Sevoflurane Impairs Growth Cone Motility in Dissociated Murine Neurons.
Sevoflurane Impairs Growth Cone Motility in Dissociated Murine Neurons.
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DOI：
10.1097/ana.0000000000000360
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发表时间：
2016-10
影响因子：
3.7
通讯作者：
Mintz CD
中科院分区：
文献类型：
--
作者：
Ryu YK;Mathena RP;Lim S;Kwak M;Xu M;Mintz CD
在动物模型中，出生后早期暴露于全身麻醉剂导致学习和记忆的持久损害。解释这一发现的一个假设是，在神经发育的关键点暴露于麻醉剂会破坏大脑回路的形成。在这里，我们探讨了七氟烷对神经元生长锥的影响，神经元生长锥是轴突和树突生长端的一种专门化，负责神经元之间连接的靶向生长。从胚胎小鼠新皮层制备分离的神经元培养物。使用微分干涉对比显微镜拍摄暴露于麻醉剂的活生长锥的时间推移图像，并将板状伪足的面积和丝状伪足尖端的速度的变化率定量为运动性的测量。p75神经营养素受体（p75 NTR）的参与进行了测试，使用抑制剂施加到媒体，并通过免疫共沉淀试验。七氟烷暴露量在2%和6%之间时，轴突和树突生长锥的板状伪足面积变化率和丝状伪足尖端速度均显著降低。运动性可以通过用Y27632和TAT-肽5（其分别是Rho激酶和p75 NTR的抑制剂）处理而基本上恢复。七氟烷导致Rho-GDP解离抑制剂与p75 NTR下拉后的免疫共沉淀减少。七氟烷干扰生长锥运动，这是脑回路形成的关键过程。我们的数据表明，这可能是通过对p75 NTR的作用而发生的，p75 NTR通过Rho途径促进生长抑制信号传导。
Early postnatal exposure to general anesthetic agents causes a lasting impairment in learning and memory in animal models. One hypothesis to explain this finding is that exposure to anesthetic agents during critical points in neural development disrupts the formation of brain circuitry. Here we explore the effects of sevoflurane on the neuronal growth cone, a specialization at the growing end of axons and dendrites that is responsible for the targeted growth that underlies connectivity between neurons. Dissociated neuronal cultures were prepared from embryonic mouse neocortex. Time lapse images of live growth cones exposed to anesthetics were taken using differential interference contrast microscopy, and the rate of change of the area of the lamellipodia and the speed of the filopodial tip were quantified as measures of motility. The involvement of the p75 neurotropin receptor (p75NTR) was tested using inhibitors applied to the media and via a co-immunoprecipitation assay. The rate of lamellipodial area change and filopodial tip velocity in both axonal and dendritic growth cones was significantly reduced with sevoflurane exposure between 2% and 6%. Motility could be substantially restored by treatment with Y27632 and TAT-peptide 5, which are inhibitors of Rho Kinase and p75NTR, respectively. Sevoflurane results in reduced co-immunoprecipitation of Rho-GDP dissociation inhibitor after pulldown with p75NTR. Sevoflurane interferes with growth cone motility, which is a critical process in brain circuitry formation. Our data suggest that this may occur via an action on the p75NTR, which promotes growth inhibitory signaling via the Rho pathway.