Department of Cell Biology & Anatomy, Hotchkiss Brain Institute and Alberta Children's Hospital Research Institute, University of Calgary, Calgary, Alberta, T2N 1N4, Canada.
Department of Neuroscience, Hotchkiss Brain Institute, University of Calgary, Calgary, Alberta, T2N 1N4, Canada.
Sci Rep. 2017 May 11;7(1):1768. doi: 10.1038/s41598-017-01825-x.
In the central nervous system (CNS), cholinergic transmission induces synaptic plasticity that is required for learning and memory. However, our understanding of the development and maintenance of cholinergic circuits is limited, as the factors regulating the expression and clustering of neuronal nicotinic acetylcholine receptors (nAChRs) remain poorly defined. Recent studies from our group have implicated calpain-dependent proteolytic fragments of menin, the product of the MEN1 tumor suppressor gene, in coordinating the transcription and synaptic clustering of nAChRs in invertebrate central neurons. Here, we sought to determine whether an analogous cholinergic mechanism underlies menin's synaptogenic function in the vertebrate CNS. Our data from mouse primary hippocampal cultures demonstrate that menin and its calpain-dependent C-terminal fragment (C-menin) regulate the subunit-specific transcription and synaptic clustering of neuronal nAChRs, respectively. MEN1 knockdown decreased nAChR α5 subunit expression, the clustering of α7 subunit-containing nAChRs at glutamatergic presynaptic terminals, and nicotine-induced presynaptic facilitation. Moreover, the number and function of glutamatergic synapses was unaffected by MEN1 knockdown, indicating that the synaptogenic actions of menin are specific to cholinergic regulation. Taken together, our results suggest that the influence of menin on synapse formation and synaptic plasticity occur via modulation of nAChR channel subunit composition and functional clustering.
在中枢神经系统 (CNS) 中,胆碱能传递诱导了突触可塑性,这是学习和记忆所必需的。然而,我们对胆碱能回路的发育和维持的理解是有限的,因为调节神经元烟碱型乙酰胆碱受体 (nAChRs) 表达和聚集的因素仍未得到很好的定义。我们小组的最近研究表明, MEN1 肿瘤抑制基因产物 menin 的钙蛋白酶依赖性蛋白水解片段参与协调无脊椎动物中枢神经元中 nAChRs 的转录和突触聚集。在这里,我们试图确定 MEN1 在脊椎动物 CNS 中的促突触形成功能是否基于类似的胆碱能机制。我们从小鼠原代海马培养物中获得的数据表明,menin 和其钙蛋白酶依赖性 C 端片段 (C-menin) 分别调节神经元 nAChRs 的亚基特异性转录和突触聚集。 MEN1 敲低降低了 nAChR α5 亚基的表达、α7 亚基组成的 nAChRs 在谷氨酸能突触前末端的聚集以及尼古丁诱导的突触前易化。此外,MEN1 敲低对谷氨酸能突触的数量和功能没有影响,这表明 menin 的促突触作用是特异性的胆碱能调节。总之,我们的研究结果表明,menin 对突触形成和突触可塑性的影响是通过调节 nAChR 通道亚基组成和功能聚集来实现的。