Time-lapse in vivo imaging of dorsal root nerve regeneration in mice.
Time-lapse in vivo imaging of dorsal root nerve regeneration in mice.
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小鼠背根神经再生的延时体内成像。
DOI：
10.1007/978-1-4939-0777-9_18
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Son,Young-Jin
中科院分区：
文献类型：
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作者：
Skuba,Andrew;Manire,MeredithAnn;Kim,Hyukmin;Han,SeungBaek;Son,Young-Jin
原发性感觉轴突损伤在脊髓和神经根损伤后很常见，并导致患者遭受慢性疼痛和感觉和运动协调的持续丧失。这种损伤的破坏性后果主要是由于切断的轴突在受损的CNS内再生失败。我们对在阻止或促进功能再生中起关键作用的分子和细胞事件的理解还远远没有完成，部分原因是与神经损伤相关的复杂和动态变化必须从多个动物死后获得的静态图像的比较中推断出来。光学和小鼠转基因技术的革命性创新现在允许直接在活体动物中实时监测再生背根轴突。在这里，我们描述了详细的程序，重复监测确定的轴突在腰椎背根数小时至数周内使用宽视场和双光子显微镜。我们还讨论了体内成像的优势和局限性，并根据我们自己的经验提供了与重复麻醉，广泛椎板切除术和术后护理相关的故障排除问题的建议。这些技术提供了前所未有的机会，以获得新的见解，为什么感觉轴突不能重新进入脊髓。
Primary sensory axon injury is common after spinal cord and root injuries and causes patients to suffer chronic pain and persistent loss of sensation and motor coordination. The devastating consequences of such injuries are due primarily to the failure of severed axons to regenerate within the damaged CNS. Our understanding of the molecular and cellular events that play key roles in preventing or promoting functional regeneration is far from complete, in part because complex and dynamic changes associated with nerve injury have had to be deduced from comparisons of static images obtained from multiple animals after their death. Revolutionary innovations in optics and mouse transgenics now permit real-time monitoring of regenerating dorsal root axons directly in living animals. Here, we describe detailed procedures for repetitive monitoring of identified axons in a lumbar dorsal root over hours to weeks using both widefield and two-photon microscopes. We also discuss the strengths and limitations of in vivo imaging and provide suggestions based on our own experience for troubleshooting issues associated with repeated anesthetization, an extensive laminectomy, and post-op care. These techniques provide the unprecedented opportunity to obtain novel insights into why sensory axons fail to reenter the spinal cord.