Retina and Optic Nerve Research Laboratory, Dalhousie University, Halifax, Nova Scotia, Canada.
Neurosci Lett. 2010 Jan 4;468(2):110-4. doi: 10.1016/j.neulet.2009.10.077. Epub 2009 Oct 29.
We investigated the specificity of cyan fluorescent protein (CFP) expression in retinal ganglion cells (RGCs) of the transgenic Thy1-CFP (B6.Cg-Tg(Thy1-CFP)23Jrs/J) mouse line, and the characteristics of these cells after optic nerve injury. RGCs of adult Thy1-CFP mice were retrogradely labeled with fluorochrome (2% fluorogold [FG]) from the superior colliculi (SC). Animals were sacrificed 7 days after RGC labeling. Retinas were fixed and whole-mounted. CFP and FG-positive cells were visualized and imaged separately. Cells positive for CFP, FG, or co-labeled were counted. In another group of animals, the left optic nerves were transected 7 days after FG labeling. They were sacrificed 7 or 21 days after transection. The retinas were whole-mounted and the characteristics of CFP-expressing cells examined. CFP-expressing cells were distributed evenly throughout the retinas of Thy1-CFP mice. The average densities of CFP and FG-positive cells in the retina were 2778+/-216 and 3230+/-157 cells/mm(2), respectively. 93.2+/-1.6% of CFP-expressing cells were also labeled with FG. However, only 79.9+/-2.5% of FG-labeled RGCs expressed CFP. The number of CFP-expressing cells decreased dramatically after transection. Cells with spindle shape, immunohistochemically identified as microglia, were seen in the retina with CFP expression at both 7 and 21 days after optic nerve transection. In retinas of Thy1-CFP mice, CFP is expressed by the large majority of RGCs, but not exclusively by RGCs. CFP is internalized by phagocytosing cells after injury to RGCs.
我们研究了转基因 Thy1-CFP（B6.Cg-Tg（Thy1-CFP）23Jrs/J）小鼠系中视网膜神经节细胞（RGC）中青色荧光蛋白（CFP）表达的特异性，以及视神经损伤后这些细胞的特征。用荧光染料（2%氟金胺[FG]）从上丘对成年 Thy1-CFP 小鼠的 RGC 进行逆行标记。在 RGC 标记后 7 天处死动物。固定并全层铺片视网膜。分别可视化和成像 CFP 和 FG 阳性细胞。计数 CFP、FG 或共标记阳性细胞。在另一组动物中，FG 标记后 7 天切断左侧视神经。它们在横切后 7 或 21 天被处死。全层铺片视网膜并检查 CFP 表达细胞的特征。CFP 表达细胞均匀分布在 Thy1-CFP 小鼠的视网膜中。视网膜中 CFP 和 FG 阳性细胞的平均密度分别为 2778+/-216 和 3230+/-157 个细胞/mm2。93.2+/-1.6%的 CFP 表达细胞也用 FG 标记。然而，只有 79.9+/-2.5%的 FG 标记的 RGC 表达 CFP。横切后 CFP 表达细胞数量急剧减少。在视神经横切后 7 和 21 天，在具有 CFP 表达的视网膜中可见到具有梭形的细胞，免疫组织化学鉴定为小胶质细胞。在 Thy1-CFP 小鼠的视网膜中，CFP 由大多数 RGC 表达，但并非仅由 RGC 表达。CFP 在 RGC 损伤后被吞噬细胞内化。
