Cytosolic free magnesium, ATP, and blebbing during chemical hypoxia in cultured rat hepatocytes

Andrew W. Harman, Anna Liisa Nieminen, John J. Lemasters, Brian Herman

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Abstract

Cytosolic free Mg2+ concentration was determined in 1-day cultured rat hepatocytes using Multiparameter Digitized Video Microscopy (MDVM) of the fluorescent probe, magfura-2. Chemical hypoxia with KCN (5 mM) and iodoacetate (1 mM), a model which mimics the ATP depletion and reductive stress of hypoxia, caused a rapid increase of free Mg2+ from 1.1 ± 0.2 to 1.6 ± 0.2 mM within 4 min. Concurrently, numerous small plasma membrane blebs formed and ATP levels dropped from 13.24 to 1.32 nmol/106 cells. Removal of KCN and iodoacetate resulted in recovery of ATP to 60-70% of pre-exposure levels, a concomitant decrease in cytosolic free Mg2+ back toward basal levels, and reversal of blebbing (bleb resorption). These results indicate that changes of cytosolic free Mg2+ inversely reflect changes of ATP in a model of hypoxia and reoxygenation. Bleb formation and resorption were dependent on the fall and rise of ATP.

Original languageEnglish (US)
Pages (from-to)477-483
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume170
Issue number2
DOIs
StatePublished - Jul 31 1990

Bibliographical note

Funding Information:
r This work was supported by Grants AGO7218 and DK30874 from the National Institutes of Health, the Gustavas and Louise Pfeiffer Foundation, and Grant J-1433 from the Office of Naval Research.

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