Cytological study in plant morphogenesis.

植物形态发生的细胞学研究。

基本信息

批准号：
13640661
负责人：
HASEZAWA Seiichiro
金额：
$ 2.24万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13640661/
关键词：
microtubule cell cycle GFP cell synchronization tobacco BY-2 cells vacuole TVM Atvam3p BY-GT16細胞 FM4-64

项目摘要

We have observed CMT reorganization by immunofluorescence microscopy, and reported that MTs were nucleated from daughter nuclear surfaces prior to the CMTs (Kumagai and Hasezawa, 2001). However, we could not precisely define the time-course of events because of the rapid transition from phragmoplasts to CMTs. To observe such rapidly occurring events, we used the green fluorescent protein (GFP) and α-tubulin fusion protein as probes of MTs in living cells (Kumagai et al., 2001). In addition, we attempted to use the dye, FM4-64, to visualize vacuoles in living cells (Kutsuna and Hasezawa, 2002). This dye, first taken up into the lipid bilayer of plasma membranes and then finally incorporated into vacuolar membranes via the endocytotic pathway, unexpectedly enables us to observe dynamic vacuolar movements during cell cycle transition. We also confirmed the dynamic changes of VM structures by the observation using transgenic BY-2 cells expressing GFP-AtVam3p fusion protein (BY-GV). Furthermore, by using transgenic BY-2 cells that stably express a GFP-tubulin fusion protein (BY-GT16, Kumagai et al. 2001), we could study the relationship between the dynamics of vacuoles and microtubules.By combining these methods, we are therefore able to discuss the mechanisms by which daughter cells prepare the machinery for cell expansion in the proper directions. It is known that after their formation, CMTs are stabilized into the direction appropriate for the cell type, geometry, and circumstance. In this study, we also concentrate on the dynamic behavior of MTs during M/G_1 transition, and leave the details of how CMTs, once formed, are stabilized into proper directions.

我们通过免疫荧光显微镜观察到 CMT 重组，并报道 MT 在 CMT 之前从子核表面成核（Kumagai 和 Hasezawa，2001）。然而，由于从子核表面的快速转变，我们无法精确定义事件的时间进程。为了观察这种快速发生的事件，我们使用绿色荧光蛋白（GFP）和α-微管蛋白融合蛋白作为 MT 的探针。活细胞中的液泡（Kumagai 等，2001）此外，我们尝试使用染料 FM4-64 来可视化活细胞中的液泡（Kutsuna 和 Hasezawa，2002）。质膜，然后最终通过内吞途径融入液泡膜，出人意料地使我们能够观察细胞周期转变过程中的动态液泡运动。通过使用表达GFP-AtVam3p融合蛋白的转基因BY-2细胞（BY-GV）观察VM结构的动态变化此外，通过使用稳定表达GFP-微管蛋白融合蛋白的转基因BY-2细胞（BY-GT16，熊谷）。 et al. 2001），我们可以研究液泡和微管动力学之间的关系。因此，通过结合这些方法，我们能够讨论子细胞制备液泡和微管的机制。众所周知，CMT 在形成后会稳定在适合细胞类型、几何形状和环境的方向。在这项研究中，我们还关注 M/期间的动态行为。 G_1 转变，并留下 CMT 一旦形成如何稳定到正确方向的细节。