Targetted transfection of adenovirus vector carrying brain-derived neurotrophic factoe gene prevents loss of mouse anterior horn neurons in vivo sustaining mechanical compression.

研究设计.腺病毒介导的脑源性神经营养因子（BDNF）基因转移后脑源性神经营养因子（BDNF）、胆碱乙酰转移酶（ChAT）和乙酰胆碱酯酶（AChE）在骨质增生小鼠（twy/twy）颈脊髓机械压迫区域及其周围的表达和定位的免疫组织化学分析。目的：探讨腺病毒-脑源性神经营养因子（AdV-BDNF）基因转染对自发性慢性压迫脊髓前角神经元的保护作用。一些研究报道了神经营养素对受损脊髓运动神经元的神经保护作用。然而，没有报道描述了靶向逆行神经营养基因递送对类似脊髓病病变的颈髓慢性压迫性病变中运动神经元存活的体内作用。腺病毒载体介导的LacZ标记基因（AdV-LacZ）被用于评估逆行性胰腺炎的发生。 ...更多信息 在成年twy小鼠（16周龄）和癌症研究所（ICR）小鼠（对照）中，从胸骨乳突肌到颈脊髓前角神经元的肝硬化。AdV-LacZ或AdV-BDNF注射4周后，将压迫的颈脊髓整块取出，用于□-半乳糖苷酶活性的免疫组织学研究和BDNF的免疫反应性和免疫印迹分析。采用Nissl、ChAT和ACNE染色法计数前角神经元数目。AdV-LacZ靶向肌肉注射后，成功转染twy和ICR小鼠C1和C3节段脊髓副运动神经元。AdV-BDNF-基因转染的twy小鼠对BDNF的免疫反应性明显强于AdV-LacZ-基因转染的小鼠。在显示最大压迫的脊髓水平，Adv-BDNF转染的twy小鼠的前角神经元数量在Nissl、ChAT和AChE染色的样品的拓扑神经元细胞计数中显著高于Adv-LacZ注射的twy小鼠。通过胸锁乳突肌靶向AdV-BDNF基因传递显著增加了Nissl染色的前角神经元，其缺乏核染色质溶解并显示神经突分支，并增强了Twy小鼠前角神经元中的ChAT和ACNE免疫反应性。我们的研究结果表明，有针对性的逆行腺病毒-BDNF-基因在体内传递可能会提高神经元的存活，即使在慢性机械压迫。少

Study Design. Immunohistochemical analysis of the expression and localization of brain-derived neurotrophic factor (BDNF), chorine acetyltransferase (ChAT), and activity of acetylcholine esterase (AChE) 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 anterior horn neurons.Summary of Background Data. Several studies reported the neuroprotective effects of neurotrophins on injured spinal cord motoneurons. However, no report has described the effect of targeted retrograde neurotrophic gene delivery on motoneuron survival in vivo 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 de … More livery from the stemomastoid muscle to the cervical spinal cord anterior horn neurons in adult twy mice (16-week-old) and Institute of Cancer Research (ICR) mice (control). Four weeks after the AdV-LacZ or AdV-BDNF injection, the compressed cervical spinal cord was removed en bloc for immunohistological investigation of □-galactosidase activity and immunoreactivity and immunoblot analyses of BDNF. The number of anterior horn neurons was counted using Nissl, ChAT and ACNE 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.Condusion. Targeted AdV-BDNF-gene delivery via the sternomastoid muscle significantly increased Nissl-stained anterior horn neurons, which lacked nuclear chromatolysis and showed neurite arborization, and enhanced ChAT and ACNE immunoreactivities in the twy mouse anterior horn neurons. Our results suggest that targeted retrograde AdV-BDNF-gene in vivo delivery may enhance neuronal survival even under chronic mechanical compression. Less