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大鼠附睾中毒蕈碱型乙酰胆碱受体的特性研究

Characterization of muscarinic acetylcholine receptors in the rat epididymis.

作者信息

Maróstica E, Guaze E F, Avellar M C, Porto C S

机构信息

Section of Experimental Endocrinology, Department of Pharmacology, Universidade Federal de São Paulo, Escola Paulista de Medicina, São Paulo, SP 04044-020, Brazil.

出版信息

Biol Reprod. 2001 Oct;65(4):1120-6. doi: 10.1095/biolreprod65.4.1120.

Abstract

The aim of the present study was to characterize the muscarinic acetylcholine receptor subtypes present in the caput and cauda of rat epididymis. The specific binding of [3H]quinuclidinyl benzilate ([3H]QNB) to epididymal membranes was time dependent, temperature dependent, and saturable. The cauda epididymis showed higher affinity to [3H]QNB and higher muscarinic receptor density when compared to the caput region. The [3H]QNB binding was tested in competition studies with different muscarinic receptor antagonists. Each antagonist tested displaced [3H]QNB bound to caput and cauda epididymal membrane with similar affinity. Correlation among the negative logarithm of inhibition constant values (pK(i)) for these antagonists obtained in the epididymis with their correspondent published pK(i) values obtained in tissues that expressed each receptor subtype (M1, M2, M3, and M4) indicated that the muscarinic receptors present in caput and cauda epididymis belong to the muscarinic M2 receptor subtype. When reverse transcription-polymerase chain reaction was used to identify muscarinic receptor mRNA subtypes in the epididymis, only m2 transcripts were detected in the caput region, while both m2 and m3 mRNA subtypes were observed in the cauda region. In conclusion, these results demonstrate that muscarinic receptors are present in the rat epididymis, with expression levels dependent on the region of the epididymis analyzed. Thus, the cholinergic neurotransmitter in the epididymis may be a factor controlling contractility and/or the luminal fluid microenvironment.

摘要

本研究的目的是鉴定大鼠附睾头和附睾尾中存在的毒蕈碱型乙酰胆碱受体亚型。[3H]喹核醇基苯甲酸酯([3H]QNB)与附睾膜的特异性结合具有时间依赖性、温度依赖性且可饱和。与附睾头区域相比,附睾尾对[3H]QNB的亲和力更高,毒蕈碱受体密度也更高。在与不同毒蕈碱受体拮抗剂的竞争研究中测试了[3H]QNB结合情况。所测试的每种拮抗剂都以相似的亲和力取代了与附睾头和附睾尾膜结合的[3H]QNB。在附睾中获得的这些拮抗剂的抑制常数负对数值(pK(i))与其在表达每种受体亚型(M1、M2、M3和M4)的组织中公布的相应pK(i)值之间的相关性表明,附睾头和附睾尾中存在的毒蕈碱受体属于毒蕈碱M2受体亚型。当使用逆转录-聚合酶链反应来鉴定附睾中的毒蕈碱受体mRNA亚型时,在附睾头区域仅检测到m2转录本,而在附睾尾区域观察到m2和m3 mRNA亚型。总之,这些结果表明毒蕈碱受体存在于大鼠附睾中,其表达水平取决于所分析的附睾区域。因此,附睾中的胆碱能神经递质可能是控制收缩性和/或管腔液微环境的一个因素。

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