Neuner-Jehle M, Rhyner T A, Borbély A A
Institute of Pharmacology, University of Zürich, Switzerland.
Brain Res. 1995 Jul 10;685(1-2):143-53. doi: 10.1016/0006-8993(95)00416-n.
The mRNA level of the 17-kDa protein neurogranin (NG), a postsynaptic substrate of the protein kinase C, has previously been found to be decreased in rat forebrain after 24-h sleep deprivation (SD). To investigate the functional significance of this finding in various forebrain regions, the effect of 24-h SD on the mRNA level and the protein level of NG was determined in the cerebral cortex, hippocampus, and the total of the remaining subcortical forebrain plus midbrain areas (SFMA) of rats. In these areas, high levels of both NG mRNA and NG protein were detected by in situ hybridization and immunohistochemistry, respectively. NG protein was recognized in brain tissue by newly developed polyclonal antibodies. As determined by RNase protection assays, the level of NG mRNA was decreased in SFMA by 34 +/- 7% (P < 0.05) after 24-h SD, and was not significantly affected in the cerebral cortex and hippocampus. In contrast, on Western blots, the protein concentration of NG was reduced in the cerebral cortex by 37 +/- 7% (P < 0.05) whereas no significant changes were present in other brain areas tested. The results indicate that the mRNA and protein levels of NG are differentially modulated in rat brain by the prolongation of the waking period.
17-kDa蛋白神经颗粒素（NG）是蛋白激酶C的一种突触后底物，此前研究发现，在大鼠前脑经过24小时睡眠剥夺（SD）后，其mRNA水平会下降。为了研究这一发现对不同前脑区域的功能意义，我们测定了24小时睡眠剥夺对大鼠大脑皮层、海马以及其余皮层下前脑加中脑区域（SFMA）总和中NG的mRNA水平和蛋白水平的影响。在这些区域，分别通过原位杂交和免疫组织化学检测到高水平的NG mRNA和NG蛋白。通过新制备的多克隆抗体在脑组织中识别出NG蛋白。通过核糖核酸酶保护分析测定，24小时睡眠剥夺后，SFMA中NG mRNA水平下降了34±7%（P<0.05），而大脑皮层和海马中的NG mRNA水平未受到显著影响。相比之下，在蛋白质免疫印迹法检测中，大脑皮层中NG的蛋白质浓度降低了37±7%（P<0.05），而在其他检测的脑区中未出现显著变化。结果表明，清醒时间的延长对大鼠大脑中NG的mRNA和蛋白水平有不同的调节作用。
