Hill Andrew J, Hinton Jane M, Cheng Hongwei, Gao Zhan, Bates David O, Hancox Jules C, Langton Philip D, James Andrew F
Cardiovascular and Microvascular Research Laboratories, Department of Physiology, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, UK.
Cell Calcium. 2006 Jul;40(1):29-40. doi: 10.1016/j.ceca.2006.03.007. Epub 2006 May 11.
The TRPC family of non-selective cation channels has been suggested to play a key role in the responses to alpha1-adrenoceptor stimulation of vascular smooth muscle. However, there are still very few reports of non-selective cation currents activated by alpha1-AR in resistance arteries. Here, we examine the expression of TRPC channels and the currents activated by alpha1-adrenoceptors in rat mesenteric resistance artery smooth muscle. Messenger RNA and protein for TRPC1, TRPC3 and TRPC6 were detected within the arteries by RT-PCR and immunoblotting. Endothelial and adventitial layers were found to express the TRPC1, TRPC3 and TRPC6 proteins whereas only TRPC1 and TRPC6 were detected in the arterial smooth muscle by confocal immunofluorescence microscopy. In whole-cell patch-clamp recordings from isolated mesenteric arterial myocytes, an outwardly rectifying non-selective cation current was activated by both the alpha1-adrenoceptor agonist, phenylephrine (10 microM), and the diacylglycerol analogue, 1-oleoyl-2-acetyl-sn-glycerol (100 microM). Responses to 1-oleoyl-2-acetyl-sn-glycerol were not blocked, but increased, following inhibition of protein-kinase-C with either bisindolylmaleimide-I (1 microM) or chelerythrine (1 microM). The currents activated by both phenylephrine and 1-oleoyl-2-acetyl-sn-glycerol were inhibited by Gd3+ (100 microM) but potentiated by flufenamic acid (100 microM). Collectively, these findings demonstrate for the first time the expression of TRPC1 and TRPC6 in rat mesenteric artery smooth muscle and the existence in rat isolated mesenteric arterial myocytes of a TRPC-like non-selective cation current activated by alpha1-adrenoceptor stimulation and 1-oleoyl-2-acetyl-sn-glycerol.
非选择性阳离子通道的瞬时受体电位经典型(TRPC)家族被认为在血管平滑肌对α1 -肾上腺素能受体刺激的反应中起关键作用。然而,关于α1 -肾上腺素能受体在阻力动脉中激活非选择性阳离子电流的报道仍然非常少。在此,我们研究了TRPC通道在大鼠肠系膜阻力动脉平滑肌中的表达以及α1 -肾上腺素能受体激活的电流。通过逆转录聚合酶链反应(RT-PCR)和免疫印迹法在动脉内检测到了TRPC1、TRPC3和TRPC6的信使核糖核酸(mRNA)和蛋白质。发现内皮和外膜层表达TRPC1、TRPC3和TRPC6蛋白,而通过共聚焦免疫荧光显微镜在动脉平滑肌中仅检测到TRPC1和TRPC6。在分离的肠系膜动脉肌细胞的全细胞膜片钳记录中,α1 -肾上腺素能受体激动剂去氧肾上腺素(10微摩尔)和二酰基甘油类似物1-油酰基-2-乙酰基-sn-甘油(100微摩尔)均可激活外向整流的非选择性阳离子电流。在用双吲哚马来酰亚胺-I(1微摩尔)或白屈菜红碱(1微摩尔)抑制蛋白激酶C后,对1-油酰基-2-乙酰基-sn-甘油的反应未被阻断,反而增强。去氧肾上腺素和1-油酰基-2-乙酰基-sn-甘油激活的电流均被钆离子(100微摩尔)抑制,但被氟芬那酸(100微摩尔)增强。总的来说,这些发现首次证明了TRPC1和TRPC6在大鼠肠系膜动脉平滑肌中的表达,以及在大鼠分离的肠系膜动脉肌细胞中存在一种由α1 -肾上腺素能受体刺激和1-油酰基-2-乙酰基-sn-甘油激活的TRPC样非选择性阳离子电流。
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