Yoshimura T, Satake M, Kobayashi T
Department of Neurology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
J Neurochem. 1996 Sep;67(3):1252-8. doi: 10.1046/j.1471-4159.1996.67031252.x.
That many cells express more than one connexin (Cx) led us to examine whether Cxs other than Cx32 are expressed in the PNS. In addition to Cx32 mRNA, Cx43 and Cx26 mRNAs were detected in rat sciatic nerve by northern blot analysis. Cx43 mRNA, but not Cx26 mRNA, was expressed in both the primary Schwann cell culture and immortalized Schwann cell line (T93). The steady-state levels of the Cx43 mRNA in the primary Schwann cell culture increased 2.0-fold with 100 microM forskolin, whereas that of Po increased 7.0-fold. Immunoreactivity to Cx43 was detected on western blots of cultured Schwann cells, T93 cells, and sciatic nerves but not on blots of PNS myelin. Immunohistochemical study using human peripheral nerves revealed that anti-Cx43 antibody stained cytoplasm around nucleus of Schwann cells but not myelin, confirming western blot results. Although Po expression was markedly decreased by crush injury of the sciatic nerves, Cx43 expression showed no apparent change. Developmental profiles showed that Cx43 expression in the sciatic nerve increased rapidly after birth, peaked at about postnatal day 6, and then decreased gradually to a low level. In adult rats, the Cx43 mRNA value was much lower than that of Cx32. These findings suggest that Cx43 is localized in Schwann cell bodies and that, compared with Po, its expression is less influenced by axonal contact and cyclic AMP levels. The high expression on postnatal day 6 indicates that Cx43 may be related to PNS myelination. Cx43 is another gap junction, but its function appears to differ from that of Cx32, as judged by the differences in their localization and developmental profiles.
许多细胞表达不止一种连接蛋白(Cx),这促使我们研究除Cx32之外的其他Cx是否在周围神经系统(PNS)中表达。通过Northern印迹分析,在大鼠坐骨神经中除检测到Cx32 mRNA外,还检测到了Cx43和Cx26 mRNA。Cx43 mRNA而非Cx26 mRNA在原代雪旺细胞培养物和永生化雪旺细胞系(T93)中均有表达。原代雪旺细胞培养物中Cx43 mRNA的稳态水平在100 microM福司可林作用下增加了2.0倍,而Po的稳态水平增加了7.0倍。在培养的雪旺细胞、T93细胞和坐骨神经的蛋白质印迹上检测到了对Cx43的免疫反应性,但在PNS髓磷脂的印迹上未检测到。使用人周围神经的免疫组织化学研究表明,抗Cx43抗体染色雪旺细胞核周围的细胞质,但不染色髓磷脂,证实了蛋白质印迹结果。虽然坐骨神经挤压伤后Po表达明显降低,但Cx43表达没有明显变化。发育情况显示,坐骨神经中Cx43表达在出生后迅速增加,在出生后第6天左右达到峰值,然后逐渐下降至低水平。在成年大鼠中,Cx43 mRNA值远低于Cx32。这些发现表明Cx43定位于雪旺细胞体,并且与Po相比,其表达受轴突接触和环磷酸腺苷水平的影响较小。出生后第6天的高表达表明Cx43可能与PNS髓鞘形成有关。Cx43是另一种间隙连接,但从其定位和发育情况的差异判断,其功能似乎与Cx32不同。