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首页> 外文期刊>American Journal of Physiology >Capacitative calcium entry and TRPC channel proteins are expressed in rat distal pulmonary arterial smooth muscle.
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Capacitative calcium entry and TRPC channel proteins are expressed in rat distal pulmonary arterial smooth muscle.

机译：电容性钙进入和TRPC通道蛋白在大鼠远端肺动脉平滑肌中表达。

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Mammalian homologs of transient receptor potential (TRP) genes in Drosophila encode TRPC proteins, which make up cation channels that play several putative roles, including Ca2+ entry triggered by depletion of Ca2+ stores in endoplasmic reticulum (ER). This capacitative calcium entry (CCE) is thought to replenish Ca2+ stores and contribute to signaling in many tissues, including smooth muscle cells from main pulmonary artery (PASMCs); however, the roles of CCE and TRPC proteins in PASMCs from distal pulmonary arteries, which are thought to be the major site of pulmonary vasoreactivity, remain uncertain. As an initial test of the possibility that TRPC channels contribute to CCE and Ca2+ signaling in distal PASMCs, we measured [Ca2+]i by fura-2 fluorescence in primary cultures of myocytes isolated from rat intrapulmonary arteries (>4th generation). In cells perfused with Ca2+-free media containing cyclopiazonic acid (10 microM) and nifedipine (5 microM) to deplete ER Ca2+ stores and block voltage-dependent Ca2+ channels, restoration of extracellular Ca2+ (2.5 mM) caused marked increases in [Ca2+]i whereas MnCl2 (200 microM) quenched fura-2 fluorescence, indicating CCE. SKF-96365, LaCl3, and NiCl2, blocked CCE at concentrations that did not alter Ca2+ responses to 60 mM KCl (IC50 6.3, 40.4, and 191 microM, respectively). RT-PCR and Western blotting performed on RNA and protein isolated from distal intrapulmonary arteries and PASMCs revealed mRNA and protein expression for TRPC1, -4, and -6, but not TRPC2, -3, -5, or -7. Our results suggest that CCE through TRPC-encoded Ca2+ channels could contribute to Ca2+ signaling in myocytes from distal intrapulmonary arteries.
机译：果蝇中的瞬时受体电位（TRP）基因的哺乳动物同源物编码TRPC蛋白，该蛋白构成起多种假定作用的阳离子通道，包括由内质网（ER）中的Ca2 +储存耗尽触发的Ca2 +进入。这种电容性钙离子进入（CCE）被认为可以补充Ca2 +的储存，并有助于许多组织的信号传导，包括来自主要肺动脉（PASMC）的平滑肌细胞。然而，CCE和TRPC蛋白在远端肺动脉PASMC中的作用仍不确定，据认为这是肺血管反应性的主要部位。作为TRPC通道在远端PASMCs中促进CCE和Ca2 +信号传导的可能性的初步测试，我们通过fura-2荧光在从大鼠肺动脉（>第4代）分离的心肌细胞的原代培养物中测量了[Ca2 +] i。在用不含Ca2 +的培养基灌注的细胞中，该培养基不含环吡唑酸（10 microM）和硝苯地平（5 microM）来耗尽ER Ca2 +的储存并阻断电压依赖性Ca2 +通道，细胞外Ca2 +（2.5 mM）的恢复导致[Ca2 +] i显着增加。而MnCl2（200 microM）淬灭了fura-2荧光，表明CCE。 SKF-96365，LaCl3和NiCl2在不会改变Ca2 +对60 mM KCl的浓度（分别为IC50 6.3、40.4和191 microM）下阻断CCE。对从远端肺内动脉和PASMCs分离的RNA和蛋白进行的RT-PCR和Western blotting显示，TRPC1，-4和-6的mRNA和蛋白表达，但TRPC2，-3，-5或-7则不。我们的研究结果表明，CCE通过TRPC编码的Ca2 +通道可能有助于远端肺动脉内肌细胞的Ca2 +信号传导。

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