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首页> 外文期刊>BMC Neuroscience >Homozygous mutation of focal adhesion kinase in embryonic stem cell derived neurons: normal electrophysiological and morphological properties in vitro
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Homozygous mutation of focal adhesion kinase in embryonic stem cell derived neurons: normal electrophysiological and morphological properties in vitro

机译：胚胎干细胞衍生神经元中粘着斑激酶的纯合突变：体外的正常电生理和形态学特性

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Background Genetically manipulated embryonic stem (ES) cell derived neurons (ESNs) provide a powerful system with which to study the consequences of gene manipulation in mature, synaptically connected neurons in vitro . Here we report a study of focal adhesion kinase (FAK), which has been implicated in synapse formation and regulation of ion channels, using the ESN system to circumvent the embryonic lethality of homozygous FAK mutant mice. Results Mouse ES cells carrying homozygous null mutations (FAK-/-) were generated and differentiated in vitro into neurons. FAK-/- ESNs extended axons and dendrites and formed morphologically and electrophysiologically intact synapses. A detailed study of NMDA receptor gated currents and voltage sensitive calcium currents revealed no difference in their magnitude, or modulation by tyrosine kinases. Conclusion FAK does not have an obligatory role in neuronal differentiation, synapse formation or the expression of NMDA receptor or voltage-gated calcium currents under the conditions used in this study. The use of genetically modified ESNs has great potential for rapidly and effectively examining the consequences of neuronal gene manipulation and is complementary to mouse studies.
机译：背景技术遗传操纵的胚胎干（ES）细胞衍生的神经元（ESN）提供了一个强大的系统，可用来研究基因操纵在体外成熟，突触连接的神经元中的后果。在这里，我们报告了对粘着斑激酶（FAK）的研究，该研究已涉及到突触形成和离子通道的调控，使用ESN系统规避纯合FAK突变小鼠的胚胎致死率。结果产生了携带纯合无效突变（FAK -/-）的小鼠ES细胞，并在体外分化为神经元。 FAK -/- ESNs延伸轴突和树突，并形成形态和电生理完整的突触。对NMDA受体门控电流和电压敏感钙电流的详细研究显示，它们的大小或酪氨酸激酶的调节均无差异。结论在本研究条件下，FAK在神经元分化，突触形成或NMDA受体或电压门控钙电流的表达中没有强制性作用。转基因ESN的使用具有迅速有效地检查神经元基因操纵的后果的巨大潜力，并且与小鼠研究相辅相成。

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