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Naphthalenesulfonamide derivatives ML9 and W7 inhibit catecholamine secretion in intact and permeabilized chromaffin cells

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Abstract

The role of protein phosphorylation in catecholamine secretion from bovine adrenomedullary chromaffin cells was studied using different protein kinase inhibitors. Naphthalenesulfonamide derivatives as ML9 and ML7, more specific for the myosin light chain kinase, and the calmodulin antagonist W7 inhibited catecholamine secretion 20 and 40% respectively in digitonin-permeabilized chromaffin cells. ML9 also decreased calcium evoked protein phosphorylation of different proteins including tyrosine hydroxylase in permeabilized cells. These naphthalenesulfonamide derivatives showed also an effect in intact cells, ML9 and W7 produced 50% inhibition in catecholamine secretion and45Ca2+ uptake, however H8 had no effect. The partial [3H]nitrendipine binding displacement of these drugs to adrenomedullary membranes suggests that these sulfonamide derivatives could interact directly with L-type calcium channels in intact cells. The results obtained in permeabilized cells suggest a possible role of protein phosphorylation in the regulation of catecholamine secretion in chromaffin cells.

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Abbreviations

ML9:

1-(5-Chloronaphthalene-1-sulfonyl)1H-hexahydro-1,4-diazepine hydrochloride

ML7:

1-(5-Iodonaphthalene-1-sulfonyl)-1H-hexahydro-1,4 diazepine hydrochloride

H7:

1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride

H8:

N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride

W7:

N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride

PKI:

protein kinase A inhibitor

HEPES:

N-(2-hydroxyethylpiperazine-N′-(2 ethanesulfonic acid)

PIPES:

piperazine-N, N′-bis (2-ethanesulfonic acid)

EGTA:

[ethylene-bis (oxyethylenenitrilo)] tetraacetic acid

SDS:

sodium dodecyl sulphate

PAGE:

polyacrylamide gel electrophoresis

DMEM:

Dulbecco's Modified Eagle's medium

MLC:

myosin light chain

MLCK:

myosin light chain kinase

TH:

tyrosine hydroxylase

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Authors and Affiliations

  1. Grupo de Farmacologia Bioquimica, Departamento de Neuroquimica, Facultad de Medicina. Instituto de Neurociencias, Universidad de Alicante, Ap. 374. 03080, Alicante, Spain

    J. A. Reig, S. Viniegra, J. J. Ballesta, M. Palmero & L. M. Gutierrez

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  1. J. A. Reig
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  2. S. Viniegra
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  3. J. J. Ballesta
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  4. M. Palmero
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  5. L. M. Gutierrez
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Reig, J.A., Viniegra, S., Ballesta, J.J. et al. Naphthalenesulfonamide derivatives ML9 and W7 inhibit catecholamine secretion in intact and permeabilized chromaffin cells. Neurochem Res 18, 317–323 (1993). https://doi.org/10.1007/BF00969089

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