Endoplasmic reticulum positioning and partitioning in mitotic HeLa cells

Cell division serves to distribute chromosomes and organelles into two daughter cells, but the mechanism of rough endoplasmic reticulum (RER) segregation in animal cell mitosis is poorly understood. Here we study the distribution of RER in mitotic HeLa cells and its relation to the cytoskeleton. At metaphase, the RER was located in the cell cortex and was most concentrated in two locations. Close to the plasma membrane the RER was closely associated with cortical actin, and after treatment with Latrunculin A RER elements retracted to the deep cortex and became more tubular. Positioning was therefore dependent on cortical F-actin. Deeper in the cortex cisternae were wrapped tightly around the contours of the spindle body and orientated along microtubules close the spindle poles. Stereology revealed a close correlation between RER volume and cell volume in telophase daughter cells. These results suggest that the RER is positioned at the outer and inner regions of metaphase cortex by association with cytoskeleton. This arrangement combined with a disposition in concentric layers, deep to the plasma membrane, appears to distribute the RER evenly in the cortex and may help to couple quantities of RER and cell constituents.