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电压依赖性钙通道Cav1.2、2.1a和2.1b在血管肌细胞中的共表达。

Coexpression of voltage-dependent calcium channels Cav1.2, 2.1a, and 2.1b in vascular myocytes.

作者信息

Andreasen Ditte, Friis Ulla G, Uhrenholt Torben R, Jensen Boye L, Skøtt Ole, Hansen Pernille B

机构信息

Physiology and Pharmacology, University of Southern Denmark, Odense, Denmark.

出版信息

Hypertension. 2006 Apr;47(4):735-41. doi: 10.1161/01.HYP.0000203160.80972.47. Epub 2006 Feb 27.

Abstract

Voltage-dependent Ca2+ channels Cav1.2 (L type) and Cav2.1 (P/Q type) are expressed in vascular smooth muscle cells (VSMCs) and are important for the contraction of renal resistance vessels. In the present study we examined whether native renal VSMCs coexpress L-, P-, and Q-type Ca2+ currents. The expression of both Cav2.1a (P-type) and Cav2.1b (Q-type) mRNA was demonstrated by RT-PCR in renal preglomerular vessels from rats and mice. Immunolabeling was performed on A7r5 cells, renal cryosections, and freshly isolated renal VSMCs with anti-Cav1.2 and anti-Cav2.1 antibodies. Conventional and confocal microscopy revealed expression of both channels in all of the smooth muscle cells. Whole-cell patch clamp on single preglomerular VSMCs from mice showed L-, P-, and Q-type currents. Blockade of the L-type currents by calciseptine (20 nmol/L) inhibited 35.6+/-3.9% of the voltage-dependent Ca2+ current, and blocking P-type currents (omega-agatoxin IVA 10 nmol/L) led to 20.2+/-3.0% inhibition, whereas 300 nmol/L of omega agatoxin IVA (blocking P/Q-type) inhibited 45.0+/-7.3%. In rat aortic smooth muscle cells (A7r5), blockade of L-type channels resulted in 28.5+/-6.1% inhibition, simultaneous blockade of L-type and P-type channels inhibited 58.0+/-11.8%, and simultaneous inhibition of L-, P-, and Q-type channels led to blockade (88.7+/-5.6%) of the Ca2+ current. We conclude that aortic and renal preglomerular smooth muscle cells express L-, P-, and Q-type voltage-dependent Ca2+ channels in the rat and mouse.

摘要

电压依赖性钙通道Cav1.2(L型)和Cav2.1(P/Q型)在血管平滑肌细胞(VSMC)中表达,对肾阻力血管的收缩很重要。在本研究中,我们检测了天然肾VSMC是否共表达L型、P型和Q型钙电流。通过RT-PCR在大鼠和小鼠的肾球前血管中证实了Cav2.1a(P型)和Cav2.1b(Q型)mRNA的表达。用抗Cav1.2和抗Cav2.1抗体对A7r5细胞、肾冰冻切片和新鲜分离的肾VSMC进行免疫标记。传统显微镜和共聚焦显微镜显示,所有平滑肌细胞中均有这两种通道的表达。对小鼠单个球前VSMC进行全细胞膜片钳记录显示存在L型、P型和Q型电流。用钙抑素(20 nmol/L)阻断L型电流可抑制35.6±3.9%的电压依赖性钙电流,阻断P型电流(ω-芋螺毒素IVA 10 nmol/L)导致20.2±3.0%的抑制,而300 nmol/L的ω-芋螺毒素IVA(阻断P/Q型)抑制45.0±7.3%。在大鼠主动脉平滑肌细胞(A7r5)中,阻断L型通道导致28.5±6.1%的抑制,同时阻断L型和P型通道抑制58.0±11.8%,同时抑制L型、P型和Q型通道导致钙电流被阻断(88.7±5.6%)。我们得出结论,大鼠和小鼠的主动脉和肾球前平滑肌细胞表达L型、P型和Q型电压依赖性钙通道。

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