GABA INDUCES CA2+ TRANSIENTS IN ASTROCYTES

By using the Ca2+-sensitive indictor Fura-2/AM, the cytosolic Ca2+ levels [Ca2+]i were measured in type 1 astrocytes in rat cortical astroglial primary cultures, after stimulation with GABA, muscimol (GABA(A) agonist), or baclofen (GABA(B) agonist). We report the first evidence that stimulation of both GABA(A) and GABA(B) receptors evokes Ca2+ transients in type 1 astrocytes. Two types of Ca2+ responses were seen: the single-phase curve, which was the most common, and the biphasic, which consisted of an initial rise that persisted at the maximal or submaximal level. Both types of Ca2+ responses appeared with some latency. The responses were obtained in astrocytes grown for 12-16 days in culture and the response frequencies for all three agonists were 18% of the total number of examined cells. However, when the astrocytes were grown in a mixed astroglial/neuronal culture the response frequencies for all three agonists increased to 35% of the total number of examined cells. In some cells, the responses after GABA stimulation were blocked to baseline levels after exposure to bicuculline (GABA(A) antagonist). In other cells, bicuculline only slightly reduced the GABA-evoked responses, and the addition of phaclofen (GABA(B) antagonist) did not potentiate this partial inhibition. However, the muscimol-evoked rises in [Ca2+], were completely inhibited after exposure to bicuculline, while the responses after baclofen could only be partly blocked by phaclofen. GABA evoked rises in [Ca2+]i which alternatively were inhibited (mostly) or persisted in Ca2+-free buffer. The rises in [Ca2+]i persisted, but were reduced, in Ca2+-free buffer after stimulation with muscimol, but were inhibited after baclofen stimulation. The GABA uptake blockers guvacine, 4,5,6,7-tetrahydroisoxazolo(4,5-c)pyridin-3-ol and nipecotic acid were also able to reduce the GABA-evoked rises in [Ca2+]i. However, the L-type Ca2+ channel antagonist nifedipine failed to influence on the GABA-evoked Ca2+ transients. The results suggest that type 1 astrocytes in primary culture express GABA receptors which can elevate [Ca2+], directly or indirectly via Ca2+ Channels and/or via release from internal Ca2+ stores. The results also suggest that GABA can have intracellular Ca2+-mobilizing sites since the GABA-evoked responses were reduced after incubation with GABA uptake blockers.