Efficiency and fidelity in mitotic spindle assembly

细胞分裂（有丝分裂）的目的是将遗传信息以染色体的形式平均分配到细胞中。 两个子细胞我们研究的首要目标是揭示使纺锤体 快速组装，错误最少。我们以前的工作表明， 可以通过几种替代路线到达主轴组件。这种替代机制的多样性 提示的假设，一个适当的平衡的贡献，从个别机制必须是 保持以确保无错误的染色体分离。目标的父1 R35 GM 130298 -01 （有丝分裂纺锤体组装的效率和保真度）是通过定量表征 正常细胞与染色体不稳定（CIN）细胞中的纺锤体组装， 染色体为了实现这一目标，我们确定并表征了控制依恋的分子机制 通过复杂的成像如激光显微手术， 染色体运动的精确跟踪，以及对染色体进行的相关电子显微镜分析。 其行为在活细胞中被跟踪直到固定的时刻的动粒。这些方法依赖于 一个激光显微照射工作站，在PI的实验室设计，组装和维护，并提供支持 来自国家卫生研究院的赠款和沃兹沃斯中心的资金。本申请旨在获得资金， 升级工作站，使激光操作能够在细胞中进行，其中各种细胞器用不同的颜色标记。 多色能力将大大加速我们的科学进步，实现父母资助的目标，并将 还促进了由NIH GMS赠款支持的几项合作研究。

The goal of cell division (mitosis) is to partition genetic information, in the form of chromosomes, equally into the two daughter cells. The overarching goal of our research is to reveal the mechanisms that allow the spindle to assemble rapidly and with minimal number of errors. Our previous work demonstrates that every major step in spindle assembly can be reached via several alternative routes. This multiplicity of alternative mechanisms prompts the hypothesis that a proper balance in the contributions from individual mechanisms must be maintained to ensure error-free chromosome segregation. The goal of the parent 1 R35 GM130298-01 (Efficiency and fidelity in mitotic spindle assembly) is to test this hypothesis by quantitatively characterizing spindle assembly in normal vs. chromosomally instable (CIN) cells that frequently mis-segregate their chromosomes. Towards this goal, we identify and characterize molecular mechanisms that govern attachment of chromosomes to microtubules of the mitotic `spindle' via sophisticated imaging such as laser microsurgery, precise tracking of chromosome movements, and correlative electron-microscopy analyses conducted on the kinetochores whose behavior was followed in live cells up to the moment of fixation. These approaches rely on a laser microirradiation workstation, designed, assembled, and maintained in the PI's laboratory with support from the NIH grants and funds contributed by the Wadsworth Center. This application seeks to obtain funds for upgrading the workstation to enable laser operations in cells with various organelles labeled in different colors. Multi-color capabilities will vastly accelerate our scientific progress towards the goals of the parent grant and will also facilitate several collaborative studies supported by NIH GMS grants.