Ultrastructural analysis of mitotic Drosophila S2 cells identifies distinctive microtubule and intracellular membrane behaviors.

摘要: S2 cells are one of the most widely used Drosophila melanogaster cell lines. A series studies has shown that they particularly suitable for RNAi-based screens aimed at dissection cellular pathways, including those controlling shape and motility, metabolism, host–pathogen interactions. In addition, RNAi in been successfully to identify many new mitotic genes conserved higher eukaryotes, analysis several aspects process. However, no detailed complete description mitosis ultrastructural level done. Here, we provide a characterization all phases visualized by transmission electron microscopy (TEM). We analyzed TEM random sample 144 undergoing mitosis, focusing on intracellular membrane microtubule (MT) behaviors. This unbiased approach provided comprehensive view dividing cells, allowed us discover exhibit previously uncharacterized behavior membranes, involving formation quadruple nuclear early prometaphase its disassembly during late prometaphase. After envelope disassembly, apparatus becomes encased discontinuous network endoplasmic reticulum which associate with mitochondria, presumably prevent their diffusion into spindle area. also observed peculiar metaphase organization. found kinetochores attached k-fibers almost invariably associated lateral MT bundles can be either interpolar or connected different kinetochore. organization is likely favor chromosome alignment subsequent segregation anaphase. discovered unknown features mitosis. The genetic determinants these now investigated, instance using an approach, easy efficient cells.