Cloning and functional comparison of kappa and delta opioid receptors from mouse brain.
Cloning and functional comparison of kappa and delta opioid receptors from mouse brain.
复制标题
DOI：
10.1073/pnas.90.14.6736
复制
发表时间：
1993-07
影响因子：
11.1
通讯作者：
K. Yasuda;K. Raynor;H. Kong;C. Breder;J. Takeda;T. Reisine;G. Bell
中科院分区：
文献类型：
--
作者：
K. Yasuda;K. Raynor;H. Kong;C. Breder;J. Takeda;T. Reisine;G. Bell
在试图确定生长抑素受体家族的G蛋白偶联受体的新成员，我们分离的cDNA从小鼠脑库编码两个相关的受体样蛋白，指定msl-1和msl-2，分别为380和372个氨基酸。msl-1和msl-2的同源性分别为61%和71%。在G蛋白偶联受体超家族的成员中，msl-1和msl-1的序列与生长抑素受体（SSTR）的序列最密切相关，与SSTR 1的序列具有约35%的同一性。在COS-1细胞中的瞬时表达表明msl-1和msl-2不与生长抑素结合。相反，它们选择性地和高亲和力地结合阿片类药物，并分别具有κ和δ阿片受体的药理学特性。事实上，msl-2的序列与其他研究人员最近克隆的δ阿片受体的序列完全相同。kappa/msl-1和delta/msl-2阿片受体的功能表征表明，它们与G蛋白偶联，并介导阿片受体类特异性激动剂抑制毛喉素刺激的cAMP形成。RNA印迹和原位杂交组织化学研究表明，κ阿片受体mRNA在大脑中的新皮层，海马，杏仁核，内侧缰核，下丘脑（弓状核和室旁核），蓝斑，臂旁核的高水平表达，这表明该受体可能在唤醒和调节自主神经和神经内分泌功能中发挥作用。
While trying to identify new members of the somatostatin receptor family of G protein-coupled receptors, we isolated cDNAs from a mouse brain library encoding two related receptor-like proteins, designated msl-1 and msl-2, of 380 and 372 amino acids, respectively. There was 61% identity and 71% similarity between the sequences of msl-1 and msl-2. Among members of the G protein-coupled receptor superfamily, the sequences of both msl-1 and msl-1 were most closely related to those of the somatostatin receptors (SSTRs), having approximately 35% identity with the sequence of SSTR1. Transient expression in COS-1 cells showed that msl-1 and msl-2 did not bind somatostatin. Rather they bound opioids selectively and with high affinity and had the pharmacological properties of kappa and delta opioid receptors, respectively. Indeed, the sequence of msl-2 was identical to that of a delta opioid receptor recently cloned by other workers. Functional characterization of kappa/msl-1 and delta/msl-2 opioid receptors showed that they were coupled to G proteins and mediated opioid receptor class-specific agonist inhibition of forskolin-stimulated cAMP formation. RNA blotting studies and in situ hybridization histochemistry showed that kappa opioid receptor mRNA was expressed at high levels in brain in the neocortex, hippocampus, amygdala, medial habenula, hypothalamus (arcuate and paraventricular nuclei), locus ceruleus, and parabrachial nucleus, suggesting that this receptor may play a role in arousal and regulation of autonomic and neuroendocrine functions.