Sequestration of iontophoretically injected calcium by living endothelial cells.
Sequestration of iontophoretically injected Ca2+ by monolayer culture cells (primary Xenopus laevis Tadpole Heart cells, XTH P, and an established cell line, XTH 2) is investigated. Injections are made at different velocities by changing the influx current. On Ca2+ injection the entire ER desintegrates, and near to the tip of the injecting pipette microtubules depolymerize. The time required to attain cell death is taken as the parameter indicating an overload of cellular Ca2+ sequestration capability. Three different Ca2+ transport kinetics are found: at Ca2+ flux rates of up to 20 X 10(-15) mol X s-1 (condition I) cells can tolerate long injection periods before they die; at flux rates from 20 to 40 X 10(-15) mol X s-1 (condition II) the injection time before cell death remains constant. Flux rates exceeding 40 X 10(-15) mol X s-1 decrease cellular Ca2+ sequestration capability to a minimum. These observations support the assumption of two Ca2+ sequestrating mechanisms: one of high affinity, but with low capacity (less than = 5 X 10(-15) mol X s-1) the other with low affinity for Ca2+ and a high capacity (10 to 40 X 10(-15) mol X s-1) for Ca2+ accumulation. Both mechanisms are saturable. As the Ca2+ sequestration velocity remains approximately constant in condition II, the capacity of the second mechanism seems to grow with increasing Ca2+ influx. The highly affin Ca2+ compartment is the ER, mitochondria form the less affin system. XTH 2 differ from primary cells by possessing a 5 to 8 fold higher Ca2+ sequestration capacity, whereas sequestration velocity is equal in both cell types.
D000818:Animals; D000968:Antimycin A / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D004727:Endothelium / Q000187:drug effects / Q000378:metabolism*; D007478:Iontophoresis; D007700:Kinetics; D008845:Microinjections; D008856:Microscopy, Fluorescence; D013050:Spectrometry, Fluorescence; D014982:Xenopus laevis
D000968:Antimycin A; D002118:Calcium