Kiyama H., Emson P. C., Ruth J.
MRC Group, AFRC Institute of Animal Physiology & Genetics Research, Babraham, Cambridge CB2 4AT, UK.
Eur J Neurosci. 1990;2(6):512-524. doi: 10.1111/j.1460-9568.1990.tb00442.x.
The expression of tyrosine hydroxylase mRNA in the rat brain was examined using a novel alkaline phosphatase labelled antisense oligodeoxynucleotide probe. The alkaline phosphatase labelled probe revealed the presence of tyrosine hydroxylase mRNA in all the major cell groups and cell bodies previously described as containing catecholamine fluorescence or known to contain tyrosine hydroxylase immunoreactivity. Using standardized development protocols qualitative comparisons between the amount of mRNA signal in different adrenergic, noradrenergic or dopaminergic cell groups could be made. These studies showed that of the three known catecholaminergic cell types the level of tyrosine hydroxylase mRNA signal was high in the noradrenergic and dopaminergic cells, but much lower in the adrenergic cell groups. The sensitivity of this nonradioactive method of in situ hybridization is excellent and has considerable potential for studies of coexistence or coexpression of two mRNA signals for the localization of mRNA signals at the electron-microscope level.
使用一种新型碱性磷酸酶标记的反义寡脱氧核苷酸探针检测大鼠脑中酪氨酸羟化酶mRNA的表达。碱性磷酸酶标记的探针显示,在先前描述的所有主要细胞群和细胞体中均存在酪氨酸羟化酶mRNA,这些细胞群和细胞体含有儿茶酚胺荧光或已知含有酪氨酸羟化酶免疫反应性。采用标准化的显影方案,可以对不同肾上腺素能、去甲肾上腺素能或多巴胺能细胞群中的mRNA信号量进行定性比较。这些研究表明,在三种已知的儿茶酚胺能细胞类型中,酪氨酸羟化酶mRNA信号水平在去甲肾上腺素能和多巴胺能细胞中较高,但在肾上腺素能细胞群中则低得多。这种非放射性原位杂交方法的灵敏度极佳,在研究两个mRNA信号的共存或共表达以及在电子显微镜水平定位mRNA信号方面具有相当大的潜力。