ACCELERATED NEURITE GROWTH FROM SPIRAL GANGLION NEURONS EXPOSED TO THE Rho KINASE INHIBITOR H-1152
ACCELERATED NEURITE GROWTH FROM SPIRAL GANGLION NEURONS EXPOSED TO THE Rho KINASE INHIBITOR H-1152
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DOI：
10.1016/j.neuroscience.2010.05.020
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发表时间：
2010-08-25
期刊：
影响因子：
3.3
通讯作者：
Whitlon, D. S.
中科院分区：
文献类型：
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作者：
Lie, M.;Grover, M.;Whitlon, D. S.
当它们的毛细胞突触伙伴死亡时，双极耳蜗螺旋神经节神经元死亡或缩回它们的外周神经纤维。诱导外周神经突再生的努力不得不依赖于螺旋神经节神经突再生机制的有限知识，并且受到进行大量人工神经突生长反应分析的不切实际性的限制。在这里，我们已经使用了解离培养的出生后小鼠螺旋神经节，以评估Rho激酶抑制剂H-1152对神经突生长的影响，并确定实用的自动化高含量分析，用于评估螺旋神经节神经元在体外的神经突长度。在出生后的小鼠螺旋神经节的培养物中，超过95%的神经元以依赖于初始培养基组合物是否含有白血病抑制因子或骨形态发生蛋白4的比率发展双极、单极或无神经突形态。将两种条件下的培养物保持24小时，然后暴露于H-1152 18小时。暴露于H-1152的培养物均未显示出神经元存活率降低或不同神经元形态比例的改变。然而，手动测量，在H-1152的存在下，在两种类型的文化的神经突长度的人口增加。使用Arrayscan VTi成像仪和Cellomics软件的高含量分析证实了培养条件之间神经突长度的等级顺序差异。这些数据表明在Rho激酶的信号传导途径中存在抑制性调节机制，其减缓螺旋神经节神经突的生长。自动化分析证明了使用解离的小鼠螺旋神经节的原代培养物进行调节神经突生长的化学物质、基因或其他因子的大规模筛选的可行性。(C)2010年IBRO。由爱思唯尔有限公司出版。保留所有权利。
Upon the death of their hair cell synaptic partners, bipolar cochlear spiral ganglion neurons either die or retract their peripheral nerve fibers. Efforts to induce the regrowth of the peripheral neurites have had to rely on limited knowledge of the mechanisms underlying spiral ganglion neurite regeneration and have been restricted by the impracticality of undertaking large numbers of manual analyses of neurite growth responses. Here we have used dissociated cultures of postnatal mouse spiral ganglia to assess the effects of the Rho kinase inhibitor H-1152 on neurite growth and to determine the utility of automated high content analysis for evaluating neurite length from spiral ganglion neurons in vitro. In cultures of postnatal mouse spiral ganglion, greater than 95% of the neurons develop bipolar, monopolar or neurite-free morphologies in ratios dependent on whether the initial medium composition contains leukemia inhibitory factor or bone morphogenetic protein 4. Cultures under both conditions were maintained for 24 h, then exposed for 18 h to H-1152. None of the cultures exposed to H-1152 showed decreased neuronal survival or alterations in the ratios of different neuronal morphologies. However, as measured manually, the population of neurite lengths was increased in the presence of H-1152 in both types of cultures. High content analysis using the Arrayscan VTi imager and Cellomics software confirmed the rank order differences in neurite lengths among culture conditions. These data suggest the presence of an inhibitory regulatory mechanism(s) in the signaling pathway of Rho kinase that slows the growth of spiral ganglion neurites. The automated analysis demonstrates the feasibility of using primary cultures of dissociated mouse spiral ganglion for large scale screens of chemicals, genes or other factors that regulate neurite growth. (C) 2010 IBRO. Published by Elsevier Ltd. All rights reserved.