权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Analysis of early infection with poliovirus in motor neurons.

运动神经元脊髓灰质炎病毒早期感染分析。

基本信息

批准号：
21790438
负责人：
OHKA Seii
金额：
$ 2.75万
依托单位：
独立行政法人国立がん研究センター (2010)The University of Tokyo (2009)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2009
资助国家：
日本
起止时间：
2009 至 2010
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21790438/
关键词：
ポリオウイルス運動神経細胞

项目摘要

We were able to culture primary motor neurons from rat or mouse using dual chamber system. It enabled us to culture the synapse side and cell body side in the different medium. To improve the culture condition, we improved the condition of the medium. The axonal elongation has improved by adding the extract of skeletal muscles from mouse. We cultured motor neurons derived from human poliovirus (PV) receptor (hPVR) - transgenic (Tg) mouse on the dual chamber and added PV defective interfering particles coding GFP from the synapse side or the cell body side. When the virus was added from the cell body side, expression of GFP was observed at six hours after the addition. On the other hand, when the virus was added from the synapse side, expression of GFP had not been observed until twenty-four hours after the addition, while the expression was observed at forty-eight hours after the addition. These results suggest that the infection from the cell body side progresses differently to the infection from the synapse side. Next, we tried to optimize the four-dimensional imaging system for live fluorescent-cells. For long term culture under the microscope, we changed the medium into CO2 independent medium in a closed system. This enabled cells to live longer under the microscope. We tried various conditions of laser intensity, pinhole size, and number of Z slices for long term observation under the confocal laser microscope. However, we could not optimize the conditions for the long term observation as long as the confocal laser microscope was used.

我们用双室系统培养了大鼠或小鼠的原代运动神经元。它使我们能够在不同的培养基中培养突触侧和细胞体侧。为了改善培养条件，我们改进了培养基条件。添加小鼠骨骼肌提取物可提高小鼠轴突伸长。我们在双腔室培养来自人脊髓灰质炎病毒（PV）受体（hPVR）转基因（Tg）小鼠的运动神经元，并在突触侧或细胞体侧添加编码GFP的PV缺陷干扰颗粒。从细胞体侧加入病毒，加入后6小时观察GFP的表达。另一方面，当从突触侧添加病毒时，直到添加后24小时才观察到GFP的表达，而在添加后48小时才观察到表达。这些结果表明，来自细胞体侧的感染与来自突触侧的感染进展不同。接下来，我们尝试优化活荧光细胞的四维成像系统。为了在显微镜下长期培养，我们将培养基改为封闭系统中不依赖CO2的培养基。这使得细胞在显微镜下存活的时间更长。在激光共聚焦显微镜下，我们尝试了激光强度、针孔大小、Z片数量等多种条件进行长期观察。然而，只要使用激光共聚焦显微镜，我们就无法优化长期观察的条件。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

ホームページ等。

主页等

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Analysis of dissemination pathways for poliovirus.

脊髓灰质炎病毒传播途径分析。

DOI：
发表时间：
2009
期刊：
ウイルス 59(1)
影响因子：
0
作者：
Goto M.;Shimada K.;Sato A.;Takahashi E.;Fukasawa T.;Takahashi T.;Ohka S.;Taniguchi T.;Honda E.;Nomoto A.;Ogura A.;Kirikae T.;Hanaki K.;Ohka S.
通讯作者：
Ohka S.

運動神経細胞におけるポリオウイルス感染初期過程の解析

脊髓灰质炎病毒感染运动神经元的初始过程分析