Comparative evaluation of methods for estimating retinal ganglion cell loss in retinal sections and wholemounts.
Comparative evaluation of methods for estimating retinal ganglion cell loss in retinal sections and wholemounts.
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DOI：
10.1371/journal.pone.0110612
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Leadbeater W
中科院分区：
文献类型：
--
作者：
Mead B;Thompson A;Scheven BA;Logan A;Berry M;Leadbeater W
目的 研究对大鼠视网膜切片和视神经完整的眼睛或视神经挤压 (ONC) 后切除的眼睛的整体视网膜神经节细胞 (RGC) 进行定量的不同方法的可靠性。成年大鼠接受单侧 ONC，21 天后，在视网膜径向切片或收获前 48 小时注射 FluoroGold (FG) 的整体中对 Brn3a+、βIII-微管蛋白+ 和 Islet-1+ RGC 的数量进行定量。表型抗体标记用于区分 RGC 与星形胶质细胞、巨噬细胞/小胶质细胞和无长突细胞。在完整安装的视网膜中，具有完整视神经的眼睛中 FG+ 和 Brn3a+ RGC 的数量相似，并且在 ONC 后同样减少。当图像靠近视神经乳头拍摄时，完整组和 ONC 组之间的 RGC 数量存在较大差异。在径向切片中，Brn3a 没有对星形胶质细胞、巨噬细胞/小胶质细胞或无长突细胞染色，而 βIII-微管蛋白和 Islet-1 确实定位于无长突细胞和 RGC。在具有完整视神经的眼睛和ONC后21天的眼睛的视网膜中，βIII-微管蛋白+ RGC的数量多于Brn3a+ RGC。与 Brn3a 相比，Islet-1 染色也高估了 RGC 的数量，但仅限于 ONC 之后。与 Brn3a 染色的整体切片和 RGC 回填 FG 的视网膜整体切片相比，Brn3a 染色的径向视网膜切片中 RGC 损失的估计相似，切片具有减少实验动物使用的额外优势。
To investigate the reliability of different methods of quantifying retinal ganglion cells (RGCs) in rat retinal sections and wholemounts from eyes with either intact optic nerves or those axotomised after optic nerve crush (ONC). Adult rats received a unilateral ONC and after 21 days the numbers of Brn3a+, βIII-tubulin+ and Islet-1+ RGCs were quantified in either retinal radial sections or wholemounts in which FluoroGold (FG) was injected 48 h before harvesting. Phenotypic antibody markers were used to distinguish RGCs from astrocytes, macrophages/microglia and amacrine cells. In wholemounted retinae, counts of FG+ and Brn3a+ RGCs were of similar magnitude in eyes with intact optic nerves and were similarly reduced after ONC. Larger differences in RGC number were detected between intact and ONC groups when images were taken closer to the optic nerve head. In radial sections, Brn3a did not stain astrocytes, macrophages/microglia or amacrine cells, whereas βIII-tubulin and Islet-1 did localize to amacrine cells as well as RGCs. The numbers of βIII-tubulin+ RGCs was greater than Brn3a+ RGCs, both in retinae from eyes with intact optic nerves and eyes 21 days after ONC. Islet-1 staining also overestimated the number of RGCs compared to Brn3a, but only after ONC. Estimates of RGC loss were similar in Brn3a-stained radial retinal sections compared to both Brn3a-stained wholemounts and retinal wholemounts in which RGCs were backfilled with FG, with sections having the added advantage of reducing experimental animal usage.
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影响因子：
2.5
作者：
BUHL, EH;DANN, JF
通讯作者：
DANN, JF
影响因子：
5.3
作者：
BUNT, AH;LUND, RD
通讯作者：
LUND, RD
影响因子：
3.4
作者：
Galindo-Romero, C.;Aviles-Trigueros, M.;Agudo-Barriuso, M.
通讯作者：
Agudo-Barriuso, M.
影响因子：
56.9
作者：
BENNETT, MK;CALAKOS, N;SCHELLER, RH
通讯作者：
SCHELLER, RH
影响因子：
2
作者：
Huang, W;Fileta, J;Grosskreutz, CL
通讯作者：
Grosskreutz, CL