The intracellular pathway of the acetylcholine-induced contraction in cat detrusor muscle cells
The intracellular pathway of the acetylcholine-induced contraction in cat detrusor muscle cells
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DOI：
10.1038/sj.bjp.0704954
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发表时间：
2002-12-01
影响因子：
7.3
通讯作者：
Sohn, UD
中科院分区：
文献类型：
--
作者：
An, JY;Yun, HS;Sohn, UD
1 本研究旨在研究涉及乙酰胆碱 (ACh) 诱导的猫逼尿肌细胞收缩的细胞内途径2 收缩表示为通过酶消化获得的单独分离的平滑肌细胞的长度缩短百分比。制备分散的完整细胞和透化细胞，分别用于药物和酶抗体的处理。使用蛋白质印迹，我们证实了相关蛋白的存在。3 ACh 的最大收缩发生在 10(-11) M。这种反应优先被 M-3 毒蕈碱受体拮抗剂 p-氟六氢硅拉非尼醇 (rhoF-HSD) 拮抗，但不会被 M 拮抗剂哌仑西平和 M 毒蕈碱受体拮抗剂甲氧曲明拮抗。我们在膀胱逼尿肌中鉴定出 G 蛋白 G(q/11)、G(s)、G(0)、G(i1)、G(i2) 和 G(i3)。 Gq/11 抗体可选择性抑制 ACh 诱导的收缩，但不会抑制其他 G 亚基。4 磷脂酰肌醇特异性磷脂酶 C (PI-PLC) 抑制剂新霉素可减少 ACh 诱导的收缩。然而，磷脂酶 D、磷脂酶 A 和蛋白激酶 C 的抑制剂不会减弱乙酰胆碱诱导的收缩。 ACh 诱导的收缩被 PLC-β(1) 抗体抑制，但 PLC-β(3) 和 PLC-gamma Thapsigargin 或锶消耗或阻止细胞内钙释放，抑制 ACh 诱导的收缩。肌醇 1,4,5-三磷酸 (IP3) 受体抑制剂肝素可减少 ACh 诱导的收缩。5 这些结果表明，在猫中，ACh 诱导的逼尿肌收缩是通过 M-3 毒蕈碱受体依赖性 G(q/11) 和 PLC-beta(1) 激活以及 IP3 依赖性 Ca2+ 释放介导的。
1 The present study was aimed to investigate intracellular pathways involved in acetylcholine (ACh)-induced contraction in cat detrusor muscle cells2 Contraction was expressed as per cent shortening of length of individually isolated smooth muscle cells obtained by enzymatic digestion. Dispersed intact and permeabilized cells were prepared for the treatment of drugs and antibody to enzymes, respectively. Using Western blot, we confirmed the presence of related proteins.3 The maximal contraction to ACh was generated at 10(-11) M. This response was preferentially antagonized by M-3 muscarinic receptor antagonist p-fluoro-hexahydrosiladifenidol (rhoF-HSD) but not by the M, antagonist pirenzepine and the M, muscarinic receptor antagonist methoctramine. We identified G-proteins G(q/11), G(s), G(0), G(il), G(i2) and G(i3) in the bladder detrusor muscle. ACh-induced contraction was selectively inhibited by Gq/11 antibody but not to other G subunit.4 The phosphatidylinositol-specific phospholipase C (PI-PLC) inhibitor neomycin reduced ACh-induced contraction. However, the inhibitors of the phospholipase D, the phospholipase A, and protein kinase C did not attenuate the ACh-induced contraction. ACh-induced contraction was inhibited by antibody to PLC-beta(1) but not PLC-beta(3) and PLC-gamma Thapsigargin or strontium, which depletes or blocks intracellular calcium release, inhibited ACh-induced contraction. Inositol 1,4,5-triphosphate (IP3) receptor inhibitor heparin reduced ACh-induced contraction.5 These results suggest that in cat detrusor muscle contraction induced by ACh is mediated via M-3 muscarinic receptor-dependent activation of G(q/11) and PLC-beta(1) and IP3-dependent Ca2+ release.