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Studies on Electrofusim of Plant Protoplasts Using High Speed Microscopy

植物原生质体高速显微镜电纺研究

基本信息

批准号：
07640530
负责人：
TANIGUCHI Mieko
金额：
$ 1.47万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07640530/
关键词：
Protoplast High speed microscopy Actin Filament Electrofusion Network パルス電場蛍光染色植物

项目摘要

Actin is an important component of plant cytoskeleton that not only involved in contributing to the cytoarchitecture of cells but also in cell motility. Cell fusion is an important process for studying the development of the cell and for the subsequent cell division. The study is to report on three dimensional structures and dynamic aspects of plant protoplasts during early process of electrofusion. 1) New technology for recording the time-resolved phenomena has developed using a streak camera and a polarization microscope. We obtained that the technique is very useful in the study of muscle contraction and cell fusion. 2) We obtained sub microsecond and microsecond rotational motions with in the contractile myofibrils by analyzing the time-resolved birefringence. 3) We analyzed the location and movement of actin filaments after the application with AC field and DC pulse. 4) We observed that bundles of actin filaments are forming cables and networks 5) Three dimensional observations of protoplasts were also made by a high voltage electron microscope (1 MeV) equipped a revolving stage to get a pair of stereo-micrographs. Ring structure of actin filaments appeared at the contact areas between fused cells. We found the joint particle connecting between actin filaments. 6) A soft X-ray microscope with zone plate was installed at UVSOR.The X-ray micrographs of dry and wet protophasts were taken. X-ray micrographs of protoplasts were compared with those of optical and electron micrographs.

肌动蛋白是植物细胞骨架的重要组成部分，不仅参与细胞的细胞结构，而且参与细胞运动。细胞融合是研究细胞发育和随后细胞分裂的重要过程。该研究旨在报告电融合早期过程中植物原生质体的三维结构和动态方面。 1）使用条纹相机和偏振显微镜记录时间分辨现象的新技术已经开发出来。我们发现该技术在肌肉收缩和细胞融合的研究中非常有用。 2）通过分析时间分辨双折射，我们获得了收缩肌原纤维的亚微秒和微秒旋转运动。 3）我们分析了施加交流场和直流脉冲后肌动蛋白丝的位置和运动。 4）我们观察到肌动蛋白丝束正在形成电缆和网络。 5）还通过配备旋转台的高压电子显微镜（1MeV）对原生质体进行了三维观察，以获得一对立体显微照片。肌动蛋白丝的环形结构出现在融合细胞之间的接触区域。我们发现肌动蛋白丝之间有连接颗粒。 6)在UVSOR处安装带波带板的软X射线显微镜。拍摄干和湿原菌的X射线显微照片。将原生质体的 X 射线显微照片与光学和电子显微照片进行比较。