Targeted retrograde transfection of adenovirus vector carrying brain-derived neurotrophic factor gene prevents loss of mouse (twy/twy) anterior horn neurons in vivo sustaining mechanical compression
Targeted retrograde transfection of adenovirus vector carrying brain-derived neurotrophic factor gene prevents loss of mouse (twy/twy) anterior horn neurons in vivo sustaining mechanical compression
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DOI：
10.1097/01.brs.0000228772.53598.cc
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发表时间：
2006-08-01
期刊：
影响因子：
3
通讯作者：
Baba, Hisatoshi
中科院分区：
文献类型：
--
作者：
Xu, Kan;Uchida, Kenzo;Baba, Hisatoshi
研究设计。目的对腺病毒(Adv)介导的BDNF基因在骨质增生症小鼠颈脊髓机械性压迫区及周围的免疫组织化学分析。目的：探讨靶向AdV-BDNF基因导入对自发性慢性压迫脊髓运动神经元的神经保护作用。一些研究报道了神经营养因子对损伤脊髓的神经保护作用。然而，在类似脊髓病病变的颈髓慢性压迫性病变中，定向逆行神经营养基因治疗对运动神经元存活的影响尚未见报道。应用LacZ标记基因重组腺病毒载体(ADV-LacZ)对16周龄成年小鼠和对照组胸锁乳突肌逆行递送进行了研究。注射AdV-LacZ或AdV-BDNF 4周后，整块摘除受压的颈髓，进行b-半乳糖苷酶活性、BDNF免疫反应和免疫印迹分析。尼氏染色、ChAT染色和AChE染色计数前角神经元数量。在靶向肌肉注射后，AdV-LacZ成功地在TWY和ICR小鼠体内导入了C1节和C3节之间的脊髓副运动神经元。转导ADV-BDNF-基因的TWY小鼠对BDNF的免疫反应性明显高于转导ADV-LacZ基因的小鼠。在脊髓受压最大处，Nissl、ChAT和AChE染色标本的前角神经元数量显著高于注射ADV-LacZ的TWY小鼠。靶向注射Adv-BDNF-基因可显著增加THY内Nissl染色的前角神经元，增强胆碱能酶的活性。我们的结果表明，体内靶向逆行携带AdV-BDNF-基因可以提高神经元的存活率，即使在慢性机械压迫下也是如此。
Study Design. Immunohistochemical analysis after adenovirus (AdV)-mediated BDNF gene transfer in and around the area of mechanical compression in the cervical spinal cord of the hyperostotic mouse (twy/twy).Objective. To investigate the neuroprotective effect of targeted AdV-BDNF gene transfection in the twy mouse with spontaneous chronic compression of the spinal cord motoneurons.Summary of Background Data. Several studies reported the neuroprotective effects of neurotrophins on injured spinal cord. However, no report has described the effect of targeted retrograde neurotrophic gene delivery on motoneuron survival in chronic compression lesions of the cervical spinal cord resembling lesions of myelopathy.Methods. LacZ marker gene using adenoviral vector (AdV-LacZ) was used to evaluate retrograde delivery from the sternomastoid muscle in adult twy mice (16-week-old) and ( control). Four weeks after the AdV-LacZ or AdV-BDNF injection, the compressed cervical spinal cord was removed en bloc for immunohistologic investigation of b-galactosidase activity and immunoreactivity and immunoblot analyses of BDNF. The number of anterior horn neurons was counted using Nissl, ChAT and AChE staining.Results. Spinal accessory motoneurons between C1 and C3 segments were successfully transfected by AdV-LacZ in both twy and ICR mice after targeted intramuscular injection. Immunoreactivity to BDNF was significantly stronger in AdV-BDNF-gene transfected twy mice than in AdV-LacZ-gene transfected mice. At the cord level showing the maximum compression in AdV-BDNF-transfected twy mice, the number of anterior horn neurons was significantly higher in the topographic neuronal cell counting of Nissl-, ChAT-, and AChE-stained samples than in AdV-LacZ-injected twy mice.Conclusion. Targeted AdV-BDNF-gene delivery significantly increased Nissl- stained anterior horn neurons and enhanced cholinergic enzyme activities in the twy. Our results suggest that targeted retrograde AdV-BDNF-gene in vivo delivery may enhance neuronal survival even under chronic mechanical compression.