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Molecular pharmacological study for characterization of new identified iropioid receptor subclass

新鉴定的伊罗片受体亚类的分子药理学研究

基本信息

批准号：
16590058
负责人：
SAKURADA Shinobu
金额：
$ 1.86万
依托单位：
Tohoku Pharmaceutical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590058/
关键词：
Opioid Receptor Receptor Expressed Cell Splice Variants Opioid Peptides

项目摘要

With the cells expressed cloned MOR-1, MOR-1A, MOR-1C or MOR-1E, the pharmacological character of these μ-opioid receptor splice variants were investigated. DAMGO, typical μ-opioid receptor agonist, showed high affinity and intrinsic activity in all of these splice variants. Moreover, amidino-TAPA, selective μ-opioid receptor agonist can lead the release of endogenous x-opioid peptides, also showed high intrinsic activity in all of these splice variants. These evidences clearly suggest that these 4 splice variants are not the μ-opioid receptor splice variants lead the release of endogenous κ-opioid peptides. To identify the splice varinats lead the release of endogenous κ-opioid peptides, we decide to identify first the amidino-TAPA-sensitive DAMGO-insensitive splice variants among more than 30 splice variants in behavioral experiment using the exon-specific antisense oligodeoxynucleotides for μ-opioid receptor gene. As the results, MOR-1J, MOR-1K and MOR-1L were identified as the amid … More ino-TAPA-sensitive DAMGO-insensitive splice variants. Moreover, additional experiments using the antisera against endogenous κ-opioid peptides, dynorphin A, dynorphin B or α-neoendorphin, identified that MOR-1J and MOR-1L are splice variants lead the release of dynorphin A, whereas MOR-1K is a splice variant lead the release of dynorphin B and α-neoendorphin. With above evidence, MOR-1J, MOR-1K and MOR-1L were cloned, but no intrinsic activity of amidino-TAPA was observed in the cells expressed cloned MOR-1K and MOR-1L. Since these 2 splice variants do not contain transmembrane structure, these splice variants may not have any function without other splice variants contain 7-transmembrane structure. To identify the pharmacological character of MOR-1J, MOR-1K and MOR-1L in its expressed cell, the functional characterization of heterodimer of these splice variants with other. Μ-opioid receptor splice variants, which contain 7-transmembrane structure, may be required. The cDNA of MOR-1J, MOR-1K and MOR-1L, cloned in the present project, are scheduled to be used in the new project accepted. Less

用表达的细胞克隆MOR-1、MOR-1A、MOR-1C或MOR-1E，研究这些μ-阿片受体剪接变异体的药理特性。DAMGO是典型的μ阿片受体激动剂，在所有这些剪接变异体中都显示出高亲和力和内在活性。此外，选择性的μ-阿片受体激动剂氨基-TAPA可以导致内源性x-阿片肽的释放，在所有这些剪接变异体中也显示出高的内源性活性。这些证据清楚地表明，这4个剪接变异体不是μ-阿片受体剪接变异体，导致内源性κ-阿片肽的释放。为了确定导致内源性κ阿片肽释放的剪接变异体，我们决定在行为实验中首先利用μ阿片受体基因的外显子特异性反义寡核苷酸从30多个剪接变异体中鉴定出氨基-TAPA敏感的DAMGO不敏感的剪接变异体。结果表明，MOR-1J、MOR-1K和MOR-1L为AMID…对TAPA更敏感，对DAMGO不敏感的剪接变体。此外，使用针对内源性κ阿片肽、强啡肽A、强啡肽B或α-新内啡肽的抗血清的进一步实验证实，MOR-1J和MOR-1L是导致强啡肽A释放的剪接变体，而MOR-1K是导致强啡肽B和α-新内啡肽释放的剪接变体。有了上述证据，我们克隆了MOR-1J、MOR-1K和MOR-1L，但在表达MOR-1K和MOR-1L的细胞中没有观察到氨基转移酶的内源性活性。由于这两个剪接变异体不包含跨膜结构，如果没有其他包含7-跨膜结构的剪接变异体，这些剪接变异体可能没有任何功能。为了鉴定MOR-1J、MOR-1K和MOR-1L在其表达细胞中的药理特性，对这些剪接变异体与其他异源二聚体的功能进行了鉴定。可能需要包含7-跨膜结构的Μ-阿片受体剪接变体。本项目中克隆的MOR-1J、MOR-1K和MOR-1L的基因，计划用于新项目的验收。较少