Gene expression and apoptosis in the spinal cord neurons after sciatic nerve injury
Gene expression and apoptosis in the spinal cord neurons after sciatic nerve injury
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DOI：
10.1016/s0197-0186(96)00077-0
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发表时间：
1997-04-01
影响因子：
4.2
通讯作者：
Liu, PK
中科院分区：
文献类型：
--
作者：
Gu, ZZ;Pan, YC;Liu, PK
为了证明脊髓运动神经元对其与靶点通讯的依赖性，我们在单侧坐骨神经断裂的Sprague-Dawley大鼠（大于或等于9周龄的雄性）中检测了腰椎（L3-L6）脊髓神经元中立即早期基因c-Sos和神经营养因子基因的表达。采用原位杂交技术，在周围神经横断45 min ~ 3 h内（每个时间点n = 4）检测脊髓节段运动神经元中c-fos mRNA的表达。免疫组化结果显示，c-fos mRNA表达与Fos抗原表达呈正相关，calbindin和parvalbumin抗原表达无明显变化。坐骨神经断裂后90 min， BDNF mRNA表达增加。然而，未观察到NGF mRNA表达增强。用末端转移酶将地高辛- dutp掺入脊髓同侧段DNA片段的3'-OH末端，观察神经损伤48 h后神经元DNA的断裂。在对照动物的脊髓中未观察到出现DNA断裂的细胞核。最后，我们观察到大多数星形胶质细胞没有DNA断裂。由于该方法检测DNA片段是早期检测细胞凋亡或程序性细胞死亡的一种方法，提示我们可以检测脊髓早期细胞死亡，并提示坐骨神经横断后脊髓内神经元的靶标依赖性。1997爱思唯尔科学有限公司
To demonstrate a dependence of spinal cord motoneurons on the communication with their targets, the expression of immediate early gene c-Sos and neurotrophin genes in the lumbar (L3-L6) spinal cord neurons was examined in Sprague-Dawley rats (male greater than or equal to 9-weeks-old) with unilateral sciatic nerve transection. Using in situ hybridization, we detected the expression of c-fos mRNA in the motoneurons of the spinal cord segments within 45 min to 3 h of peripheral nerve transection (n = 4 in each time point). The expression of c-fos mRNA was also correlated positively with the expression of Fos antigen using immunohistochemistry, while no change in calbindin and parvalbumin antigens were noted. The expression of BDNF mRNA increased al 90 min after sciatic nerve transection. However, no detectable enhancement in the expression of NGF mRNA was observed. DNA fragmentation in neurons was observed using the incorporation of digoxigenin-dUTP by terminal transferase into 3'-OH terminals of DNA fragments in the ipsilateral section of the spinal cords 48 h after nerve injury. Nuclei that exhibited DNA fragmentation were not observed in the spinal cord of the control animals. Lastly, we observed that the majority of astrocytes did not have DNA fragmentation. Because the detection of DNA fragmentation using this assay is one of early detections of apoptosis or programmed cell death, the result suggested we could detect early cell death in spinal cord, and indicated a target dependence of the neurons in the spinal cord after transection of sciatic nerve. (C) 1997 Elsevier Science Ltd.