Axotomy-induced retinal ganglion cell death in adult mice: Quantitative and topographic time course analyses
Axotomy-induced retinal ganglion cell death in adult mice: Quantitative and topographic time course analyses
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DOI：
10.1016/j.exer.2011.02.008
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发表时间：
2011-05-01
影响因子：
3.4
通讯作者：
Agudo-Barriuso, M.
中科院分区：
文献类型：
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作者：
Galindo-Romero, C.;Aviles-Trigueros, M.;Agudo-Barriuso, M.
大鼠视神经损伤后视网膜神经节细胞的命运已被彻底描述，但在小鼠中没有，尽管该物种被充分用作研究影响视网膜神经节细胞群的不同实验范式的模型。本文对眶内神经切断引起的小鼠视网膜神经节细胞丢失的过程进行了定量和局部解剖学分析。为了做到这一点，我们已经双重确定了视网膜神经节细胞在所有视网膜追踪他们的主要retinorecipient区，上级丘，并通过他们的BRN 3A（产品Pou 4f 1基因）的表达。在大鼠中，这种转录因子由大多数视网膜神经节细胞表达;然而在小鼠中，尚不知道这些神经元的整个群体中有多少表达它。因此，在这项工作中，我们也评估了BRN 3A阳性视网膜神经节细胞的总群体。使用新开发的常规方法在所有整装视网膜中自动定量这些。在对照视网膜中，使用先前报道的方法（Salinas-Navarro等人，2009年b）。然而，在视神经损伤后，追踪的视网膜神经节细胞必须通过视网膜取样手动定量，然后推断它们的总数量。在未处理的整体标本中，示踪视网膜神经节细胞的平均（+/-标准差）总数为40，437（+/- 3196），BRN 3A阳性细胞的平均（+/- 1821）总数为34，697（+/- 1821）。对于两种标志物，视网膜神经节细胞损失在轴突切开术后5天首次显著，并且到第21天，分析的最后时间点，分别只有15%或12%的示踪或BRN 3A阳性视网膜神经节细胞存活。等密度图显示，在对照视网膜中，BRN 3A和示踪视网膜神经节细胞的分布相似，沿着鼻颞轴密集在背侧视网膜。轴突切断后，BRN 3A阳性视网膜神经节细胞的进行性损失是弥漫性的，并影响整个视网膜。总之，这是第一个研究评估的价值，在总数和密度方面，视网膜神经节细胞存活轴突切断后2至21天的病变。此外，我们已经证明，BRN 3A由总视网膜神经节细胞群体的85.6%表达，并且由于BRN 3A阳性视网膜神经节细胞显示出与追踪的视网膜神经节细胞相同的空间分布和变性的时间过程，因此BRN 3A是鉴定、定量和评估该物种中离体视网膜神经节细胞损失的可靠标志物。(c)2011爱思唯尔有限公司保留所有权利。
The fate of retinal ganglion cells after optic nerve injury has been thoroughly described in rat, but not in mice, despite the fact that this species is amply used as a model to study different experimental paradigms that affect retinal ganglion cell population. Here we have analyzed, quantitatively and topographically, the course of mice retinal ganglion cells loss induced by intraorbital nerve transection. To do this, we have doubly identified retinal ganglion cells in all retinas by tracing them from their main retinorecipient area, the superior colliculi, and by their expression of BRN3A (product of Pou4f1 gene). In rat, this transcription factor is expressed by a majority of retinal ganglion cells; however in mice it is not known how many out of the whole population of these neurons express it. Thus, in this work we have assessed, as well, the total population of BRN3A positive retinal ganglion cells. These were automatically quantified in all whole-mounted retinas using a newly developed routine. In control retinas, traced-retinal ganglion cells were automatically quantified, using the previously reported method (Salinas-Navarro et al., 2009b). After optic nerve injury, though, traced-retinal ganglion cells had to be manually quantified by retinal sampling and their total population was afterwards inferred. In naive whole-mounts, the mean (+/- standard deviation) total number of traced-retinal ganglion cells was 40,437 (+/- 3196) and of BRN3A positive ones was 34,697(+/- 1821). Retinal ganglion cell loss was first significant for both markers 5 days post-axotomy and by day 21, the last time point analyzed, only 15% or 12% of traced or BRN3A positive retinal ganglion cells respectively, survived. Isodensity maps showed that, in control retinas, BRN3A and traced-retinal ganglion cells were distributed similarly, being densest in the dorsal retina along the naso-temporal axis. After axotomy the progressive loss of BRN3A positive retinal ganglion cells was diffuse and affected the entire retina. In conclusion, this is the first study assessing the values, in terms of total number and density, of the retinal ganglion cells surviving axotomy from 2 till 21 days post-lesion. Besides, we have demonstrated that BRN3A is expressed by 85.6% of the total retinal ganglion cell population, and because BRN3A positive retinal ganglion cells show the same spatial distribution and temporal course of degeneration than traced ones, BRN3A is a reliable marker to identify, quantify and assess, ex-vivo, retinal ganglion cell loss in this species. (c) 2011 Elsevier Ltd. All rights reserved.