The distribution and time course of brain-derived neurotrophic factor expression after plasmid DNA transfer to adult rat spinal cord

质粒DNA转移至成年大鼠脊髓后脑源性神经营养因子表达的分布和时程

• 批准号: 13671499
• 负责人: UCHIDA Kenzo
• 金额: $ 2.24万
• 依托单位: FUKUI MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671499/
• 关键词: gene transfer; liposome; brain-derived neurotrophic factor; cDNA; spinal cord injury; in vivo; rat

We investigated the potential of non-viral vector-mediated brain-derived neurotrophic factor (BDNF) gene transfection in adult rat spinal cord. The potential of transfection of in vivo spinal cord was examined using three cationic lipids, Lipofectin^<TM>, LipofectAMINE^<TM> and LipofectAMINE^<TM> 2000, which have been proven to transfect genes into neuronal cells in vitro using plasmid DNA. After the direct injection of liposome cDNA complexes into thoracic spinal cord, histological analysis and immunoblot analysis demonstrated green fluorescent protein (GFP) and exogenous BDNF expression only in the group of LipofectAMINE^<TM> 2000/ cDNA complexes. In the group of LipofectAMINE^<TM> 2000/ cDNA complexes, fluorescent histochemical examination confirmed the presence of injected GFP around the injection areas for at least 5 days after injection. The transfected cells were identified reactive astrocytes, and some neuron. Both BDNF mRNA using RT-PCR and BDNF proteins using Western Blotting indicated a similar profile. The results suggest that cationic liposome-mediated delivery may be a practical method for gene transfer in spinal cord.

我们研究了非病毒载体介导的脑源性神经营养因子（BDNF）基因转染成年大鼠脊髓的潜力。使用三种阳离子脂质Lipofectin^<TM>、LipofectAMINE^<TM>和LipofectAMINE^<TM>2000检查体内脊髓转染的潜力，已经证明使用质粒DNA在体外将基因转染到神经元细胞中。在将脂质体cDNA复合物直接注射到胸脊髓中后，组织学分析和免疫印迹分析表明仅在LipofectAMINE^ 2000/ cDNA复合物组中表达绿色荧光蛋白（GFP）和外源性BDNF<TM>。在LipofectAMINE^<TM>2000/ cDNA复合物组中，荧光组织化学检查证实注射后至少5天注射区域周围存在注射的GFP。经鉴定，转染细胞为反应性星形胶质细胞和部分神经元。RT-PCR检测的BDNF mRNA和Western Blotting检测的BDNF蛋白均显示相似的表达谱。结果表明，阳离子脂质体介导的基因转移可能是一种实用的脊髓基因转移方法。

项目成果

Yamaura, I., Yone, K., Nakahara, S., Nagamine, T., Baba, H., Uchida, K., Komiya, S.: "Mechanism of destructive pathologic changes in the spinal cord under chronic mechanical compression"Spine. 27. 21-26 (2002)

Yamaura, I.、Yone, K.、Nakahara, S.、Nagamine, T.、Baba, H.、Uchida, K.、Komiya, S.：“慢性机械压迫下脊髓破坏性病理变化的机制”

Uchida, K., Baba, H., Maezawa, Y., Kubota, C.: "Progressive changes of neurofilament proteins and growth-associated protein-43 immunoreactivities at the site of cervical spinal cord compression in spinal hyperostotic mice"Spine. 27. 480-486 (2002)

Uchida, K.、Baba, H.、Maezawa, Y.、Kubota, C.：“脊柱骨质增生小鼠颈髓受压部位神经丝蛋白和生长相关蛋白 43 免疫反应性的进行性变化”。

Uchida, K., Baba, H., Furukawa, S., Omiya, M., Kokubo, Y., Kubota, C., Nakajima.: "Increased expression of neurotrophins and their receptors in the mechanically compressed spinal cord of the spinal hyperostotic mouse (twy/twy)"Acta neuropathologica. (2003

Uchida, K.、Baba, H.、Furukawa, S.、Omiya, M.、Kokubo, Y.、Kubota, C.、Nakajima.：“脊髓机械压缩的脊髓中神经营养因子及其受体的表达增加

Uchida, K., Baba, H., Furukawa, S., Omiya, M., Kokubo, Y., Kubota, C., Nakajima.: "Increased expression of neurotrophins and their receptors in the mechanically compressed spinal cord of the spinal hyperostotic mouse (twy/twy)"Acta neuropathologica. 106-1

Uchida, K.、Baba, H.、Furukawa, S.、Omiya, M.、Kokubo, Y.、Kubota, C.、Nakajima.：“脊髓机械压缩的脊髓中神经营养因子及其受体的表达增加

Yamaura, I., Yone, K., Nakahara, S., Nagamine, T Baba, H., Uchida, K., Komiya, S: "Mechanism of destructive pathologic changes in the spinal cord under chronic mechanical compression"Spine. 27. 21-26 (2002)

Yamaura, I.、Yone, K.、Nakahara, S.、Nagamine, T Baba, H.、Uchida, K.、Komiya, S：“慢性机械压迫下脊髓破坏性病理变化的机制”脊柱。