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Formation and disintegration of cisternae of the endoplasmic reticulum visualized in live cells by conventional fluorescence and confocal laser scanning microscopy: Evidence for the involvement of calcium and the cytoskeleton

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Summary

The influence of substances interfering with the cellular calcium distribution on the organization of the endoplasmic reticulum has been investigated in live epidermal cells of onion bulb scales. The endoplasmic reticulum was visualized by vital staining with the fluorochrome DiOC6(3). It constitutes in these cells an anastomosing membrane system which is composed of three forms: cisternae, short tubules forming a peripheral network, and long tubular strands deeper in the cytoplasm. In the presence of all tested calcium interfering substances, e.g. the ionophore calcimycin (5 μM), the cryptate 221 (0.5 mM), the calmodulin antagonist calmidazolium (10 μM), the tubular ER elements disappear and huge cisternae form instead. The potassium-selective cryptate 222 (1 mM) chemically very similar to the effective cryptate 221 does not cause this change in ER pattern. Actin filaments which are indispensable for ER distribution in the epidermis cells appear to fragment in the presence of the drugs indicating some similarity with the action of cytochalasin D (Quader et al. 1989). Removal of the drugs initiates a characteristic sequence of recovery. The cisternae disintegrate at their edges into tubular loops which are pulled away from this cellular site as long tubular strands. In the presence of cytochalasin D (2 μM) the disintegration of the cisternae is inhibited indicating that kinetic forces are necessary to generate and maintain the spatial distribution of at least parts of the tubular ER meshwork. For the first time the decay of cisternae is described in live cells. The effect of the calcium agents is also compared with changes in ER organization caused by other chemical or natural means.

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Abbreviations

ER:

endoplasmic reticulum

EGTA:

ethyleneglycol-bis-(β-aminoethyl-ether)-tetraacetic acid

DiOC6(3):

3,3′-dihexylox-acarbocyanin-iodide

CLSM:

confocal laser scanning microscopy

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  1. Zellenlehre, Fakultät für Biologie, Universität Heidelberg, Heidelberg

    H. Quader

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Quader, H. Formation and disintegration of cisternae of the endoplasmic reticulum visualized in live cells by conventional fluorescence and confocal laser scanning microscopy: Evidence for the involvement of calcium and the cytoskeleton. Protoplasma 155, 166–175 (1990). https://doi.org/10.1007/BF01322626

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