Munemasa Yasunari, Ohtani-Kaneko Ritsuko, Kitaoka Yasushi, Kumai Toshio, Kitaoka Yuka, Hayashi Yasuhiro, Watanabe Minoru, Takeda Hiroyuki, Hirata Kazuaki, Ueno Satoki
Department of Ophthalmology, St. Marianna University School of Medicine, Kawasaki-shi, Kanagawa, Japan.
J Neurosci Res. 2006 Apr;83(5):907-18. doi: 10.1002/jnr.20786.
We examined the role of c-Jun on N-methyl-D-aspartate (NMDA)-induced neurotoxicity in the rat retina. An increase in c-Jun mRNA, c-Jun protein and phosphorylated c-Jun (p-c-Jun) levels in the retina was detected 3 hr after intravitreal injection of NMDA (20 nmol). These levels peaked after 12 hr, and then returned to their control levels by 24 hr. c-Jun and p-c-Jun immunoreactivities were observed in the retinal ganglion cell layer (RGCL), especially in retinal ganglion cells (RGCs), and in the inner nuclear layer (INL) 12 hr after NMDA injection, and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL)-positive cells were immunopositive for c-Jun and p-c-Jun. A c-Jun antisense oligodeoxynucleotide (AS ODN), which was simultaneously injected with NMDA, penetrated the cells in the RGCL and the INL, suppressed the NMDA-induced increase in c-Jun and p-c-Jun protein levels and reduced the number of TUNEL-positive cells in the RGCL 12 hr after the injection. The protective effect of c-Jun AS ODN on the NMDA-treated retina was also shown by the RGCL cell count and measurement of the IPL thickness, as well as by quantitative real-time PCR analysis of Thy-1 mRNA 7 days after the injection. These results suggest that c-Jun synthesis and phosphorylation participate in NMDA-induced neuronal cell death.
我们研究了c-Jun在N-甲基-D-天冬氨酸（NMDA）诱导的大鼠视网膜神经毒性中的作用。玻璃体内注射NMDA（20 nmol）后3小时，检测到视网膜中c-Jun mRNA、c-Jun蛋白和磷酸化c-Jun（p-c-Jun）水平升高。这些水平在12小时后达到峰值，然后在24小时恢复到对照水平。NMDA注射12小时后，在视网膜神经节细胞层（RGCL），尤其是视网膜神经节细胞（RGCs）以及内核层（INL）中观察到c-Jun和p-c-Jun免疫反应性，并且末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）阳性细胞对c-Jun和p-c-Jun呈免疫阳性。与NMDA同时注射的c-Jun反义寡脱氧核苷酸（AS ODN）穿透RGCL和INL中的细胞，抑制NMDA诱导的c-Jun和p-c-Jun蛋白水平升高，并减少注射后12小时RGCL中TUNEL阳性细胞的数量。注射7天后，通过RGCL细胞计数、IPL厚度测量以及Thy-1 mRNA的定量实时PCR分析，也显示了c-Jun AS ODN对NMDA处理的视网膜的保护作用。这些结果表明，c-Jun的合成和磷酸化参与了NMDA诱导的神经元细胞死亡。
