An in vitro model of adult mammalian nerve repair.
An in vitro model of adult mammalian nerve repair.
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DOI：
10.1016/j.expneurol.2009.05.022
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发表时间：
2010-05
影响因子：
5.3
通讯作者：
Brushart, Thomas M.
中科院分区：
文献类型：
--
作者：
Vyas, Alka;Li, Zhaobo;Aspalter, Manuela;Feiner, Jeffrey;Hoke, Ahmet;Zhou, Chunhua;O'Daly, Andres;Abdullah, Madeel;Rohde, Charles;Brushart, Thomas M.
通路衍生的生长因子在支持外周轴突再生中的作用仍然难以捉摸。很少有合适的基因敲除小鼠可用，基因沉默技术很少100%有效。为了克服这些困难，我们开发了一种体外器官型共培养系统，该系统准确地模拟了成年哺乳动物的外周神经修复。来自在其神经元中表达YFP的P4小鼠的脊髓切片用于神经支配P4外周神经的节段。这个重建的前根然后被切断并连接到神经移植物上。轴突穿过神经修复并进入移植物的生长可以用荧光显微镜重复成像以确定再生速度，并且可以以逆行方式标记亲本神经元以识别贡献神经元。从成年小鼠收获的神经移植物通过形态和功能标准在培养中保持活力。运动神经元在培养的第一周用GDNF支持，之后它们在轴突切断术中存活，因此在功能上成熟。该平台可以通过使用来自任何经遗传修饰的小鼠的运动神经元来修饰，所述小鼠可以被繁殖以表达XFP，通过从任何来源收获神经移植物，或通过用抗体、生长因子或途径抑制剂系统性地处理培养物。再生环境被控制在体内不可能的程度，并且实验动物的使用大大减少。因此，这种技术提供的灵活性和控制应该使其成为再生生物学研究的有用工具。
The role of pathway-derived growth factors in the support of peripheral axon regeneration remains elusive. Few appropriate knock-out mice are available, and gene silencing techniques are rarely 100% effective. To overcome these difficulties, we have developed an in vitro organotypic co-culture system that accurately models peripheral nerve repair in the adult mammal. Spinal cord sections from P4 mice that express YFP in their neurons are used to innervate segments of P4 peripheral nerve. This reconstructed ventral root is then transected and joined to a nerve graft. Growth of axons across the nerve repair and into the graft can be imaged repeatedly with fluorescence microscopy to define regeneration speed, and parent neurons can be labeled in retrograde fashion to identify contributing neurons. Nerve graft harvested from adult mice remains viable in culture by both morphologic and functional criteria. Motoneurons are supported with GDNF for the first week in culture, after which they survive axotomy, and are thus functionally adult. This platform can be modified by using motoneurons from any genetically modified mouse that can be bred to express XFP, by harvesting nerve graft from any source, or by treating the culture systemically with antibodies, growth factors, or pathway inhibitors. The regeneration environment is controlled to a degree not possible in vivo, and the use of experimental animals is reduced substantially. The flexibility and control offered by this technique should thus make it a useful tool for the study of regeneration biology.
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影响因子：
15.3
作者：
Ingebrigtsen, R
通讯作者：
Ingebrigtsen, R
影响因子：
4.2
作者：
Lee, YS;Baratta, J;Robertson, RT
通讯作者：
Robertson, RT
影响因子：
82.9
作者：
Kerschensteiner, M;Schwab, ME;Misgeld, T
通讯作者：
Misgeld, T
DOI：
10.1002/neu.480250702
发表时间：
1994-07-01
期刊：
JOURNAL OF NEUROBIOLOGY
影响因子：
--
作者：
LI, LX;OPPENHEIM, RW;HOUENOU, LJ
通讯作者：
HOUENOU, LJ
DOI：
10.1007/bf01611730
发表时间：
1986-08-01
期刊：
JOURNAL OF NEUROCYTOLOGY
影响因子：
--
作者：
CRANG, AJ;BLAKEMORE, WF
通讯作者：
BLAKEMORE, WF