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MOLECULAR ANALYSIS OF THE REGULATION OF MUSCARINIC RECEPTOR EXPRESSION AND FUNCTION

机译:肌肉受体表达和功能调控的分子分析

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Several systems are being used to determine the molecular and cellular basis for the regulation of expression and function of the muscarinic receptors. Treatment of chick heart cells in culture results in decreased levels of mRNA encoding the cm2 and cm4 receptors. This probably results from decreased gene transcription which requires concomitant mAChR-mediated inhibition of adenylyl cyclase and mAChR-mediated stimulation of phospholipase C. Site-directed mutagenesis was used to demonstrate that the single tyrosine residue in the carboxyl-terminal cytoplasmic tail of the m2 receptor is involved in agonist-induced downregulation but not sequestration. Activation of heterologous receptors in chick heart cells can also regulate mAChR mRNA levels. A cAMP-regulated luciferase reporter gene has been used to demonstrate that the m4 receptor preferentially couples to G(i) alpha-2 or G(o) alpha over G(i alpha)-1 or G(i) alpha-3 to mediate inhibition of adenylyl cyclase activity. Finally, in order to determine the role of individual receptor subtypes in muscarinic-mediated responses in vivo, we are beginning to use the method of targeted gene disruption by homologous recombination to generate mice deficient in specific receptor subtypes. [References: 10]
机译:几种系统正在用于确定用于调节毒蕈碱受体的表达和功能的分子和细胞基础。处理培养中的雏鸡心脏细胞会导致编码cm2和cm4受体的mRNA水平降低。这可能是由于基因转录降低所致,该基因转录需要同时进行mAChR介导的腺苷酸环化酶抑制和mAChR介导的磷脂酶C刺激。定点诱变用于证明m2受体羧基末端细胞质尾巴中的单个酪氨酸残基与激动剂引起的下调有关,但与螯合无关。雏鸡心脏细胞中异源受体的激活也可以调节mAChR mRNA水平。已使用cAMP调节的萤光素酶报告基因来证明m4受体比G(i alpha)-1或G(i)alpha-3优先与G(i)alpha-2或G(o)alpha偶联以介导抑制腺苷酸环化酶活性。最后,为了确定单个受体亚型在体内毒蕈碱介导的反应中的作用,我们开始使用通过同源重组破坏靶基因的方法来产生缺乏特定受体亚型的小鼠。 [参考:10]

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