Distinct protein kinases regulate SNAP-25 expression in chromaffin cells

Montiel, C; Mendoza, I.; Garcia, CJ; Awad, Y; Garcia-Olivares, J; Solis-Garrido, LM; Lara, H; Garcia, AG; Cárdenas AM

Abstract

The contribution of distinct Ca2+-sensitive protein kinases to the regulation of the expression of the synaptosomal-associated protein SNAP-25 was examined in bovine chromaffin cells. Prolonged incubation with high K+ (38 mM) or 1,1-dimethyl-4-phenyl-piperazinium (DMPP), a nicotinic receptor agonist, significantly increased SNAP-25 protein and mRNA expression, as assessed by immunoblotting and semi-quantitative RT-PCR analysis. Both stimuli preferentially enhanced mRNA coding for the SNAP-25a isoform. Increase of SNAP-25 expression induced by K+ or DMPP was inhibited over 70% by KN-62 and KN-93, two Ca2+/calmodulin-dependent protein kinase (CaMK) inhibitors, whereas the inactive analogue KN-92 only reduced the expression by 34%. The three compounds also inhibited the high K+-elicited [Ca2+]i signal by 40%, suggesting that the effect of KN-62 and KN-93 was a combination of CaMK/Ca2+ influx inhibitory actions. Incubation of the cells with mitogen-activated protein kinase (MAPK) inhibitors PD98059 and U0126 reduced protein expression elicited by high K+ by 50%, but did not modify the response to DMPP. Interestingly, although protein kinase A (PKA) inhibition by H-89 did not affect the high K+ or DMPP-induced SNAP-25 expression, basal protein levels were significantly modified upon activation or inhibition of this pathway. Basal expression of SNAP-25 was also modified by the protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate, but not by Gö6976, a PKC-? inhibitor, suggesting that the Ca2+-insensitive PKC-? isoform control basal expression of SNAP-25 in these cells. Taken together, these results provide the first evidence that diverse protein kinases might converge in the induction of SNAP-25 expression in chromaffin cells. The preferential contribution of one or another kinase would depend on the physiological or experimental conditions. © 2002 Wiley-Liss, Inc.

Más información

Título según WOS: Distinct protein kinases regulate SNAP-25 expression in chromaffin cells
Título según SCOPUS: Distinct protein kinases regulate SNAP-25 expression in chromaffin cells
Título de la Revista: JOURNAL OF NEUROSCIENCE RESEARCH
Volumen: 71
Número: 3
Editorial: Wiley
Fecha de publicación: 2003
Página de inicio: 353
Página final: 364
Idioma: English
URL: http://doi.wiley.com/10.1002/jnr.10499
DOI:

10.1002/jnr.10499

Notas: ISI, SCOPUS