Super-resolution microscopy of nuclear lamin and spindle envelope/matrix function

描述（由申请人提供）：本提案旨在使用超分辨率显微镜技术研究间期和有丝分裂期间不同类型核纤层蛋白网络/网状结构的结构组织。在目的1中，我们计划确定是否每个核纤层蛋白（LA，LC，LB 1，LB 2）形成一个单独的层网或是否建立一个层网需要在正常的间期核不同的核纤层蛋白亚型的相互作用。我们还将研究每个核纤层蛋白与染色质和核孔复合物（NPC）的相互作用。当动物细胞进入有丝分裂时，间期膜网络的部分，包括核纤层，和核蛋白重新组织成包围和渗透纺锤体微管的相互连接的网络。这种网络被称为纺锤体外壳（或鞘）或纺锤体基质。这里我们将网络称为Spindle包络/矩阵（Spenix）。定义Spenix功能的主要挑战是可视化Spenix蛋白的行为，并将其行为与有丝分裂中的功能联系起来。在目标2中，我们将应用超分辨率显微镜来研究Spenix的膜和蛋白质组织，这将打开大门，不仅解剖Spenix的组织，而且Spenix和有丝分裂纺锤体的功能关系。从拟议的研究结果将提供重要的新信息的相互作用和功能的核纤层蛋白及其相互作用与他们的主要相互作用的合作伙伴在间期和有丝分裂。

DESCRIPTION (provided by applicant): This proposal aims to study the structural organization of the different types of lamin networks/meshworks during interphase and mitosis using super resolution microscopy techniques. In Aim 1, we plan to determine whether each lamin (LA, LC, LB1, LB2) forms a separate lamina meshwork or whether the establishment of a lamina meshwork requires the interactions of different lamin isoforms in normal interphase nuclei. We will also study the interactions of each of the lamins with chromatin and nuclear pore complexes (NPCs). As animal cells enter mitosis, parts of the interphase membrane network, including the nuclear lamina, and nuclear proteins re-organize into an inter-connected network that both surrounds and permeates the spindle microtubules. This network has been referred to as either the spindle envelope (or sheath) or spindle matrix. Here we will refer to the network as the Spindle envelope/matrix (Spenix). The major challenge in defining the function of the Spenix is to visualize the behavior of Spenix proteins and link their behaviors to their functions in mitosis. In Aim 2, we will apply super resolution microscopy to study the membrane and protein organization of the Spenix, which should open the door to dissect not only the organization of the Spenix but also the functional relationship of the Spenix and the mitotic spindle. The results from the proposed studies will provide important new information regarding the interactions and functions of the nuclear lamins and their interactions with their major interacting partners in interphase and mitosis.