Cloning, pharmacological characterisation and distribution of the rat G-protein-coupled P2Y13 receptor
Cloning, pharmacological characterisation and distribution of the rat G-protein-coupled P2Y13 receptor
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DOI：
10.1016/j.bcp.2004.02.038
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发表时间：
2004-07-01
影响因子：
5.8
通讯作者：
Abbracchio, MP
中科院分区：
文献类型：
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作者：
Fumagalli, M;Trincavelli, L;Abbracchio, MP
人P2Y(13)受体是一种新的受体，其特征是偶联Gi，对腺嘌呤二磷核苷酸有反应性，并被P2Y拮抗剂AR-C69931MX阻断。小鼠的P2Y(13)直系同源基因也已被报道。本文首次报道了大鼠P2Y(13)受体的克隆、药理分析和组织分布。大鼠P2Y(13)与人和小鼠的P2Y(13)受体分别有79%和87%的同源性。在1321N1细胞中表达rP2Y(13)受体可诱导对典型的P2Y(13)受体激动剂ADP和2MeSADP的反应，这可通过刺激[S-35]GTPGammaS结合检测到。在存在Galpha(16)亚单位的情况下，转染rP2Y(13)受体的细胞具有更高的激动剂活性；在所有情况下，激动剂作用都被百日咳毒素预处理所消除。在与人和小鼠的受体不同时，ADP比2MeSADP更有效。其他核苷酸和糖核苷酸无效。在没有Galpha(16)和存在Galpha(16)的情况下，ADP和2MeSADP对rP2Y(13)受体的激活均被NM浓度的AR-C69931MX完全抑制。相反，选择性的P2Y(1)受体拮抗剂MRS2179对rP2Y(13)受体无抑制作用。RP2Y(13)受体在脾中的表达水平最高，其次是肝脏和大脑(据报道，在大脑皮质和纹状体中表达水平特别高)，这表明rP2Y(13)受体在神经系统和免疫系统中发挥着重要作用。在原代培养的大鼠星形胶质细胞中发现了与其他克隆的P2Y受体相似的表达水平，这表明可能在反应性星形胶质细胞增生症中起作用。因此，大鼠P2Y(13)受体与其人和小鼠的同源物显示出一些相似之处，但也有一些有趣的不同之处，当在大鼠动物模型中表征该受体的病理生理作用时，必须考虑到这一点。(C)2004 Elsevier Inc.保留所有权利。
The human P2Y(13) receptor is a new receptor characterized by coupling to Gi, responsiveness to adenine di-phospho-nucleotides and blockade by the P2Y antagonist AR-C69931MX. The mouse P2Y(13) ortholog has also been reported. Here we report, for the first time, the cloning of rat P2Y(13) receptor, its pharmacological analysis and tissue distribution. Rat P2Y(13) is 79% and 87% identical to human and mouse P2Y(13) receptors, respectively. Expression of rP2Y(13) receptor in 1321N1 cells induced the appearance of responses to the typical P2Y(13) receptor agonists ADP and 2MeSADP, as detected by stimulation of [S-35]GTPgammaS binding. Agonist activities were higher in cells transfected with rP2Y(13) receptor in the presence of the Galpha(16) subunit; in all cases agonist effects were abolished by pertussis toxin pretreatment. At variance from both human and mouse receptors, ADP was more potent than 2MeSADP. Other nucleotides and sugar-nucleotides were ineffective. Both in the absence and presence of Galpha(16), activation of rP2Y(13) receptor by ADP and 2MeSADP was completely inhibited by nM concentrations of AR-C69931MX. In contrast, no inhibition of rP2Y(13) receptor was induced by the selective P2Y(1) receptor antagonist MRS2179. rP2Y(13) receptor showed highest expression levels in spleen, followed by liver and brain (with particularly high levels in cortex and striatum as reported in man), suggesting important roles in the nervous and immune systems. Expression levels comparable to those of the other cloned P2Y receptors were found in primary rat astrocytes, indicating a possible role in reactive astrogliosis. Hence, rat P2Y(13) receptor displays several similarities but also interesting differences with its human and mouse orthologs, that will have to be taken into account when characterizing the pathophysiological roles of this receptor in the rat animal models. (C) 2004 Elsevier Inc. All rights reserved.