Blits Bas, Carlstedt Thomas P, Ruitenberg Marc Jan, de Winter Fred, Hermens Wim T J M C, Dijkhuizen Paul A, Claasens Jill W C, Eggers Ruben, van der Sluis Ronald, Tenenbaum Liliane, Boer Gerard J, Verhaagen Joost
Graduate School Neurosciences Amsterdam, Netherlands Institute for Brain Research, 1105 AZ Amsterdam, The Netherlands.
Exp Neurol. 2004 Oct;189(2):303-16. doi: 10.1016/j.expneurol.2004.05.014.
Following avulsion of a spinal ventral root, motoneurons that project through the avulsed root are axotomized. Avulsion between, for example, L2 and L6 leads to denervation of hind limb muscles. Reimplantation of an avulsed root directed to the motoneuron pool resulted in re-ingrowth of some motor axons. However, most motoneurons display retrograde atrophy and subsequently die. Two neurotrophic factors, glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF), promote the survival of motoneurons after injury. The long-term delivery of these neurotrophic factors to the motoneurons in the ventral horn of the spinal cord is problematic. One strategy to improve the outcome of the neurosurgical reinsertion of the ventral root following avulsion would involve gene transfer with adeno-associated viral (AAV) vectors encoding these neurotrophic factors near the denervated motoneuron pool. Here, we show that AAV-mediated overexpression of GDNF and BDNF in the spinal cord persisted for at least 16 weeks. At both 1 and 4 months post-lesion AAV-BDNF- and -GDNF-treated animals showed an increased survival of motoneurons, the effect being more prominent at 1 month. AAV vector-mediated overexpression of neurotrophins also promoted the formation of a network of motoneuron fibers in the ventral horn at the avulsed side, but motoneurons failed to extent axons into the reinserted L4 root towards the sciatic nerve nor to improve functional recovery of the hind limbs. This suggests that high levels of neurotrophic factors in the ventral horn promote sprouting, but prevent directional growth of axons of a higher number of surviving motoneurons into the implanted root.
脊髓腹侧神经根撕脱后,通过撕脱神经根投射的运动神经元会发生轴突切断。例如,L2和L6之间的撕脱会导致后肢肌肉失神经支配。将撕脱的神经根重新植入到运动神经元池可导致一些运动轴突重新生长。然而,大多数运动神经元会出现逆行性萎缩并随后死亡。两种神经营养因子,即胶质细胞源性神经营养因子(GDNF)和脑源性神经营养因子(BDNF),可促进损伤后运动神经元的存活。将这些神经营养因子长期递送至脊髓腹角的运动神经元存在问题。一种改善神经根撕脱后神经外科重新植入效果的策略是,使用编码这些神经营养因子的腺相关病毒(AAV)载体在失神经的运动神经元池附近进行基因转移。在此,我们表明,AAV介导的脊髓中GDNF和BDNF的过表达持续了至少16周。在损伤后1个月和4个月时,接受AAV-BDNF和-GDNF治疗的动物均显示运动神经元存活率增加,这种效应在1个月时更为显著。AAV载体介导的神经营养因子过表达还促进了撕脱侧腹角运动神经元纤维网络的形成,但运动神经元未能将轴突延伸至重新植入的L4神经根并朝向坐骨神经,也未能改善后肢的功能恢复。这表明腹角中高水平的神经营养因子促进了发芽,但阻止了更多存活运动神经元的轴突向植入的神经根定向生长。