Mawatari Yuki, Fukushima Mikiko, Inoue Toshihiro, Setoguchi Takao, Taga Tetsuya, Tanihara Hidenobu
Department of Ophthalmology and Visual Science, Kumamoto University Graduate School of Medical Sciences, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
Brain Res. 2005 Sep 7;1055(1-2):7-14. doi: 10.1016/j.brainres.2005.06.047.
The purpose of this study was to investigate the differentiation of neural progenitor cells (NPCs) following retinal transplantation in N-methyl-D-aspartate (NMDA)-treated eyes. NMDA was injected into the vitreous cavity of adult rat eyes. NPCs were prepared from telencephalic neuroepithelium of enhanced green fluorescence protein (EGFP) transgenic mice on embryonic day 14.5. A cell suspension was injected into the vitreous cavity in experimental eyes. Immunohistochemistry was conducted at 1, 2 or 4 weeks after transplantation of NPCs in an effort to determine the survival and differentiation of transplanted NPCs. Similar experiments were conducted using glycoprotein (gp)130-null (-/-) mice. Examination of retinal sections revealed that transplanted NPCs could survive for at least 4 weeks in NMDA-treated retinas. Immunohistochemical studies for specific cell-type markers revealed that, among the transplanted NPCs at 2 weeks after transplantation, the mean percentage (+/-standard deviation) of glial fibrillary acidic protein (GFAP)-positive (glial) cells was 63.5 +/- 7.4%, demonstrating the differentiation of transplanted NPCs with a preference for the glial lineage. Furthermore, the mean percentage of betaIII-tubulin-positive (mature neuronal) cells was 18.8 +/- 4.5%. Following transplantation of NPCs isolated from gp130-/- mice into NMDA-treated retinas, the mean percentage of GFAP-positive cells (17.6 +/- 7.0%), was significantly lower than that in NPCs isolated from wild-type mice (59.1 +/- 6.0%, P = 0.04, Mann-Whitney U test). Preferential differentiation of NPCs into the glial lineage is induced through gp130 signaling in NMDA-treated eyes.
本研究的目的是调查在经N-甲基-D-天冬氨酸（NMDA）处理的眼睛中进行视网膜移植后神经祖细胞（NPC）的分化情况。将NMDA注入成年大鼠眼睛的玻璃体腔。NPC取自胚胎第14.5天增强型绿色荧光蛋白（EGFP）转基因小鼠的端脑神经上皮。将细胞悬液注入实验眼的玻璃体腔。在移植NPC后1、2或4周进行免疫组织化学，以确定移植NPC的存活和分化情况。使用糖蛋白（gp）130基因敲除（-/-）小鼠进行了类似实验。视网膜切片检查显示，移植的NPC在经NMDA处理的视网膜中可存活至少4周。针对特定细胞类型标志物的免疫组织化学研究显示，移植后2周的移植NPC中，胶质纤维酸性蛋白（GFAP）阳性（胶质）细胞的平均百分比（±标准差）为63.5±7.4%，表明移植的NPC优先向胶质细胞谱系分化。此外，βIII-微管蛋白阳性（成熟神经元）细胞的平均百分比为18.8±4.5%。将从gp130-/-小鼠分离的NPC移植到经NMDA处理的视网膜后，GFAP阳性细胞的平均百分比（17.6±7.0%）显著低于从野生型小鼠分离的NPC（59.1±6.0%，P = 0.04，曼-惠特尼U检验）。在经NMDA处理的眼睛中，通过gp130信号传导诱导NPC优先向胶质细胞谱系分化。
