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首页> 美国卫生研究院文献>Cells >Source of Early Regenerating Axons in Lamprey Spinal Cord Revealed by Wholemount Optical Clearing with BABB
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Source of Early Regenerating Axons in Lamprey Spinal Cord Revealed by Wholemount Optical Clearing with BABB

机译：用BABB的Wholemount光学清洁显示Lamprey脊髓早期再生轴突的来源

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Many studies of axon regeneration in the lamprey focus on 18 pairs of large identified reticulospinal (RS) neurons, whose regenerative abilities have been individually quantified. Their axons retract during the first 2 weeks after transection (TX), and many grow back to the site of injury by 4 weeks. However, locomotor movements begin before 4 weeks and the lesion is invaded by axons as early as 2 weeks post-TX. The origins of these early regenerating axons are unknown. Their identification could be facilitated by studies in central nervous system (CNS) wholemounts, particularly if spatial resolution and examination by confocal microscopy were not limited by light scattering. We have used benzyl alcohol/benzyl benzoate (BABB) clearing to enhance the resolution of neuronal perikarya and regenerated axons by confocal microscopy in lamprey CNS wholemounts, and to assess axon regeneration by retrograde and anterograde labeling with fluorescent dye applied to a second TX caudal or rostral to the original lesion, respectively. We found that over 50% of the early regenerating axons belonged to small neurons in the brainstem. Some propriospinal neurons located close to the TX also contributed to early regeneration. The number of early regenerating propriospinal neurons decreased with distance from the original lesion. Descending axons from the brainstem were labeled anterogradely by application of tracer to a second TX close to the spinal–medullary junction. This limited contamination of the data by regenerating spinal axons whose cell bodies are located rostral or caudal to the TX and confirmed the regeneration of many small RS axons as early as 2 weeks post-TX. Compared with the behavior of axotomized giant axons, the early regenerating axons were of small caliber and showed little retraction, probably because they resealed rapidly after injury.
机译：在Lampley中的许多研究对18对大型鉴定的网状（RS）神经元进行了重点，其再生能力已经单独定量。它们的轴突在横衰期（TX）后的前2周内缩回，并且许多人将恢复到损伤部位4周。然而，11周之前的运动运动开始，并且由于TX后2周，轴突侵入了病变。这些早期再生轴突的起源是未知的。通过中枢神经系统（CNS）的研究可以促进其识别，特别是如果通过共聚焦显微镜的空间分辨率和检查不受光散射的限制。我们已经使用苄醇/苄酯（BABB）清洁，通过CAMPREY CNS WHOLEMOUNT中的共聚焦显微镜通过逆行和荧光染料进行透析和荧光染料的轴颈再生来增强神经元治疗和再生轴突的分辨率。用荧光染料施加到第二个TX尾部或罗斯特拉特分别对原始病变。我们发现超过50％的早期再生轴突属于脑干中的小神经元。靠近Tx的一些血管神经元也有助于早期再生。早期再生的ProbrioSto脊髓神经元的数量随着原始病变的距离而降低。通过将示踪剂施加到靠近脊柱髓质连接的第二TX，从脑干中展示来自脑干的下降轴突。这种通过再生脊柱轴身污染数据，其细胞体位于TX的脊髓杆菌或尾部，并在TX后2周确认了许多小型RS轴突的再生。与轴突的巨型轴突的行为相比，早期再生轴突是小口径，并且表现出几乎没有收回，可能是因为它们在损伤后迅速重新密封。

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