Development of gene therapy by the retrograde axonal transport of adenovirus into the reeler mouse

通过将腺病毒逆行轴突转运至 reeler 小鼠中开发基因治疗

• 批准号: 11558091
• 负责人: TERASHIMA Toshio
• 金额: $ 5.25万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11558091/
• 关键词: REELER MOUSE; YOTARI MOUSE; ADENOVIRUS; RETROGRADE INFECTION; CEREBRAL CORTEX; CALLOSAL COMMISSURE; βガラクトシダーゼ; 遺伝子導入; カハールレチウス細胞; リーリン; カハール・レチウス細胞; 神経奇形マウス; 遺伝子治療

In this project, we aimed to develop the efficient introduction of exogeneous genes into the central nervous system of the neurological mutant mice, reeler and yatari. The reeler and yotari mouse are auto somal recessive, mutant mice, caused by mutation of reelin and disabled, homolog l (Dab1) gene, respectively. The mutant mouse is recognized by unstable gait and tremor and by \ I early deaths, around at the time of weaning. The cytoarchitectures of cerebeller and cerebral cortices and hippocampal formation of the reeler and yotari mouse are abnormal. In the present study, Lac-Z recombinant adenovirus was injected into the motor crtex of the normal, reeler, and yotari mice to examine how the virus is efficiently incorporated into the nervous system of the experimental animals. In the contralateral cortex, callosal commissure neurons were retrogradely labeled by the lac-Z recombinant adenovirus. The distribution pattern of CG neurons of the yotari was similar to that of the reeler : retrogradely labeled CG neurons were seen throughout all depths of the contralateral Fr1. We clarified that the small volume of the viral solution (〜0.02ul) is enough for the retrograde infection of neurons in the coerebral cortex both in the normal and reeler-phenotype mutants.

在本项目中，我们旨在开发外源基因高效导入神经突变小鼠reeler和yatari的中枢神经系统。reeler和yotari小鼠分别是由reelin和残疾的同源基因1 （Dab1）突变引起的常染色体隐性突变小鼠。突变小鼠的特征是步态不稳定和震颤，以及在断奶前后过早死亡。reeler和yotari小鼠的小脑和大脑皮层的细胞结构以及海马的形成都异常。在本研究中，将Lac-Z重组腺病毒注射到正常小鼠、reeler小鼠和yotari小鼠的运动皮层中，以研究病毒如何有效地融入实验动物的神经系统。在对侧皮层，胼胝体连接神经元被lacz重组腺病毒逆行标记。在对侧Fr1的所有深度均可见到逆行标记的CG神经元。我们证实，小体积的病毒溶液（~ 0.02ul）足以逆行感染正常和reel -表型突变体的大脑皮层神经元。

项目成果

Woodhams PL: "Aberrant trajectory of entorhino-dentate axons in the mutant Shaking Rat Kawasaki : a DiI-labelling study"European Journal of Neuroscience. 12(7). 2707-2720 (2000)

Woodhams PL：“突变体川崎震动鼠中内鼻齿状轴突的异常轨迹：一项 DiI 标记研究”《欧洲神经科学杂志》。

Okabe, S., Miwa, A., 0kado, H: "Alternative splicing of the C-terminal domain regulates cell surface expression of the NMDA receptor NR1 subunit"J neuroscience. 19(18). 7781-7792 (1999)

Okabe, S.、Miwa, A.、0kado, H：“C 末端结构域的选择性剪接调节 NMDA 受体 NR1 亚基的细胞表面表达”J 神经科学。

Sawamoto K, Nakao N, Kobayashi, K, Matsushita N, Takahashi H, Kakishita, K, Yamamoto A, Yoshizaki, T, Terashima T, Murakami F, Itakura, T, Okano H: "Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons"Proc Natl Acad Sci U

Sawamoto K，Nakao N，Kobayashi，K，Matsushita N，Takahashi H，Kakishita，K，Yamamoto A，Yoshizaki，T，Terashima T，Murakami F，Itakura，T，Okano H：“中脑的可视化、直接分离和移植

Yamashita S, Mita S, Arima T, Maeda Y, Kimura E, Nishida Y, Murakami T, Okado H, Uchino M: "Bcl-2 expression by retrograde transport of adenoviral vectors with Cre-loxP recombination system in motor neurons of mutant SOD1 transgenic mice"Gene Ther. 8(13).

Yamashita S、Mita S、Arima T、Maeda Y、Kimura E、Nishida Y、Murakami T、Okado H、Uchino M：“突变型 SOD1 运动神经元中通过 Cre-loxP 重组系统逆行转运腺病毒载体表达 Bcl-2

寺島俊雄: "組換えアデノウイルスを用いた神経回路標識法"蛋白質核酸酵素. 45(3). 537-547 (2000)

Toshio Terashima：“使用重组腺病毒的神经回路标记方法”蛋白质核酸酶45（3）（2000）。