A role for local calcium gradients upon hypoxic injury in human umbilical vein endothelial cells (HUVEC)
| Autor: | Morito Monden, Tomio Kawasaki, Masataka Ikeda, Hideo Ariyoshi, Nobutoshi Shinoki, Norihide Yoshikawa, Jun-ichi Kambayashi, Masato Sakon |
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| Rok vydání: | 1998 |
| Předmět: |
Umbilical Veins
Cell Survival Physiology Analytical chemistry chemistry.chemical_element Biology Calcium Rhodamine 123 Umbilical vein Membrane Potentials chemistry.chemical_compound medicine Humans Viability assay Bleb (cell biology) Propidium iodide Molecular Biology Cell damage Cells Cultured Membrane potential Microscopy Confocal Intracellular Membranes Cell Biology medicine.disease Cell Hypoxia eye diseases Mitochondria chemistry Biophysics Endothelium Vascular Analog-Digital Conversion |
| Zdroj: | Cell Calcium. 24:49-57 |
| ISSN: | 0143-4160 |
| DOI: | 10.1016/s0143-4160(98)90088-4 |
| Popis: | Upon hypoxic injury, bleb formation is an early event of cell damage observed in a variety of cell types. Although a rise in cytosolic free Ca2+ ([Ca2+]i) has been considered to be involved in this process, the exact relationship between these phenomena remains ill-defined. In order to examine the relationship between bleb formation, and [Ca2+]i or nuclear free Ca2+ ([Ca2+]n), we analyzed [Ca2+]i and [Ca2+]n in HUVEC during hypoxic injury using confocal laser scanning microscopy. [Ca2+]i and [Ca2+]n were measured using Fluo-3, and cell viability and mitochondrial membrane potential were assessed by the exclusion of propidium iodide (PI) and rhodamine 123, respectively. After the initiation of hypoxia, [Ca2+]i and [Ca2+]n rose gradually up to 15 min reaching peak values of 447 +/- 62 and 516 +/- 105 nM, respectively, which was accompanied by a decrease in rhodamine 123 fluorescence and an increase in PI-stained cells. Bleb formation was observed after [Ca2+]i and [Ca2+]n had reached their peak values and the number of blebs increased thereafter. Confocal z-sectioning images revealed a localized increase in [Ca2+]i at the bleb forming site and this localized elevation in [Ca2+]i was observed before bleb formation in the corresponding area. In conclusion, bleb formation induced by hypoxic stress appears to involve Ca(2+)-dependent reactions that are linked to a regional elevation of [Ca2+]i. |
| Databáze: | OpenAIRE |
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