Nitric oxide (NO; 1 μM) or an NO donor (500 μM diethylenetriamine‐nitric oxide, DETA‐NONOate) caused rapid glutamate and ATP release from cultured rat cortical astrocytes. NO‐induced glutamate release was prevented by calcium chelators (EGTA or BAPTA‐AM) and an inhibitor of vesicular exocytosis (botulinum neurotoxin C, BoTx‐C), but not by a glutamate transport inhibitor, L‐trans‐pyrrolidine‐2,4‐dicarboxylate (t‐PDC), a cyclooxygenase inhibitor (indomethacin), or an inhibitor of soluble guanylate cyclase 1H‐(1,2,4)oxadiazolo‐(4,3‐a)quinoxalin‐1‐one (ODQ), and was not induced by mitochondrial respiratory inhibitors (myxothiazol or azide). Similarly to glutamate, NO‐induced ATP release was also completely blocked by BAPTA‐AM and BoTx‐C, suggesting again a vesicular, calcium‐dependent mechanism of release. Addition of DETA‐NONOate (500 μM) to fura‐2–loaded astrocytes induced a rapid, transient increase in intracellular calcium levels followed by a lower, sustained level of calcium entry. The latter was blocked by gadolinium (1 μM), an inhibitor of capacitative Ca2+ entry. Thus, NO appears to cause rapid exocytosis of vesicular glutamate and ATP from astrocytes by raising intracellular calcium levels. Astrocytes activated by lipopolysaccharide/endotoxin and interferon‐γ to express inducible NO synthase (iNOS) maintained substantially higher extracellular glutamate levels than nonactivated cells or activated cells treated with an iNOS inhibitor (1400W), but the rate of glutamate uptake by these cells was similar. This suggests that NO from inflammatory‐activated astrocytes causes release of astrocytic glutamate. NO‐induced release of astrocytic glutamate and ATP may be important in physiological or pathological communication between astrocytes and neurons. GLIA 40:312–323, 2002. © 2002 Wiley‐Liss, Inc.
Glia – Wiley
Published: Dec 1, 2002
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