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

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

doi:10.1016/0006-291X(90)92116-H

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

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

Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

76 Scopus citations

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 language	English (US)
Pages (from-to)	477-483
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	170
Issue number	2
DOIs	https://doi.org/10.1016/0006-291X(90)92116-H
State	Published - 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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title = "Cytosolic free magnesium, ATP, and blebbing during chemical hypoxia in cultured rat hepatocytes",

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.",

author = "Harman, \{Andrew W.\} and Nieminen, \{Anna Liisa\} and Lemasters, \{John J.\} and Brian Herman",

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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AU - Harman, Andrew W.

AU - Nieminen, Anna Liisa

AU - Lemasters, John J.

AU - Herman, Brian

N1 - 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.

PY - 1990/7/31

Y1 - 1990/7/31

N2 - 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.

AB - 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.

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

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