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Journal of Neuroscience, Vol 15, 3307-3317, Copyright © 1995 by Society for Neuroscience
Modulation of the glutamate-evoked release of arachidonic acid from mouse cortical neurons: involvement of a pH-sensitive membrane phospholipase A2
N Stella, L Pellerin and PJ Magistretti
Laboratoire de Recherches Neurologiques, Faculte de Medecine, Universite de Lausanne, Switzerland.
Excitatory synaptic transmission is associated with changes in both
extracellular and intracellular pH. Using mouse cortical neurons in primary
cultures, we studied the sensitivity of glutamate-evoked release of
3H-arachidonic acid (3H-AA) to changes in extracellular pH (pHo) and
related intracellular pH (pHi). As pHo was shifted from 7.2 to 7.8, the
glutamate-evoked release of 3H-AA was enhanced by approximately threefold.
The effect of alkaline pHo on the glutamate response was rapid, becoming
significant within 2 min. 3H-AA release, evoked by both NMDA and kainate,
was also enhanced by pHo alkalinization. NMDA- and kainate-induced increase
in free intracellular Ca2+ was unaffected by changing pHo from 7.2 to 7.8,
indicating that the receptor-induced Ca2+ influx is not responsible for the
pHo sensitivity of the glutamate-evoked release of 3H-AA. Alkalinization of
pHi obtained by incubating neurons in the presence of HCO3- or NH4 enhanced
the glutamate-evoked release of 3H-AA, while pHi acidification obtained by
blockade of Na+/H+ and Cl-/HCO3- exchangers decreased the glutamate
response. Membrane-bound phospholipase A2 (mPLA2) activity was stimulated
by Ca2+ in a pH-dependent manner, increasing its activity as pH was shifted
from 7.2 to 7.8. This pH profile corresponds to the pH profile of the
glutamate-, NMDA- and kainate-evoked release of 3H-AA. Taken together,
these results indicate that the glutamate-evoked release of 3H-AA may be
mediated by the pH- sensitive mPLA2. Since excitatory neurotransmission
mediated by glutamate results in both pHo and pHi changes and since AA
enhances glutamatergic neurotransmission at both pre- and postsynaptic
levels, the data reported here reveals a possible molecular mechanism
whereby glutamate can modulate its own signalling efficacy in a
pH-dependent manner by regulating the release of AA.
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