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Gap‐junction blocker carbenoxolone differentially enhances NMDA‐induced cell death in hippocampal neurons and astrocytes in co‐culture
Author(s) -
Zündorf Gregor,
Kahlert Stefan,
Reiser Georg
Publication year - 2007
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.1471-4159.2007.04509.x
Subject(s) - carbenoxolone , depolarization , gap junction , nmda receptor , astrocyte , membrane potential , excitotoxicity , biology , chemistry , neuron , microbiology and biotechnology , biophysics , neuroscience , intracellular , biochemistry , central nervous system , receptor
The beneficial or detrimental role of gap junction communication in the pathophysiology of brain injury is still controversial. We used co‐cultures of hippocampal astrocytes and neurons, where we identified homocellular astrocyte–astrocyte and heterocellular astrocyte–neuron coupling by fluorescence recovery after photobleaching, which was decreased by the gap junction blocker carbenoxolone (CBX). In these cultures, we determined the cell type‐specific effects of CBX on the excitotoxic damage caused by N ‐methyl‐ d ‐aspartate (NMDA). We determined in both astrocytes and neurons the influence of CBX, alone or together with NMDA challenge, on cytotoxicity using propidium iodide labeling. CBX alone was not cytotoxic, but CBX treatment differentially accelerated the NMDA‐induced cell death in both astrocytes and neurons. In addition, we measured mitochondrial potential using rhodamine 123, membrane potential using the oxonol dye bis(1,3‐diethylthiobarbituric acid)trimethine oxonol, cytosolic Ca 2+ level using fura‐2, and formation of reactive oxygen species (ROS) using dihydroethidium. CBX alone induced neither an intracellular Ca 2+ rise nor a membrane depolarization. However, CBX elicited a mitochondrial depolarization in both astrocytes and neurons and increased the ROS formation in neurons. In contrast, NMDA caused a membrane depolarization in neurons, coinciding with intracellular Ca 2+ rise, but neither mitochondrial depolarization nor ROS production seem to be involved in NMDA‐mediated cytotoxicity. Pre‐treatment with CBX accelerated the NMDA‐induced membrane depolarization and prevented the repolarization of neurons after the NMDA challenge. We hypothesize that these effects are possibly mediated via blockage of gap junctions, and might be involved in the mechanism of CBX‐induced acceleration of excitotoxic cell death, whereas the CBX‐induced mitochondrial depolarization and ROS formation are not responsible for the increase in cytotoxicity. We conclude that both in astrocytes and neurons gap junctions provide protection against NMDA‐induced cytotoxicity.