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）建立能够将基因引入雏鸡胚胎的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。主要结果是：（1）在第一次试验中，我们未能获得在CAG启动子调节下表达Bcl-2的重组腺病毒。使用可以调节基因表达的CRE-LoxP系统，我们成功地获得了表达Bcl-2的重组腺病毒。（2）证实，腺病毒vecotrs引入的基因在Chick DRG神经元和体外的Chick Motoneurons中表达。引入的人Bcl-2可以在体外从细胞死亡中拯救DRG神经元和运动神经元。（3）也证实，注入腿部芽中的腺病毒载体被逆行转移到侧运动柱中的运动神经元。用这种逆行运输引入的BCL-2可以在去除肢体芽后挽救一些运动神经元。（4）在大鼠C6胶质瘤细胞系系统中，由腺病毒载体诱导的细胞死亡引入的人类Bcl-2。（5）我们找不到CRM的促进adnovirus vector在DRG神经元系统中引入的基因。

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

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)

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

Y. Sukawara T.、Yaginuma, H. 等人：“在 T 和 B 淋巴细胞选择中携带遗传缺陷的小鼠。”J.Immunol.161（1998 年）。