Dai C, Huang X, Zhao H, Wang Z, Li K
Department of Otorhinolaryngology, Eye Ear Nose and Throat Hospital, Medical Center of Fudan University, Shanghai 200031, China.
Zhonghua Er Bi Yan Hou Ke Za Zhi. 2001 Feb;36(1):10-3.
To explore the expression of bcl-2 in facial motoneurons and its subcellular distribution.
Wistar rats were used in this study. Facial nerve transection was performed at stylomastoid foramen or internal acoustic meatus. Facial nerve crush was made at stylomastiod foramen. The animal survived for 1, 3, 7, 15, 30 and 60 days respectively. Facial nucleus was treated with bcl-2 monoclonal antibody or bcl-2 DIG-labelling probe and studied with immunohistochemistry and in situ hybridization. The bcl-2 positive motoneuron was investigated with immuno-electron microscope.
It was demonstrated that bcl-2 protein level was corresponded with bcl-2 mRNA expression. The level of bcl-2 expression in facial motoneurons was high in normal facial nerve. It increased on the first day and declined on the third day post-transection in facial motoneuron. It reached the lowest level on the 15th days following facial nerve injury (P < 0.05). The expression recovered to normal level in two months (P > 0.05). After facial nerve transected, the reduction of bcl-2 expression was more significant when facial nerve transected close to facial nucleus than that far from facial nucleus (P < 0.05). Comparing to facial nerve transection in stylomastoid foramen, there was more intensive bcl-2 expression following facial nerve crush (P < 0.05). Further study showed that bcl-2 primarily resided in the nuclear envelop, endoplasmic reticulum and mitochondrial membrane.
These data indicated that high level bcl-2 protein may prevent facial motoneuron death following facial nerve injury. It is suggested that overexpression bcl-2 by transgene may prevent facial motoneurons death.
探讨bcl-2在面神经运动神经元中的表达及其亚细胞分布。
本研究采用Wistar大鼠。在茎乳孔或内耳道进行面神经横断。在茎乳孔进行面神经挤压。动物分别存活1、3、7、15、30和60天。用bcl-2单克隆抗体或bcl-2地高辛标记探针处理面神经核,并用免疫组织化学和原位杂交进行研究。用免疫电子显微镜观察bcl-2阳性运动神经元。
结果表明,bcl-2蛋白水平与bcl-2 mRNA表达一致。正常面神经中面神经运动神经元的bcl-2表达水平较高。面神经横断后第1天升高,第3天下降。面神经损伤后第15天达到最低水平(P<0.05)。两个月后表达恢复到正常水平(P>0.05)。面神经横断后,靠近面神经核横断时bcl-2表达的降低比远离面神经核时更显著(P<0.05)。与茎乳孔面神经横断相比,面神经挤压后bcl-2表达更强烈(P<0.05)。进一步研究表明,bcl-2主要位于核膜、内质网和线粒体膜。
这些数据表明,高水平的bcl-2蛋白可能预防面神经损伤后面神经运动神经元的死亡。提示通过转基因过表达bcl-2可能预防面神经运动神经元的死亡。