Yoshida K, Imaki J, Fujisawa H, Harada T, Ohki K, Matsuda H, Hagiwara M
Department of Ophthalmology, Hokkaido University School of Medicine, Sapporo, Japan.
Invest Ophthalmol Vis Sci. 1996 Jan;37(1):174-9.
To examine the expression of c-fos proto-oncogene and phosphorylation of cAMP responsive element binding (CREB) protein in the rat retina after changes in the light-dark condition.
Rats were exposed to both steady light and flashing light and were killed at the end of light exposure. The retinas were analyzed by in situ hybridization using single-stranded RNA probes for c-fos transcripts and by immunocytochemistry using phosphoSer-133 specific CREB antiserum, anti-calcium calmodulin dependent protein (CaM) kinase II, and anti-CaM kinase IV.
c-fos mRNA was expressed in the outer half of the inner nuclear layer (INL) and in the ganglion cell layer (GCL) after 30 minutes of sustained light. After 30 minutes of flashing light, c-fos expression also was detected in the inner border of the INL. Phosphorylated CREB immunoreactive nuclei had similar distribution after steady and flashing light. Both CaM kinase II and CaM kinase IV, which phosphorylate CREB at Ser 133 in vitro, were expressed in the GCL and in the INL. CaM kinase II, however, was localized in the inner border of the INL, whereas CaM kinase IV was distributed in the outer half of the INL.
These results suggest that the differential expression of c-fos mRNA induced by flashing and sustained light may reflect the CREB phosphorylation by CaM kinases in a different subpopulation of retinal cells.
研究明暗条件改变后大鼠视网膜中c-fos原癌基因的表达及环磷酸腺苷反应元件结合蛋白(CREB)的磷酸化情况。
将大鼠暴露于持续光和闪烁光下,在光照结束时处死。通过使用针对c-fos转录本的单链RNA探针进行原位杂交以及使用磷酸化丝氨酸133特异性CREB抗血清、抗钙调蛋白依赖性蛋白(CaM)激酶II和抗CaM激酶IV进行免疫细胞化学分析视网膜。
持续光照30分钟后,c-fos mRNA在内核层(INL)外半部和神经节细胞层(GCL)表达。闪烁光照30分钟后,在INL内边界也检测到c-fos表达。持续光和闪烁光后,磷酸化CREB免疫反应性细胞核分布相似。在体外可使CREB在丝氨酸133位点磷酸化的CaM激酶II和CaM激酶IV均在GCL和INL表达。然而,CaM激酶II定位于INL内边界,而CaM激酶IV分布于INL外半部。
这些结果表明,闪烁光和持续光诱导的c-fos mRNA差异表达可能反映了视网膜细胞不同亚群中CaM激酶对CREB的磷酸化作用。