Mechanics of Microtubule Aster Growth and Positioning: Instrumentation Supplement

微管 Aster 生长和定位的力学：仪器补充

• 批准号: 10798982
• 负责人: JESSE C GATLIN
• 金额: $ 18.52万
• 依托单位: UNIVERSITY OF WYOMING
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-04 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10798982
• 关键词: Adopted; Biomechanics; Cell Cycle; Cell Nucleus; Cell division; Cells; Chromosome Segregation; Cytoplasm; Cytoskeleton; Development; Engineering; Ensure; Exposure to; Fertilization; Fiber; Filament; Growth; Hydrogels; Interphase; Light; Location; Mechanics; Microtubules; Mitosis; Mitotic spindle; Molecular; Organ; Pathway interactions; Pharmaceutical Preparations; Polymers; Positioning Attribute; Process; Research; Shapes; Surface; Therapeutic; Visualization; Work; instrumentation; light microscopy; male; novel strategies; parent grant; response; scaffold; sperm cell

PROJECT SUMMARY/ABSTRACT (from parent grant GM135568) The microtubule cytoskeleton is critical for organization the insides of cells. At times during the cell cycle, this network adopts the form of an astral array with microtubule fibers emanating from a central focus and branching outward throughout the cell. Importantly, the center of this array is typically found near the geometric center of the cell where it is anchored to the surface of the nucleus. Forces generated by the microtubules themselves move the internal organs of the cell during interphase, and because the network also serves as a scaffold upon which these organs attach, its position dictates their spatial arrangement within the cell. During mitosis similar forces positioning the cell division machinery and establish the eventual location of the cell division plane. Though aster centering is likely critical in all cells, it is particularly relevant in large cells immediately after fertilization. Here, the male pronucleus (from the sperm) must transverse large distances to reach the cell center and establish the location of mitotic spindle formation and division plane positioning during the subsequent mitosis. Thus, errors in this process can lead to erroneous cell division and have deleterious effects on development. Precisely how the microtubule aster generates and responds to forces to move to the cell center remains unanswered. In this work, we describe the application and continued development of a new approach engineered to overcome existing limitations inherent to other approaches used to study aster positioning. This approach relies on the use of photo-labile hydrogels, which can be polymerized and degraded in response to exposure to specific wavelengths of light. When combined with cell-free cytoplasmic extracts, it provides exquisite control of cytoplasmic shape and volume in a platform that is amenable to visualization using standard wide-field and confocal light microscopy.

项目摘要/摘要（来自家长补助金 GM135568） 微管细胞骨架对于细胞内部的组织至关重要。期间有时 细胞周期，该网络采用星体阵列的形式，微管纤维从 一个中心焦点并在整个细胞中向外分支。重要的是，这个数组的中心是 通常发现在细胞的几何中心附近，它固定在细胞的表面 核。微管本身产生的力移动细胞的内部器官 在间期期间，并且因为网络还充当这些器官的支架 附着，它的位置决定了它们在细胞内的空间排列。有丝分裂期间类似的力 定位细胞分裂机器并确定细胞分裂的最终位置 飞机。 尽管紫苑居中可能对所有细胞都至关重要，但它在大型细胞中尤其重要 受精后立即。在这里，雄性原核（来自精子）必须横穿大 到达细胞中心的距离并确定有丝分裂纺锤体形成的位置和 随后的有丝分裂期间的分割平面定位。因此，此过程中的错误可能会导致 错误的细胞分裂并对发育产生有害影响。究竟是如何 微管紫苑产生并响应力移动到细胞中心遗骸 无人回答。在这项工作中，我们描述了一种新的应用和持续开发 旨在克服其他研究方法固有的局限性的方法 紫苑定位。这种方法依赖于光不稳定水凝胶的使用，该水凝胶可以 暴露于特定波长的光下会发生聚合和降解。什么时候 与无细胞细胞质提取物相结合，可对细胞质形状进行精确控制 和体积在一个平台上，可以使用标准的宽视场和共焦进行可视化 光学显微镜。