ANALYSIS OF MECHANISMS OF NEURONAL CELL DEATH USING ADENOVIRUS VECTOR SYSTEM

利用腺病毒载体系统分析神经细胞死亡机制

• 批准号: 08680808
• 负责人: YAGINUMA Hiroyuki
• 金额: $ 1.6万
• 依托单位: FUKUSHIMA MEDICAL UNIVERSITY (1997)University of Tsukuba (1996)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08680808/
• 关键词: Bcl-2; Crm A; Adenovirus; chick; motoneuron; Dorsal root ganglion; apoptosis; programmed cell death; アデノウイルス; ウイルスベクタ; 神経細胞死; Bcl-2; ニワトリ; アポトーシス; 遺伝子発現; 遺伝子導入; 細胞死; ニワトリ胚; Cre-LoxP; 発生

Aims of this project were (I) to establish adenovirus vector system that is capable of introducing genes into neuronal cells of the chick embryo, and (II) to analyze effects of Bcl-2 and Crm A on neuronal cell death in vitro and in vivo. Major results are the following :(1) In the first trial, we failed to obtain recombinant adenovirus that express Bcl-2 under regulation of CAG promoter. Employing Cre-LoxP system that can regulate expression of genes, we succeeded to obtain the recombinant adenovirus that express Bcl-2.(2) It was confirmed that genes introduced by adenovirus vecotrs were expressed in the chick DRG neurons and chick motoneurons in vitro. Introduced human Bcl-2 could rescue DRG neurons and motoneurons from cell death in vitro.(3) It is also confirmed that adenovirus vectors injected into leg bud were retrogradely transported to the motoneurons in the lateral motor column. Bcl-2 introduced with this retrograde transport could rescue some motoneurons after limb bud removal.(4) In the rat C6 glioma cell line system, human Bcl-2 introduced by adenovirus vector induced cell death.(5) We could not find rescuing effects of Crm A gene introduced by adenovirus vector in the DRG neuron system.

该项目的目的是（I）建立能够将基因导入鸡胚神经元细胞的腺病毒载体系统，以及（II）分析Bcl-2和Crm A对体外和体内神经元细胞死亡的影响。主要结果如下：(1)在第一次试验中，我们未能获得在CAG启动子调控下表达Bcl-2的重组腺病毒。利用能够调控基因表达的Cre-LoxP系统，我们成功获得了表达Bcl-2的重组腺病毒。(2)体外证实腺病毒载体导入的基因在鸡DRG神经元和鸡运动神经元中表达。引入人Bcl-2可在体外挽救DRG神经元和运动神经元免于细胞死亡。(3)还证实注射入腿芽的腺病毒载体逆行转运至外侧运动柱的运动神经元。通过这种逆行转运引入的Bcl-2可以在肢芽去除后挽救一些运动神经元。(4)在大鼠C6胶质瘤细胞系系统中，腺病毒载体引入的人Bcl-2诱导细胞死亡。(5)我们没有发现腺病毒载体引入的Crm A基因对DRG神经元系统有拯救作用。

项目成果

Nobuko Shiraiwa & Hiroyuki Yaginuma: "An increase in caspase-3-like activity during early motoneuron death in the cervical cord of the chick embryo." Acta Anatomica Nipponica. 73. 357 (1998)

白岩信子

Tokoro, Y., Sugawara, T. Yaginuma, H., Nakauchi, H., Terhorst, C., Wang, B. and Takahama, Y.: "A mouse carrying genetic defect in the choice between T and B lymphocytes"J. Immunol.. 161. 4591-4598 (1998)

Tokoro, Y.、Sugara, T. Yaginuma, H.、Nakauchi, H.、Terhorst, C.、Wang, B. 和 Takahama, Y.：“在 T 和 B 淋巴细胞选择中携带遗传缺陷的小鼠”J

Yaginuma, H., M.Tomita, N.Takashita, S.E.McKay, C.Cardwell, Q.-W.Yin, and R.W.Oppenheim: "A novel type of programmed neuronal death in the cervical spinal cord of the chick embryo" J.Neurosci. 16. 3685-3703 (1996)

Yaginuma, H.、M.Tomita、N.Takashita、S.E.McKay、C.Cardwell、Q.-W.Yin 和 R.W.Oppenheim：“鸡胚颈脊髓中一种新型的程序性神经元死亡”J

H.Yaginuma et al: "A Novel type of programmed neuronal death in the ceroical spinal cord of the chick embryo" Journal of Neuroscience. 16. 3685-3703 (1996)

H.Yaginuma 等人：“鸡胚颈脊髓中一种新型的程序性神经元死亡”《神经科学杂志》。

HIROYUKI YAGINUMA, 他: "Bcl-2 Introduced with Retrograde Transport of Adenovirus Vectors Rescues Motoneurons from Cell Death in Vivo." Neuroscience Res.Suppl. 22. S330 (1998)

HIROYUKI YAGINUMA 等人：“通过腺病毒载体逆行转运引入 Bcl-2，可挽救体内细胞死亡的运动神经元。” Neuroscience Res.Suppl. 22. S330 (1998)