Belliveau D J, Naus C C
Centre for Neuronal Survival, Montreal Neurological Institute, Que., Canada.
Dev Neurosci. 1995;17(2):81-96. doi: 10.1159/000111277.
We investigated the developmental expression and cellular resolution of connexin32 and 43 mRNA in the rat brain using in situ hybridization. Utilizing 35S-labelled probes, in situ hybridization was performed on sections of embryonic day 20 and postnatal days 3, 10, 15, 30 and adult brain. Connexin32 mRNA was first detected in brainstem nuclei at postnatal day 3 and in the midbrain at postnatal day 15. The level of this message continued to increase to postnatal day 30 where the level of message reached a plateau or slightly decreased by adulthood. The distribution of signal included the medial vestibular nucleus, dorsal paragigantocellular nucleus and fibre tracts of the midbrain and brainstem such as the cerebellar white matter, spinal trigeminal tract, and decussation of the superior cerebellar peduncle, areas containing predominantly neurons or oligodendrocytes. Connexin43 mRNA was first detected much earlier than connexin32 and was found in the leptomeniges of the E20 brain. It was found at all ages examined and distributed homogenously in the regions of the brain examined, suggesting its presence in astrocytes. The connexin43 riboprobe also hybridized strongly to the ependymal cells of the fourth ventricle and cerebral aqueduct. These results describe the cellular resolution of connexin mRNAs during development and that a differential pattern of expression exists in the various cell populations in the central nervous system.
我们采用原位杂交技术研究了大鼠脑中连接蛋白32和43 mRNA的发育表达及细胞定位。利用35S标记的探针，对胚胎第20天以及出生后第3、10、15、30天和成年大鼠脑的切片进行原位杂交。连接蛋白32 mRNA在出生后第3天首次在脑干核中被检测到，在出生后第15天在中脑中被检测到。该信使RNA的水平持续增加至出生后第30天，此时信使RNA水平达到平台期或在成年期略有下降。信号分布包括内侧前庭核、背侧巨细胞旁核以及中脑和脑干的纤维束，如小脑白质、脊髓三叉神经束和小脑上脚交叉，这些区域主要包含神经元或少突胶质细胞。连接蛋白43 mRNA比连接蛋白32更早被检测到，在胚胎第20天的软脑膜中被发现。在所有检测的年龄段均能发现它，且在所检测的脑区中均匀分布，提示其存在于星形胶质细胞中。连接蛋白43核糖探针也与第四脑室和中脑导水管的室管膜细胞强烈杂交。这些结果描述了连接蛋白mRNA在发育过程中的细胞定位，以及在中枢神经系统的各种细胞群体中存在差异表达模式。
