Synaptogenesis in purified cortical subplate neurons.
Synaptogenesis in purified cortical subplate neurons.
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纯化的皮质亚板神经元中的突触发生。
DOI：
10.1093/cercor/bhn194
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发表时间：
2009
期刊：
影响因子：
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通讯作者：
Shatz,CarlaJ
中科院分区：
文献类型：
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作者：
McKellar,ClaireE;Shatz,CarlaJ
研究突触发生过程中的事件的理想准备是突触稀疏，但可以通过快速的外源性触发随意诱导。我们描述了一种免疫纯化的亚板神经元的培养系统，在该系统中可以触发突触发生，为突触发育的研究提供了新皮层神经元的第一个同质培养。免疫纯化的大鼠亚板神经元中的突触是稀疏的，并且可以通过48小时暴露于神经元和胶质细胞的饲养层来诱导，诱导比任何先前报道的更快。诱导的突触是电生理功能和超微结构正常。微阵列和实时PCR实验揭示了伴随突触发生的基因表达的新程序。令人惊讶的是，很少有已知的突触基因在突触发生的前24小时上调;基因本体注释揭示了突触基因仅在稍后的时间优先上调。原位杂交证实，在文化中调节的一些基因也表达在发育中的皮质。该培养系统提供了一种在同质的皮质神经元群体中研究突触形成的方法，以及更好的突触发生同步化，允许在触发突触形成后调查神经元范围内的事件。
An ideal preparation for investigating events during synaptogenesis would be one in which synapses are sparse, but can be induced at will using a rapid, exogenous trigger. We describe a culture system of immunopurified subplate neurons in which synaptogenesis can be triggered, providing the first homogeneous culture of neocortical neurons for the investigation of synapse development. Synapses in immunopurified rat subplate neurons are sparse, and can be induced by a 48-h exposure to feeder layers of neurons and glia, an induction more rapid than any previously reported. Induced synapses are electrophysiologically functional and ultrastructurally normal. Microarray and real-time PCR experiments reveal a new program of gene expression accompanying synaptogenesis. Surprisingly few known synaptic genes are upregulated during the first 24 h of synaptogenesis; Gene Ontology annotation reveals a preferential upregulation of synaptic genes only at a later time. In situ hybridization confirms that some of the genes regulated in cultures are also expressed in the developing cortex. This culture system provides both a means of studying synapse formation in a homogeneous population of cortical neurons, and better synchronization of synaptogenesis, permitting the investigation of neuron-wide events following the triggering of synapse formation.