外侧表面时。PGE2 （10(-11) M）也能抑制avp诱导的CCCT细胞细胞内cAMP的积累。这种抑制作用仅在细胞与PGE2预孵育大于或等于20分钟时观察到。我们的结果与前列腺素抑制AVP的水渗透作用是由于抑制AVP诱导的cAMP产生的概念是一致的。这种抑制似乎不涉及PGE2与AVP受体（与腺苷酸环化酶偶联）的直接物理相互作用，因为CCCT细胞必须与PGE2预孵育20分钟才能观察到抑制作用，并且由于添加到CCCT细胞的顶表面的PGE2抑制cAMP释放，以响应从基底外侧表面作用的AVP。图片
The studies reported here were designed to determine if there is an apical-basolateral asymmetry to the release of prostaglandins by or to the biochemical effects of prostaglandins on the renal collecting tubule. Canine cortical collecting tubule (CCCT) cells were isolated by immunodissection and seeded at supraconfluent densities on Millipore filters. The resulting confluent monolayer of CCCT cells: (a) developed and maintained a transcellular potential difference of 1 mV (apical side negative); (b) exhibited a permeability to inulin that was the same as that obtained with similar monolayers of Madin-Darby canine kidney (MDCK) cells; and (c) released adenosine 3',5'-cyclicmonophosphate (cAMP) in response to arginine vasopressin (AVP) added to the basolateral but not the apical surface of the monolayer. These results indicate that confluent monolayers of CCCT cells on Millipore filters have characteristics of asymmetry that are seen with intact collecting tubules. Moreover, PGE2 added to either side of the CCCT cell monolayer crossed the monolayer at the same slow rate as inulin, which demonstrated the feasibility of examining the sidedness of the effects of and the release of PGE2. Although AVP caused cAMP release only when added to the basolateral side of CCCT cells, AVP caused the release of PGE2 when added to either the apical or basolateral surface. This result implies that there are at least two AVP receptor systems, one coupled to cAMP synthesis and one to PGE2 formation. In contrast to the results observed with AVP, bradykinin caused PGE2 release only when added to the apical surface of CCCT cells, which suggested that urinary but not blood borne kinins elicit PGE2 formation by the canine collecting tubule. PGE2 was released in comparable amounts on each side of the monolayer in response both to AVP and to bradykinin. High concentrations (greater than or equal to 10(-8) M) of PGE2 added to either side of the monolayer caused the release of cAMP. However, at concentrations (10(-10) - 10(-12) M) at which PGE2 had no independent effect on cAMP release, PGE2 inhibited the release of cAMP, which normally occurred in response to AVP. This inhibition occurred with PGE2 added to either the apical or basolateral surface of the CCCT cell monolayer. PGE2 (10(-11) M) also inhibited the AVP-induced accumulation of intracellular cAMP by CCCT cells seeded on culture dishes. This inhibition was only observed when the cells were preincubated with PGE2 for greater than or equal to 20 min. Our results are consistent with the concept that inhibiton by prostaglandins of the hydroosmotic effect of AVP is due to inhibition of AVP-induced cAMP production. This inhibition does not appear to involve a direct physical interaction of PGE2 with the AVP receptor which is coupled to adenylate cyclase, since CCCT cells must be preincubated with PGE2 for 20 min for the inhibition to be observed, and since PGE2 added to the apical surface of CCCT cells inhibits cAMP release in response to AVP acting from the basolateral surface.Images