Wang Xin, Piccolo Chiara W, Cohen Bruce M, Buttner Edgar A
Department of Psychiatry, Harvard Medical School , Boston, Massachusetts , USA.
J Neurogenet. 2014 Mar-Jun;28(1-2):86-97. doi: 10.3109/01677063.2013.879717. Epub 2014 Feb 25.
The molecular mechanisms of action of antipsychotic drugs (APDs) are not fully understood. Here, we characterize phenotypes of missense and knockout mutations in the Caenorhabditis elegans transient receptor potential melastatin (TRPM) channel ortholog gtl-2, a candidate APD target identified in a genome-wide RNAi (RNA interference) screen for Suppressors of Clozapine-induced Larval Arrest (scla genes). We then employ the developmental phenotypes of gtl-2(lf) mutants to validate our previous gtl-2(RNAi) result. GTL-2 acts in the excretory canal cell to regulate Mg(2+) homeostasis. Using exc (excretory canal abnormal) gene mutants, we demonstrate that excretory canal cell function is necessary for clozapine-induced developmental delay and lethality. Moreover, cell-specific promoter-driven expression studies reveal that GTL-2 function in the excretory canal cell is important for its role in the SCLA phenotype. We then investigate the mechanism by which GTL-2 function in the excretory canal cell impacts clozapine-induced phenotypes. gtl-2(lf) mutations cause hypermagnesemia, and we show that exposure of the wild-type strain to high Mg(2+) phenocopies gtl-2(lf) with respect to suppression of clozapine-induced developmental delay and lethality. Our results suggest that GTL-2 TRPM channel function in the excretory canal cell is important for clozapine's developmental effects. TRP channels are expressed in mammalian brain and are implicated in the pathogenesis of mental illnesses but have not been previously implicated in APD action.
抗精神病药物(APDs)的分子作用机制尚未完全明确。在此,我们对秀丽隐杆线虫瞬时受体电位香草酸亚型(TRPM)通道直系同源基因gtl-2中的错义突变和敲除突变的表型进行了表征,gtl-2是在全基因组RNA干扰(RNAi)筛选氯氮平诱导幼虫停滞抑制因子(scla基因)中鉴定出的一个APD候选靶点。然后,我们利用gtl-2(功能缺失)突变体的发育表型来验证我们之前的gtl-2(RNAi)结果。GTL-2在排泄管细胞中发挥作用,以调节镁离子(Mg²⁺)稳态。利用exc(排泄管异常)基因突变体,我们证明排泄管细胞功能对于氯氮平诱导的发育延迟和致死性是必需的。此外,细胞特异性启动子驱动的表达研究表明,GTL-2在排泄管细胞中的功能对于其在SCLA表型中的作用很重要。接着,我们研究了GTL-2在排泄管细胞中的功能影响氯氮平诱导表型的机制。gtl-2(功能缺失)突变会导致高镁血症,并且我们表明,野生型菌株暴露于高镁环境会在抑制氯氮平诱导的发育延迟和致死性方面模拟gtl-2(功能缺失)的表型。我们的结果表明,排泄管细胞中的GTL-2 TRPM通道功能对于氯氮平的发育效应很重要。TRP通道在哺乳动物大脑中表达,并且与精神疾病的发病机制有关,但此前尚未涉及APD的作用。