Chesselet M F, Weiss L, Wuenschell C, Tobin A J, Affolter H U
J Comp Neurol. 1987 Aug 1;262(1):125-40. doi: 10.1002/cne.902620110.
Neurotransmitter-related messenger RNAs were detected by in situ hybridization in sections of rat and mouse brains by using 35S-radiolabelled RNA probes transcribed from cDNAs cloned in SP6 promoter-containing vectors. The distribution of messenger RNAs for glutamic acid decarboxylase, tachykinins (substance P and K), and tyrosine hydroxylase was examined in the striatum, pallidum, and substantia nigra. Dense clusters of silver grains were observed with the RNA probe complementary of the cellular messenger RNA for glutamic acid decarboxylase (antisense RNA) over most large neurons in the substantia nigra pars reticulata and medium-sized to large neurons in all pallidal subdivisions. A few very densely and numerous lightly labelled medium-sized neurons were present in the striatum. Among the areas examined, only the striatum contained neurons labelled with the antisense tachykinin RNA. Most of these neurons were of medium size, and a few were large. With the antisense tyrosine hydroxylase RNA, silver grains were found over neurons of the substantia nigra pars compacta and adjacent A10 and A8 dopaminergic cell groups. No signal was observed with RNAs identical to the cellular messenger RNA for glutamic acid decarboxylase or tachykinin (sense RNA). These results show a good correlation with immunohistochemical studies, suggesting that documented differences in the distribution and the level of glutamic acid decarboxylase, tyrosine hydroxylase, and substance P immunoreactivities in neurons of the basal ganglia are related to differences in the level of expression of the corresponding genes rather than to translation accessibility, stability, or transport of the gene products.
通过使用从克隆于含SP6启动子载体的cDNA转录而来的35S放射性标记RNA探针,在大鼠和小鼠脑切片中通过原位杂交检测神经递质相关信使RNA。在纹状体、苍白球和黑质中检测了谷氨酸脱羧酶、速激肽(P物质和神经激肽K)和酪氨酸羟化酶信使RNA的分布。在黑质网状部的大多数大神经元以及所有苍白球亚区的中大型神经元上,观察到与谷氨酸脱羧酶细胞信使RNA互补的RNA探针(反义RNA)形成的密集银粒簇。纹状体中有一些非常密集且数量众多的轻度标记的中型神经元。在所检查的区域中,只有纹状体含有用反义速激肽RNA标记的神经元。这些神经元大多数为中型,少数为大型。在用反义酪氨酸羟化酶RNA时,在黑质致密部以及相邻的A10和A8多巴胺能细胞群的神经元上发现了银粒。与谷氨酸脱羧酶或速激肽细胞信使RNA相同的RNA(正义RNA)未观察到信号。这些结果与免疫组织化学研究显示出良好的相关性,表明在基底神经节神经元中记录到的谷氨酸脱羧酶、酪氨酸羟化酶和P物质免疫反应性在分布和水平上的差异与相应基因表达水平的差异有关,而不是与基因产物的翻译可及性、稳定性或转运有关。