据报道,G蛋白偶联受体158(GPR158)的异常表达与影响中枢神经系统的多种疾病密不可分。包括阿尔茨海默病(AD),抑郁症,眼内压,还有神经胶质瘤,但是由于缺乏生物学和药理学工具来阐述其优先的细胞分布和分子相互作用网络,其潜在机制仍然难以捉摸。为了评估细胞定位,表达式,和GPR158的功能,我们产生了一个表位标记的GPR158小鼠模型(GPR158Tag),表现出正常的运动,认知,和社会行为,社会记忆没有缺陷,与P60时的C57BL/6J对照小鼠相比,没有焦虑样行为。使用免疫荧光,我们发现GPR158+细胞分布在几个脑区,包括大脑皮层,海马体,小脑,和尾状壳核。接下来,以成年GPR158Tag小鼠的大脑皮层为代表区域,我们发现GPR158只在神经元中表达,而不是在小胶质细胞中,少突胶质细胞,或者星形胶质细胞.值得注意的是,GPR158的大部分富集在Camk2a神经元中,而在PV中间神经元中发现了有限的表达。伴随的3D共定位分析显示,GPR158主要分布在突触后膜,但在突触前膜有一小部分。最后,通过质谱分析,我们鉴定了可能与GPR158相互作用的蛋白质,相关富集途径与免疫荧光研究结果一致.对GPR158-/-小鼠大脑皮层的RNA-seq分析表明,GPR158及其推定的相互作用蛋白参与了氯化物通道复合物和突触小泡膜的组成。使用这些GPR158Tag小鼠,我们能够准确标记GPR158并揭示其在突触小泡功能和记忆中的基本功能。因此,这个模型将是一个有用的工具,用于随后的生物,药理学,和与GPR158相关的电生理研究。
Aberrant expression of G-protein-coupled receptor 158 (
GPR158) has been reported to be inextricably linked to a variety of diseases affecting the central nervous system, including Alzheimer\'s disease (AD), depression, intraocular pressure, and glioma, but the underlying mechanism remains elusive due to a lack of biological and pharmacological tools to elaborate its preferential cellular distribution and molecular interaction network. To assess the cellular localization, expression, and function of
GPR158, we generated an epitope-tagged GPR158 mouse model (GPR158Tag) that exhibited normal motor, cognitive, and social behavior, no deficiencies in social memory, and no anxiety-like behavior compared to C57BL/6J control mice at P60. Using immunofluorescence, we found that GPR158+ cells were distributed in several brain regions including the cerebral cortex, hippocampus, cerebellum, and caudate putamen. Next, using the cerebral cortex of the adult GPR158Tag mice as a representative region, we found that GPR158 was only expressed in neurons, and not in microglia, oligodendrocytes, or astrocytes. Remarkably, the majority of
GPR158 was enriched in Camk2a+ neurons whilst limited expression was found in PV+ interneurons. Concomitant 3D co-localization analysis revealed that GPR158 was mainly distributed in the postsynaptic membrane, but with a small portion in the presynaptic membrane. Lastly, via mass spectrometry analysis, we identified proteins that may interact with
GPR158, and the relevant enrichment pathways were consistent with the immunofluorescence findings. RNA-seq analysis of the cerebral cortex of the
GPR158-/- mice showed that
GPR158 and its putative interacting proteins are involved in the chloride channel complex and synaptic vesicle membrane composition. Using these GPR158Tag mice, we were able to accurately label GPR158 and uncover its fundamental function in synaptic vesicle function and memory. Thus, this model will be a useful tool for subsequent biological, pharmacological, and electrophysiological studies related to GPR158.