Department of Pharmacology, Cambridge, UK.
Br J Pharmacol. 2010 Aug;160(8):1953-62. doi: 10.1111/j.1476-5381.2010.00763.x.
P2Y receptors evoke Ca(2+) signals in vascular smooth muscle cells and regulate contraction and proliferation, but the roles of the different P2Y receptor subtypes are incompletely resolved.
Quantitative PCR was used to define expression of mRNA encoding P2Y receptor subtypes in freshly isolated and cultured rat aortic smooth muscle cells (ASMC). Fluorescent indicators in combination with selective ligands were used to measure the changes in cytosolic free [Ca(2+)] in cultured ASMC evoked by each P2Y receptor subtype.
The mRNA for all rat P2Y receptor subtypes are expressed at various levels in cultured ASMC. Four P2Y receptor subtypes (P2Y1, P2Y2, P2Y4 and P2Y6) evoke Ca(2+) signals that require activation of phospholipase C and comprise both release of Ca(2+) from stores and Ca(2+) entry across the plasma membrane.
Combining analysis of P2Y receptor expression with functional analyses using selective agonists and antagonists, we isolated the Ca(2+) signals evoked in ASMC by activation of P2Y1, P2Y2, P2Y4 and P2Y6 receptors.
P2Y 受体在血管平滑肌细胞中引发 Ca(2+)信号,并调节收缩和增殖,但不同 P2Y 受体亚型的作用尚未完全明确。
使用定量 PCR 方法确定新分离和培养的大鼠主动脉平滑肌细胞(ASMC)中编码 P2Y 受体亚型的 mRNA 的表达。使用荧光指示剂结合选择性配体来测量每种 P2Y 受体亚型在培养的 ASMC 中引发的细胞浆游离 Ca(2+)的变化。
所有大鼠 P2Y 受体亚型的 mRNA 在培养的 ASMC 中以不同水平表达。四种 P2Y 受体亚型(P2Y1、P2Y2、P2Y4 和 P2Y6)引发 Ca(2+)信号,需要激活磷脂酶 C,包括从储存库中释放 Ca(2+)和穿过质膜的 Ca(2+)内流。
我们结合使用选择性激动剂和拮抗剂的 P2Y 受体表达分析和功能分析,分离了 P2Y1、P2Y2、P2Y4 和 P2Y6 受体激活在 ASMC 中引发的 Ca(2+)信号。