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Cell Swelling, Blebbing, and Death Are Dependent on ATP Depletion and Independent of Calcium During Chemical Hypoxia in a Glial Cell Line (ROC‐1)
Author(s) -
JurkowitzAlexander Marianne S.,
Altschuld Ruth A.,
Hohl Charlene M.,
Johnson J. David,
McDonald John S.,
Simmons Todd D.,
Horrocks Lloyd A.
Publication year - 1992
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1992.tb08910.x
Subject(s) - bleb (medicine) , calcium , programmed cell death , extracellular , calcium in biology , intracellular , chemistry , biophysics , biochemistry , biology , apoptosis , trabeculectomy , organic chemistry , neuroscience , glaucoma
The morphological and biochemical changes that occur during chemical hypoxic injury in a neural cell line were studied in the presence and absence of calcium. Oligo‐dendroglial‐glioma hybrid cells (ROC‐1) were subjected to inhibitors of glycolytic and oxidative ATP synthesis (chemical hypoxia). Complete respiratory inhibition depleted [ATP] to <5% of control by 4 min. Blebs appeared on the cell surfaces and cells began to swell within a few minutes of ATP depletion. A 200% increase in cell volume and bleb coalescence preceded irreversible cell injury (lactate dehy‐drogenase release) which began at ∼20 min with 50% cell death by 40 min. In energized cells an equivalent degree of osmotic swelling induced by ouabain inhibition of the Na + , K + ‐ATPase pump did not produce blebbing or cell death. Partial inhibition of respiration decreased [ATP] to ∼10% of control by 40 min. Blebbing and swelling began at 40 min and bleb coalescence preceded plasma membrane disruption which began at ∼55 min. ATP depletion, blebbing, swelling, and death followed similar time courses in the presence or absence of extracellular calcium ([Ca 2+ ] e ). Intracel‐lular calcium ([Ca 2+ ] i ) was measured using fura‐2. In calcium‐containing medium metabolic inhibition caused a transient increase in resting [Ca 2+ ] i (100 ± 17 n M ) followed by a low steady‐state level preceding plasma membrane disruption. Following deenergization in calcium‐free medium, [Ca 2+ ] i remained below 60 n M throughout injury and death. These data suggest that decreased ATP initiates a sequence of events including bleb formation and cell swelling that lead to irreversible cell injury in the absence of large increases in [Ca 2+ ] i .