Cytological study in plant morphogenesis.

植物形态发生的细胞学研究。

• 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

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.

我们已经通过免疫荧光显微镜观察到CMT重组，并报道了MT在CMT之前从子核表面成核（Kumagai和Hasezawa，2001）。然而，我们不能精确地定义的时间过程中的事件，因为从成膜细胞到CMT的快速过渡。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活细胞中MT的探针（Kumagai et al.，2001年）。此外，我们尝试使用染料FM 4 -64来可视化活细胞中的空泡（Kutsuna和Hasezawa，2002）。这种染料，首先进入质膜的脂质双层，然后通过内吞途径最终纳入液泡膜，出乎意料地使我们能够观察到细胞周期转换过程中的动态液泡运动。我们还通过使用表达GFP-AtVam 3 p融合蛋白的转基因BY-2细胞（BY-GV）的观察证实了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-GT 16，Kumagai et al.2001），我们可以研究空泡和微管的动力学之间的关系，因此，通过结合这些方法，我们可以讨论子细胞准备向正确方向进行细胞扩增的机制。已知在它们形成之后，CMT被稳定成适合于细胞类型、几何形状和环境的方向。在本研究中，我们也集中在MT的M/G_1转变的动力学行为，并留下详细的CMTs，一旦形成，是如何稳定到适当的方向。

项目成果

熊谷 史: "γ-Tubulin distribution during cortical microtubule reor ganization at the M/G_1 interface in tobacco BY-2 cells"European Journal of Cell Biology. 82. 43-51 (2003)

Fumi Kumagai：“烟草 BY-2 细胞 M/G_1 界面处皮质微管重组过程中的 γ-微管蛋白分布”欧洲细胞生物学杂志 82. 43-51 (2003)。

熊谷 史: "Dynamic organization of microtubules and microfilaments during cell cycle progression in higher plant cells"Plant Biology. 3. 4-16 (2001)

Fumi Kumagai：“高等植物细胞细胞周期进程中微管和微丝的动态组织”植物生物学。 3. 4-16 (2001)

Ueda K. et al.: "Development and disintegration of phragmoplasts and postmitotic MTs in living cultured cells of a GFP-TUA6 transgenic Arabidopsis plant"Protoplasma. in press. (2003)

Ueda K.等人：“GFP-TUA6转基因拟南芥植物的活培养细胞中成膜体和有丝分裂后MT的发育和分解”原生质体。

馳澤 盛一郎: "Dynamic changes and roles of cytoskeleton during cell cycle in higher plant cells"International Review of Cytology. 214. 161-191 (2002)

Seiichiro Hasezawa：“高等植物细胞细胞周期中细胞骨架的动态变化和作用”国际细胞学评论 214. 161-191 (2002)。

植田 勝巳: "Development and disintegration of phragmoplasts and postmitotic MTs in living cultured cells of a GFP-TUA6 transgenic Arabidopsis plant"Protoplasma. (in press). (2003)

Katsumi Ueda：“GFP-TUA6 转基因拟南芥植物活培养细胞中成膜体和有丝分裂后 MT 的发育和分解”原生质体（2003 年）。