Laser-based single-axon transection for high-content axon injury and regeneration studies.
Laser-based single-axon transection for high-content axon injury and regeneration studies.
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基于激光的单轴横断，用于高内核轴突损伤和再生研究。
DOI：
10.1371/journal.pone.0026832
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Costantino S
中科院分区：
文献类型：
--
作者：
Kunik D;Dion C;Ozaki T;Levin LA;Costantino S
研究神经元对轴突损伤的再生反应对于开发新的轴突保护疗法至关重要。在这里，我们研究激光横断后的视网膜神经元 RGC-5 细胞系，证明这些细胞启动再生反应的能力与轴突长度和损伤后细胞运动相关。我们证明，低能量皮秒激光脉冲可以在体外实现未标记的单轴突的横切，并以微米精度精确诱导损伤。我们建立了实现轴突横断的条件，并使用延时显微镜表征了 RGC-5 轴突再生和细胞体反应。我们开发了一种算法来分析细胞轨迹，并建立了损伤后细胞运动、轴突长度和再生反应启动之间的相关性。轴突切除的 RGC-5 细胞运动反应的特征表明，能够修复或再生受损轴突的细胞比不能修复或再生的细胞迁移得更慢。此外，我们还发现，具有长轴突的 RGC-5 细胞无法恢复受损的轴突，而且此类细胞的运动能力更强。我们描述的平台可以通过亚细胞分辨率和细胞培养物中高内涵筛选的性能来高度控制轴突损伤。
The investigation of the regenerative response of the neurons to axonal injury is essential to the development of new axoprotective therapies. Here we study the retinal neuronal RGC-5 cell line after laser transection, demonstrating that the ability of these cells to initiate a regenerative response correlates with axon length and cell motility after injury. We show that low energy picosecond laser pulses can achieve transection of unlabeled single axons in vitro and precisely induce damage with micron precision. We established the conditions to achieve axon transection, and characterized RGC-5 axon regeneration and cell body response using time-lapse microscopy. We developed an algorithm to analyze cell trajectories and established correlations between cell motility after injury, axon length, and the initiation of the regeneration response. The characterization of the motile response of axotomized RGC-5 cells showed that cells that were capable of repair or regrowth of damaged axons migrated more slowly than cells that could not. Moreover, we established that RGC-5 cells with long axons could not recover their injured axons, and such cells were much more motile. The platform we describe allows highly controlled axonal damage with subcellular resolution and the performance of high-content screening in cell cultures.
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影响因子：
3.4
作者：
Lieven, Christopher J.;Millet, Lucia E.;Levin, Leonard A.
通讯作者：
Levin, Leonard A.
DOI：
10.3389/fnene.2010.00021
发表时间：
2010-01-01
期刊：
Frontiers in neuroenergetics
影响因子：
--
作者：
Allegra Mascaro, Anna Letizia;Sacconi, Leonardo;Pavone, Francesco S
通讯作者：
Pavone, Francesco S
影响因子：
3.4
作者：
Botvinick, EL;Venugopalan, V;Berns, MW
通讯作者：
Berns, MW
影响因子：
4.4
作者：
Frassetto, LJ;Schlieve, CR;Levin, LA
通讯作者：
Levin, LA
DOI：
10.1016/s0169-328x(00)00224-2
发表时间：
2001-01-31
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
--
作者：
Krishnamoorthy, RR;Agarwal, P;Agarwal, N
通讯作者：
Agarwal, N