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

描述(申请人提供)：这项建议旨在利用超分辨率显微镜技术研究不同类型的板层网络/网状结构在间期和有丝分裂期间的结构组织。在目标1中，我们计划确定每个层粘连蛋白(LA、Lc、Lb1、Lb2)是否形成一个单独的层网络，或者一个层网络的建立是否需要正常间期核中不同的层粘连蛋白亚型的相互作用。我们还将研究每个层与染色质和核孔复合体(NPC)的相互作用。随着动物细胞进入有丝分裂，包括核膜在内的间期膜网络的部分部分和核蛋白重新组织成一个相互连接的网络，该网络包围并渗透着纺锤体微管。这种网络被称为主轴包络(或鞘)或主轴矩阵。在这里，我们将网络称为主轴包络/矩阵(Spenix)。定义Spenix功能的主要挑战是可视化Spenix蛋白的行为，并将它们的行为与它们在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研究Spenix的膜和蛋白质组织，这将打开解剖Spenix的组织以及Spenix和有丝分裂纺锤体的功能关系的大门。这些研究的结果将为核层的相互作用和功能，以及它们在间期和有丝分裂中与主要相互作用伙伴的相互作用提供重要的新信息。

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.