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秀丽隐杆线虫D2样多巴胺受体的克隆与鉴定

Cloning and characterization of a Caenorhabditis elegans D2-like dopamine receptor.

作者信息

Suo Satoshi, Sasagawa Noboru, Ishiura Shoichi

机构信息

Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Tokyo, Japan.

出版信息

J Neurochem. 2003 Aug;86(4):869-78. doi: 10.1046/j.1471-4159.2003.01896.x.

Abstract

The neurotransmitter dopamine plays an important role in the regulation of behavior in both vertebrates and invertebrates. In mammals, dopamine binds and activates two classes of dopamine receptors, D1-like and D2-like receptors. However, D2-like dopamine receptors in Caenorhabditis elegans have not yet been characterized. We have cloned a cDNA encoding a putative C. elegans D2-like dopamine receptor. The deduced amino acid sequence of the cloned cDNA shows higher sequence similarities to vertebrate D2-like dopamine receptors than to D1-like receptors. Two splice variants that differ in the length of their predicted third intracellular loops were identified. The receptor heterologously expressed in cultured cells showed high affinity binding to [125I]iodo-lysergic acid diethylamide. Dopamine showed the highest affinity for this receptor among several amine neurotransmitters tested. Activation of the heterologously expressed receptor led to the inhibition of cyclic AMP production, confirming that this receptor has the functional property of a D2-like receptor. We have also analyzed the expression pattern of this receptor and found that the receptor is expressed in several neurons including all the dopaminergic neurons in C. elegans.

摘要

神经递质多巴胺在脊椎动物和无脊椎动物的行为调节中都起着重要作用。在哺乳动物中,多巴胺结合并激活两类多巴胺受体,即D1样受体和D2样受体。然而,秀丽隐杆线虫中的D2样多巴胺受体尚未得到表征。我们克隆了一个编码假定的秀丽隐杆线虫D2样多巴胺受体的cDNA。克隆的cDNA推导的氨基酸序列与脊椎动物D2样多巴胺受体的序列相似性高于与D1样受体的相似性。鉴定出了两个预测的第三细胞内环长度不同的剪接变体。在培养细胞中异源表达的受体显示出对[125I]碘-麦角酸二乙酰胺的高亲和力结合。在测试的几种胺类神经递质中,多巴胺对该受体的亲和力最高。异源表达受体的激活导致环磷酸腺苷产生的抑制,证实该受体具有D2样受体的功能特性。我们还分析了该受体的表达模式,发现该受体在包括秀丽隐杆线虫所有多巴胺能神经元在内的几个神经元中表达。

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