Vasopressin and cAMP stimulate electrogenic chloride secretion in an IMCD cell line.
Vasopressin and cAMP stimulate electrogenic chloride secretion in an IMCD cell line.
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加压素和 cAMP 刺激 IMCD 细胞系中的电化学氯化物分泌。
DOI：
10.1152/ajprenal.1995.268.5.f854
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Stanton,BA
中科院分区：
文献类型：
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作者：
Kizer,NL;Vandorpe,D;Lewis,B;Bunting,B;Russell,J;Stanton,BA
此前，我们证明源自转基因小鼠内髓集合管（IMCD）的 mIMCD-K2 细胞系通过生电机制分泌 Cl- [N. L. Kizer、B. Lewis 和 B. A. Stanton, Am。 J.生理学。 268（肾液电解质生理学。37）：F347-F355，1995]。本研究的目的是表征电生 Cl 分泌 (IscCl) 的细胞机制，并确定精氨酸加压素 (AVP) 和腺苷 3',5'-环单磷酸 (cAMP) 是否刺激 IscCl。为此，我们测量了安装在 Ussing 型室中的 mIMCD-K2 细胞单层的 IscCl。 AVP 增加 IscCl，米氏常数 (Km) 为 2.1 +/- 0.7 x 10(-12) M。1-Desamino-8-D-AVP（一种特定的 V2 受体激动剂）可将 IscCl 从 3.3 +/- 0.4 增加到 17.4 +/- 1.3 microA/cm2，8-(4-氯苯硫基)-cAMP（一种细胞永久类似物） cAMP（AVP 的第二信使）的 cAMP 使 IscCl 从 1.4 +/- 0.3 增加到 15.2 +/- 1.2 microA/cm2。呋塞米和布美他尼（Na(+)-2Cl(-)-K+ 共转运抑制剂）和 4,4'-二异硫氰芪-2,2'-二磺酸 (DIDS)（Cl-/HCO3- 交换抑制剂）添加到基底外侧溶液中时，可减少 IscCl。我们的数据表明，AVP 通过 V2 受体和第二信使 cAMP 刺激 IscCl，mIMCD-K2 细胞的 Cl- 分泌涉及通过 Na(+)-2Cl(-)-K+ 共转运跨基底外侧膜摄取 Cl-，以及通过囊性纤维化跨膜电导调节器 Cl- 通道跨顶膜交换和扩散出细胞。
Previously, we demonstrated that the mIMCD-K2 cell line, derived from the inner medullary collecting duct (IMCD) of a transgenic mouse, secretes Cl- by an electrogenic mechanism [N. L. Kizer, B. Lewis, and B. A. Stanton, Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F347-F355, 1995]. The objective of the present study was to characterize the cellular mechanisms of electrogenic Cl- secretion (IscCl) and to determine whether arginine vasopressin (AVP) and adenosine 3',5'-cyclic monophosphate (cAMP) stimulate IscCl. To this end, we measured IscCl across monolayers of mIMCD-K2 cells mounted in Ussing-type chambers. AVP increased IscCl with a Michaelis constant (Km) of 2.1 +/- 0.7 x 10(-12) M. 1-Desamino-8-D-AVP, a specific V2 receptor agonist, increased IscCl from 3.3 +/- 0.4 to 17.4 +/- 1.3 microA/cm2, 8-(4-Chlorophenylthio)-cAMP, a cell-permanent analogue of cAMP, a second messenger of AVP, increased IscCl from 1.4 +/- 0.3 to 15.2 +/- 1.2 microA/cm2. Furosemide and bumetanide, inhibitors of Na(+)-2Cl(-)-K+ cotransport, and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), an inhibitor of Cl-/HCO3- exchange, reduced IscCl when added to the basolateral solution. Our data suggest that AVP, via V2 receptors, and the second messenger cAMP stimulate IscCl and that Cl- secretion by mIMCD-K2 cells involves uptake of Cl- across the basolateral membrane by Na(+)-2Cl(-)-K+ cotransport and Cl-/HCO3- exchange and diffusion out of the cells across the apical membrane by cystic fibrosis transmembrane conductance regulator Cl- channels.