Abstract:
:Neurotransmitter release and changes in the concentration of intracellular free calcium ([Ca++]i) were studied in cultured GABAergic cerebral cortical neurons, from mice, upon depolarization with either an unphysiologically high potassium concentration (55 mM) or the physiological excitatory neurotransmitter glutamate (100 microM). Both depolarizing stimuli exerted prompt increases in the release of preloaded [3H]GABA as well as in [Ca++]i. However, the basic properties of transmitter release and the increase in [Ca++]i under a variety of conditions were different during stimulation with K+ or glutamate. Potassium-evoked release of [3H]GABA consisted of two phases, a rapid, large and transient phase followed by a smaller, more persistent second phase. The rapid phase was inhibited (60%) by nocodazole which reduced the number of vesicles in the neurites by 80%. This rapid phase of the GABA release was also reduced by organic (verapamil) and inorganic (Co++) Ca++ channel blockers but was insensitive to the GABA transport inhibitor SKF 89976A. In contrast, the second phase was less sensitive to nocodazole and Ca++ channel antagonists but could be inhibited by SKF 89976A. The glutamate-induced [3H]GABA release, which was mainly mediated by N-methyl-D-aspartate receptors, consisted of a single, sustained phase. This was insensitive to nocodazole, partly inhibited by verapamil and could be blocked by Co++ as well as SKF 89976A. The action of Co++ could be attributed to a block of N-methyl-D-aspartate-associated ion channels. These findings strongly suggest that the majority of the K(+)-stimulated GABA release is dependent upon vesicles whereas the glutamate induced release is non-vesicular and mediated by a depolarization-dependent reversal of the direction of high-affinity GABA transport. The basic differences in the mode of action of the two depolarizing stimuli were reflected in the properties of the increase in [Ca++]i elicited by 55 mM K+ and 100 microM glutamate, respectively. The K(+)-induced increase in [Ca++]i was reduced by both verapamil and Ca(++)-free media whereas the corresponding glutamate response was only sensitive to Ca(++)-free conditions. Exposure of the cells to nocodazole or SKF 89976A had no effect on the ability of K+ or glutamate to increase [Ca++]i. Altogether, the results clearly demonstrate that K(+)-induced transmitter release from these GABAergic neurons is vesicular in nature whereas that induced by the neurotransmitter glutamate is not.
journal_name
Neurosciencejournal_title
Neuroscienceauthors
Belhage B,Hansen GH,Schousboe Adoi
10.1016/0306-4522(93)90592-4subject
Has Abstractpub_date
1993-06-01 00:00:00pages
1019-34issue
4eissn
0306-4522issn
1873-7544pii
0306-4522(93)90592-4journal_volume
54pub_type
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