Peng Jian-Ping, Kou Yu-Hui, Deng Jiu-Xu, Zhang Pei-Xun, Yin Xiao-Feng, Jiang Bao-Guo
Department of Orthopeadics, Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai 200092, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2013 Oct 18;45(5):807-14.
To generate peripheral nerve animal model of pure motor nerve fibers/pure sensory nerve fibers, and identify them.
The SPF SD rats were adopted in this study, and divided into 3 groups. In group A, we ablated L2-L4 ventral roots (VRs) to generate peripheral nerve animal model of pure sensory fibers. In group B, we ablated L2-L4 dorsal root ganglions (DRGs) to generate peripheral nerve animal model of pure motor fibers. Two time end-points were set as 2 weeks and 4 weeks. Neuron cells in lumbar spinal cords were detected by immunohistochemical staining with antibody of neuronal nuclei (NeuN). Motor neuron cells in lumbar spinal cords of pure motor fiber animal models and sensory neuron cells in lumbar spinal cords of pure sensory fiber animal models were counted respectively, and then compared to that of normal animals. Femoral nerves distal to the furcation were stained in osmium tetroxide, and then myelinated nerve fibers in the muscle branch and cutaneous branch of femoral nerve were counted respectively.
The mean numbers of sensory neuron cells and motor neuron cells in normal lumbar spinal cords were 62.57 ± 1.02 and 29.73 ± 3.03 per 10 × 20 visual field respectively. For different end-points, the mean numbers of sensory neuron cells after ablating vental foots were 62.12 ± 1.77 (2 weeks), 62.15 ± 1.32 (4 weeks) per 10 × 20 visual field respectively; the mean numbers of motor neuron cells after ablating DRGs were 30.12 ± 0.44 (2 weeks), 30.00 ± 1.87 (4 weeks) per 10 × 20 visual field respectively. In group A, motor axons in muscle branch were degenerated as the sensory axons in muscle branch and cutaneous branch were not changed. The senory axons in femoral nerve for the two end-points were 1 558.17 ± 50.14 (2 weeks) and 1 544.00 ± 47.42 (4 weeks). In group B, sensory axons in muscle branch were degenerated as the motor axons were reserved. The motor axons in muscle branch for the two end-points were 387.67 ± 48.50 (2 weeks) and 393.50 ± 27.86 (4 weeks). There was no statistically significant difference in these mean numbers for the two end-points. The degenerating axons and myelin sheath had not been totally eliminated by the endpoint of 2 weeks.
Peripheral nerve animal model of pure motor fibers can be generated by ablating L2-L4 DRGs; peripheral nerve animal model of pure sensory fibers can be generated by ablating L2-L4 ventral roots. The degenerating axons and myelin sheath have been totally eliminated by the end-point of 4 weeks. Ablating the ventral roots does not influence the survival of sensory neuron cells; and ablating the DRGs does not influence the survival of motor neuron cells.
构建纯运动神经纤维/纯感觉神经纤维的周围神经动物模型,并对其进行鉴定。
本研究采用SPF级SD大鼠,分为3组。A组切断L2-L4腹根以构建纯感觉纤维的周围神经动物模型。B组切断L2-L4背根神经节以构建纯运动纤维的周围神经动物模型。设定2周和4周两个时间点。用神经元细胞核(NeuN)抗体免疫组化染色检测腰段脊髓中的神经元细胞。分别计数纯运动纤维动物模型腰段脊髓中的运动神经元细胞和纯感觉纤维动物模型腰段脊髓中的感觉神经元细胞,然后与正常动物进行比较。用四氧化锇对分叉远端的股神经进行染色,然后分别计数股神经肌支和皮支中的有髓神经纤维。
正常腰段脊髓中感觉神经元细胞和运动神经元细胞的平均数分别为每10×20视野62.57±1.02个和29.73±3.03个。对于不同的时间点,切断腹根后感觉神经元细胞的平均数分别为每10×20视野62.12±1.77个(2周)、62.15±1.32个(4周);切断背根神经节后运动神经元细胞的平均数分别为每10×20视野30.12±0.44个(2周)、30.00±1.87个(4周)。A组肌支中的运动轴突发生退变,而肌支和皮支中的感觉轴突未改变。两个时间点股神经中的感觉轴突分别为1 558.17±50.14条(2周)和1 544.00±47.42条(4周)。B组肌支中的感觉轴突发生退变,而运动轴突得以保留。两个时间点肌支中的运动轴突分别为387.67±48.50条(2周)和393.50±27.86条(4周)。这两个时间点的这些平均数差异无统计学意义。到2周时间点时,退变的轴突和髓鞘尚未完全清除。
切断L2-L4背根神经节可构建纯运动纤维的周围神经动物模型;切断L2-L4腹根可构建纯感觉纤维的周围神经动物模型。到4周时间点时,退变的轴突和髓鞘已完全清除。切断腹根不影响感觉神经元细胞的存活;切断背根神经节不影响运动神经元细胞的存活。