A study on PPB-associated molecules by the HP freezing.

通过 HP 冷冻对 PPB 相关分子进行研究。

• 批准号: 08044208
• 负责人: MINEYUKI Yoshinobu
• 金额: $ 1.22万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08044208/
• 关键词: High Pressure Freezing; Electron Microscope; Microtubules; Preprophase band; Root tips of Allium; Cell division; Cytokinesis; Division Plane Orientation; プレプロフェースバンド

To know the mechanism of division plane oriention is an important problem for plant morphogenesis. Just befor cells enter the mitosis, Microtubules (Mts) gather to form a preprophase band (PPB). As the PPB locates at the site where cell plate will be fused at the end of cytokinesis, to know how Mts are organized as a PPB is essential for understanding regulatory nmechanisms of the division plane orientation. Actins and cdks are reported to be in PPBs but their role have not yet clear. In oreder to know the relationship among these molecules and Mts in a PPB,electron microscopic (EM) study is necessary. However, we could not get reliable results by the conventional EM because Mts are very dynamic structure. In order to solve this problem, we need to introduce high pressure freeezing (HPF) method. In the present study, we examine onion root tips by HPF to know whether HPF is useful for our purpose. Results showed that we could get better PPB images of the thin section of Spurr's resin by HPF than by the chemical fixation. In order to examine the possibility of the three dimentional analysis at the EM level, freeze fracture and deep teching mehtods are applied to the HPF samples. As our approach seems fracture and deep teching mehtods are applied to the HPF samples. As our approach seems to be promissing, we concluded that this project is worth to continue in future.

了解分面方向的机制是植物形态建成的一个重要问题。就在细胞进入有丝分裂之前，微管 (Mts) 聚集形成前期带 (PPB)。由于 PPB 位于细胞板在胞质分裂结束时融合的位点，了解 Mts 如何组织为 PPB 对于理解分裂面方向的调控机制至关重要。据报道，肌动蛋白和 cdks 存在于 PPB 中，但它们的作用尚不清楚。为了了解 PPB 中这些分子与 Mts 之间的关系，需要进行电子显微镜 (EM) 研究。然而，由于 Mts 是非常动态的结构，因此我们无法通过传统的 EM 获得可靠的结果。为了解决这个问题，我们需要引入高压冷冻（HPF）方法。在本研究中，我们通过 HPF 检查洋葱根尖，以了解 HPF 是否对我们的目的有用。结果表明，与化学固定相比，通过 HPF 可以获得更好的 Spurr 树脂薄片的 PPB 图像。为了检验电磁水平三维分析的可能性，对 HPF 样品应用了冷冻断裂和深加工方法。由于我们的方法似乎将断裂和深度技术方法应用于 HPF 样品。由于我们的方法似乎很有希望，我们得出的结论是该项目值得在未来继续下去。