Three-dimensional imaging of the unsectioned adult spinal cord to assess axon regeneration and glial responses after injury
Three-dimensional imaging of the unsectioned adult spinal cord to assess axon regeneration and glial responses after injury
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DOI：
10.1038/nm.2600
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发表时间：
2012-01-01
期刊：
影响因子：
82.9
通讯作者：
Bradke, Frank
中科院分区：
文献类型：
--
作者：
Ertuerk, Ali;Mauch, Christoph P.;Bradke, Frank
目前的组织学和成像技术阻碍了中枢神经系统（CNS）再生的研究，因为它们只能提供轴突和神经胶质反应的部分信息。在这里，我们开发了一种基于四氢呋喃的清除程序，使固定和未切片的成人中枢神经系统组织透明和完全可穿透的光学成像。在大的脊髓节段，我们成像荧光标记的细胞的“超微”和双光子显微镜，而不需要组织切片。我们发现，超过一年后，受伤的生长能力轴突再生丰富的损伤部位。一些生长不全的轴突也可以绕过病变再生。此外，我们准确地确定了脊髓损伤后胶质细胞的定量变化。因此，清除CNS组织能够明确评估轴突再生和神经胶质反应。我们的清除程序也使其他器官透明，这使得这种方法可用于大量的临床前范例。
Studying regeneration in the central nervous system (CNS) is hampered by current histological and imaging techniques because they provide only partial information about axonal and glial reactions. Here we developed a tetrahydrofuranbased clearing procedure that renders fixed and unsectioned adult CNS tissue transparent and fully penetrable for optical imaging. In large spinal cord segments, we imaged fluorescently labeled cells by `ultramicroscopy' and two-photon microscopy without the need for histological sectioning. We found that more than a year after injury growth-competent axons regenerated abundantly through the injury site. A few growth-incompetent axons could also regenerate when they bypassed the lesion. Moreover, we accurately determined quantitative changes of glial cells after spinal cord injury. Thus, clearing CNS tissue enables an unambiguous evaluation of axon regeneration and glial reactions. Our clearing procedure also renders other organs transparent, which makes this approach useful for a large number of preclinical paradigms.