3D STRUCTURE OF ER IN MITOTIC AND INTERPHASE CELLS

有丝分裂和间期细胞中 ER 的 3D 结构

• 批准号: 7955061
• 负责人: MARK S LADINSKY
• 金额: $ 1.07万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7955061
• 关键词: Anaphase; Area; Binding; Cell Nucleus; Cell division; Cells; Cellular Structures; Cercopithecus pygerythrus; Complement; Computer Retrieval of Information on Scientific Projects Database; Cytoplasm; Data Set; Endoplasmic Reticulum; Freezing; Funding; Grant; Immunoelectron Microscopy; Immunofluorescence Immunologic; Institution; Interphase; Interphase Cell; Kidney; Light; Metaphase; Mitosis; Mitotic; Mitotic spindle; Molecular; Nuclear Envelope; Pattern; Phase; Prometaphase; Prophase; Research; Research Personnel; Resources; Ribosomes; Source; Staging; Structure; Surface; United States National Institutes of Health; Work; electron tomography; fluorescence microscope; telophase; three dimensional structure

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are using electron tomography of rapidly-frozen, freeze-substituted African green monkey kidney (BSC1) cells to evaluate changes to the fine structure of the endoplasmic reticulum (ER) during the different stages of cell division. Three dimensional fluorescence microscope studies of whole mitotic cells suggest that the ER network undergoes morphological changes in each phase of mitosis, all of which differ from the interphase ER pattern. Electron tomography is being used to complement the LM work and extend the findings to the fine structural level. We have so far prepared 9 tomographic datasets of interphase cells, 19 sets of cells in metaphase, 3 sets of anaphase and 5 sets of telophase cells. Initial results confirm the presence of cisternal and reticular ER domains in interphase BSC1 cells. Although these domains are continuous, they are largely situated in different parts of the cytoplasm; Cisternal domains are nearer the nucleus while the reticular areas are nearer the cell periphery. During mitosis, most notably in metaphase and anaphase, ER components are found at the edges of the cell, outside of the mitotic spindle zone. In these phases, reticular domains are absent and ER appears only as short, cisternae with ribosomes bound to the outer surfaces. We are currently working to characterize ER in the earliest (prophase and prometaphase) and latest (late anaphase and telophase) stages of mitosis. Studies of late telophase cells may also shed light on the early steps of nuclear envelope reformation. Immunofluorescence studies suggest an uneven distribution of certain ER markers and we anticipate that immunoelectron microscopy will be used to determine if cytoplasmic ER domains can be characterized on the molecular, as well as the structural, level.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 我们正在使用迅速冷冻的，冻结的非洲绿猴肾（BSC1）细胞的电子断层扫描来评估细胞分裂不同阶段内质网（ER）细胞良好结构的变化。 整个有丝分裂细胞的三维荧光显微镜研究表明，ER网络在有丝分裂的每个阶段都经历形态学变化，所有这些阶段都与相间ER模式不同。 电子断层扫描被用来补充LM工作，并将发现扩展到良好的结构水平。 到目前为止，我们已经准备好了9个间相细胞的层析成像数据集，中期中的19套细胞，3组后期和5组末期细胞。 最初的结果证实了在相间BSC1细胞中存在脑海中和网状ER结构域。 尽管这些结构域是连续的，但它们主要位于细胞质的不同部位。蓄水域更接近核，而网状区域靠近细胞周围。 在有丝分裂的过程中，最著名的是中期和后期，在细胞的边缘，在有丝分裂纺锤体外部的边缘发现了ER成分。 在这些阶段中，网状结构域不存在，ER似乎只有短的蓄水池，核糖体与外表面结合。 我们目前正在努力以最早的（预言和前期）以及有丝分裂的最新（后期和末期）阶段来表征ER。 对晚期末期细胞的研究还可以阐明核包膜改革的早期步骤。 免疫荧光研究表明，某些ER标记的分布不均匀，我们预计将使用免疫电子显微镜来确定是否可以在分子以及结构的水平上表征细胞质ER结构域。