A time-lapse video image intensification analysis of cytoplasmic organelle movements during endosome translocation.

作者：Benjamin A. Herman, David F. Albertini · 发表于：The Journal of Cell Biology · 年份：1984 · DOI：10.1083/jcb.98.2.565 · 被引用次数：157 · 研究领域：Photosynthetic Processes and Mechanisms、Advanced Fluorescence Microscopy Techniques、Cancer Treatment and Pharmacology

Vital fluorescence staining has been used in conjunction with time-lapse video image intensification microscopy to analyze the distribution and movement of endosomes, lysosomes, and mitochondria in cultured rat ovarian granulosa cells. Exposure of 5-d granulosa cell cultures to pyrene-concanavalin A (P-Con A) or 3,3'-dioctadecylindocarbocyanine-labeled low-density lipoprotein (dil-LDL) at 4 degrees C results in the formation of randomly distributed endosomes 10 min after warming to 37 degrees C that exhibit saltatory motion for 20 min. If granulosa cells are labeled at 4 degrees C with both P-Con A and dil-LDL and warmed to 37 degrees C, both ligands are found within the same endosomes which migrate centripetally to the cell center where label accumulates within phase-dense structures by 60 min. The initial endosome saltations occur over short distances (mean distance = 4.6 micron) with a mean velocity of 0.03 micron/s. Endosome saltations then cease and are followed by a gradual centriptal migration of endosomes to the cell center where they accumulate and fuse with phase-dense structures. The second phase of movement involves a continuous, unidirectional migration of endosomes over distances ranging from 5 to 40 micron at a mean velocity of 0.05 micron/s. Lysosomes were simultaneously visualized as acridine orange-staining, phase-dense structures in control cells and cells exposed to fluorescent ligands. In untreated cells, lysosomes are dispersed throughout the cytoplasm a...