Dkhissi O, Chanut E, Wasowicz M, Savoldelli M, Nguyen-Legros J, Minvielle F, Versaux-Botteri C
Laboratoire de NeuroCytologie Oculaire, Institut National de la Santé et de la Recherche Médicale (INSERM), Paris, France.
Invest Ophthalmol Vis Sci. 1999 Apr;40(5):990-5.
To investigate whether retinal cell death observed in an avian glaucoma-like disorder occurs by apoptosis and whether an increase in excitotoxic amino acid concentration in the vitreous humor is associated temporally with cell death in the retina.
Presumptive retinal apoptotic nuclei were identified by histochemical detection of DNA fragmentation (by TdT-dUTP terminal nick-end labeling [TUNEL]), and vitreal concentrations of glutamate and several other amino acids were determined by high-pressure liquid chromatography with fluorometric detection in the al mutant quail (Coturnix coturnix japonica) in which a glaucoma-like disorder develops spontaneously.
TUNEL-labeled nuclei were located mostly in the ganglion cell layer (GCL) in the retina of mutant quails 3 months after hatching. However, labeled nuclei were also observed in the inner and outer nuclear layers. At 7 months, most TUNEL-positive nuclei were detected in the inner nuclear layer, whereas labeled cells in the GCL were reduced in number. No TUNEL-labeled nuclei were detected in the retina of control quails at any age. Vitreal concentrations of glutamate and aspartate were significantly increased in 1-month-old mutant quails compared with control animals. Concentrations decreased at 3 months, and no significant differences were observed between strains at 7 months.
Presumptive apoptotic cell death is detected from 3 months after hatching in mutant quails and is not restricted to retinal ganglion cells. Cell death appears just after a significant increase in excitotoxic amino acid concentrations in the vitreous humor, suggesting a correlation between both events.
研究在一种禽类青光眼样疾病中观察到的视网膜细胞死亡是否通过凋亡发生,以及玻璃体液中兴奋性毒性氨基酸浓度的增加是否与视网膜中的细胞死亡在时间上相关。
通过DNA片段化的组织化学检测(通过TdT-dUTP末端脱氧核苷酸转移酶介导的缺口末端标记法[TUNEL])来鉴定推测的视网膜凋亡细胞核,并通过高压液相色谱结合荧光检测法测定al突变鹌鹑(日本鹌鹑)玻璃体液中谷氨酸和其他几种氨基酸的浓度,该突变鹌鹑会自发发展出青光眼样疾病。
孵化后3个月的突变鹌鹑视网膜中,TUNEL标记的细胞核大多位于神经节细胞层(GCL)。然而,在内核层和外核层也观察到了标记的细胞核。在7个月时,大多数TUNEL阳性细胞核在内核层被检测到,而GCL中的标记细胞数量减少。在任何年龄的对照鹌鹑视网膜中均未检测到TUNEL标记的细胞核。与对照动物相比,1月龄突变鹌鹑玻璃体液中谷氨酸和天冬氨酸的浓度显著升高。3个月时浓度下降,7个月时两品系之间未观察到显著差异。
在突变鹌鹑孵化后3个月即可检测到推测的凋亡细胞死亡,且不仅限于视网膜神经节细胞。细胞死亡恰好在玻璃体液中兴奋性毒性氨基酸浓度显著增加之后出现,提示这两个事件之间存在相关性。