Fu Zhanyan, Washbourne Philip, Ortinski Pavel, Vicini Stefano
Department of Physiology and Biophysics, Georgetown University School of Medicine, Washington, DC 20007, USA.
J Neurophysiol. 2003 Dec;90(6):3950-7. doi: 10.1152/jn.00647.2003. Epub 2003 Aug 20.
The discovery that neuroligin is a key protein involved in synapse formation offers the unprecedented opportunity to induce functional synapses between neurons and heterologous cells. We took this opportunity recording for the first-time synaptic currents in human embryonic kidney 293 (HEK293) cells transfected with neuroligin and the N-methyl-d-aspartate or AMPA receptor subunits in a co-culture with rat cerebellar granule cells. These currents were similar to synaptic currents recorded in neurons, and their decay kinetics was determined by the postsynaptic subunit combination. Although neuroligin expression was sufficient to detect functional synapses, cotransfection of HEK293 cells with Postsynaptic density-95/synapse-associated protein-90 (PSD-95) significantly increased current frequency. Our results support the central role of neuroligin in the formation of CNS synapses, validate the proposal that PSD-95 allows synaptic maturation, and provide a unique experimental model to study how molecular components determine functional properties of excitatory synapses.
发现神经连接蛋白是参与突触形成的关键蛋白质，这为诱导神经元与异源细胞之间形成功能性突触提供了前所未有的机会。我们利用这个机会，首次在与大鼠小脑颗粒细胞共培养的、转染了神经连接蛋白以及N-甲基-D-天冬氨酸或α-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体亚基的人胚肾293（HEK293）细胞中记录突触电流。这些电流与在神经元中记录到的突触电流相似，其衰减动力学由突触后亚基组合决定。虽然神经连接蛋白的表达足以检测到功能性突触，但将HEK293细胞与突触后致密蛋白95/突触相关蛋白90（PSD-95）共转染可显著增加电流频率。我们的结果支持神经连接蛋白在中枢神经系统突触形成中的核心作用，验证了PSD-95促进突触成熟的观点，并提供了一个独特的实验模型来研究分子成分如何决定兴奋性突触的功能特性。
