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Multiple voltage-sensitive calcium channels are probably involved in endogenous GABA release from striatal neurones differentiated in primary culture

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Summary

Calcium-dependent release of neurotransmitters is thought to be due to Ca2+ entry into nerve terminals, but the identities of the various voltage-sensitive Ca2+ channels (VSCC) involved in this process remain obscure. To elucidate the types of VSCCs involved in the release process, we studied the effects of various organic Ca2+ channel antagonists and agonists on the release of endogenous γ-aminobutyric acid (GABA) from mouse striatal neurones differentiated in primary culture. Diltiazem, verapamil and methoxyverapamil (D 600) inhibited K+-evoked (30 mM) GABA release at very high concentrations (> 1 μM). The dihydropyridine (DHP) nifedipine, at low concentrations (0.01–1.00 μM), was able to inhibit part of the K+-evoked GABA release (25.6±7.3% inhibition at 1 μM). This is in agreement with the high affinity of nifedipine for DHP binding sites. The DHPs, BAY K 8644 (EC50 = 41±15 nM) and CGP 28.392, which possess agonist properties at VSCCs, increased the 15 mM K+-evoked GABA release. The release evoked by the combination of K+ (15 μM) and BAY K 8644 (up to 10 μM) remained smaller than the release elicited by 30 mM K+. The effect of BAY K 8644 (1 μM) was inhibited by nifedipine (IC50 0.55±0.05 μM). When Na+ ions were replaced by choline, basal and K+-evoked GABA release was significantly increased. Even in the absence of external Na+, nifedipine (1 μM) was not able to totally block the K+ effect. Moreover amiloride, a drug known to inhibit Na+/Ca+ exchange, and tetrodotoxin (TTX), did not modify the 30 mM K+ response. Therefore, nifedipine-insensitive K+-evoked GABA release is not due to Na+-dependent Ca2+ entry. These results can be explained by the presence of DHP-sensitive and insensitive Ca2+ channels on nerve terminals, each involved in the release process.

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Abbreviations

CGP 28.392:

4-[2-(difluoromethoxy)phenyl]-1,4,5,7-tetrahydro-2-methyl-5-oxo-furo[3,4-b]pyridine-3-carboxylic acid ethylester

BAY K 8644:

(±)-methyl 1,4-dihydro-2,6-dimethyl-3nitro-4-(2 trifluoromethyl) pyridine-5-carboxylate

DHP:

1,4-dihydropyridine

DIV:

days in vitro

D 600:

methoxyverapamil

GABA:

γ-aminobutyric acid

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

HPLC:

high performance liquid chromatography

TTX:

tetrodotoxin

VSCC:

voltage-sensitive calcium channel

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  1. Centre CNRS, INSERM de Pharmacologie, Endocrinologie, Rue de la Cardonille, F-34094, Montpellier Cedex, France

    Jean-Philippe Pin & Jöel Bockaert

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  1. Jean-Philippe Pin
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Pin, JP., Bockaert, J. Multiple voltage-sensitive calcium channels are probably involved in endogenous GABA release from striatal neurones differentiated in primary culture. Naunyn-Schmiedeberg's Arch Pharmacol 336, 190–196 (1987). https://doi.org/10.1007/BF00165804

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  • DOI: https://doi.org/10.1007/BF00165804

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