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Can J Physiol Pharmacol ; 93(4): 291-7, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25741585

RESUMEN

The objective of this work was to verify whether, as in the case of the plasma membrane of human vascular smooth muscle cells (hVSMCs), cytosolic ET-1-induced increase of nuclear calcium is mediated via the activation of calcium influx through the steady-state R-type calcium channel. Pharmacological tools to identify the R-type calcium channels, as well as real 3-D confocal microscopy imaging techniques coupled to calcium fluorescent probes, were used to study the effect of cytosolic ET-1 on nuclear calcium in isolated nuclei of human hepatocytes and plasma membrane perforated hVSMCs. Our results showed that pre-treatment with pertussis toxin (PTX) or cholera toxin (CTX) prevented cytosolic ET-1 (10(-9) mol/L) from inducing a sustained increase in nuclear calcium. Furthermore, the L-type calcium channel blocker nifedipine did not prevent cytosolic ET-1 from inducing an increase in nuclear calcium, as opposed to the dual L- and R-type calcium channel blocker isradipine (PN200-110) (in the presence of nifedipine). In conclusion, the preventative effect with PTX and CTX, and the absence of an effect with nifedipine, as well as the blockade by isradipine on cytosolic ET-1-induced increase in nuclear calcium, suggest that this nuclear calcium influx in hVSMCs is due to activation of the steady-state R-type calcium channel. The sarcolemmal and nuclear membrane R-type calcium channels in hVSMCs are involved in ET-1 modulation of vascular tone in physiology and pathology.


Asunto(s)
Canales de Calcio Tipo R/metabolismo , Señalización del Calcio , Proteínas de Transporte de Catión/metabolismo , Núcleo Celular/metabolismo , Citosol/metabolismo , Endotelina-1/metabolismo , Músculo Liso Vascular/metabolismo , Membrana Nuclear/metabolismo , Aorta , Bloqueadores de los Canales de Calcio/farmacología , Canales de Calcio Tipo R/química , Señalización del Calcio/efectos de los fármacos , Proteínas de Transporte de Catión/química , Permeabilidad de la Membrana Celular/efectos de los fármacos , Núcleo Celular/efectos de los fármacos , Células Cultivadas , Toxina del Cólera/farmacología , Citosol/efectos de los fármacos , Hepatocitos/citología , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Humanos , Imagenología Tridimensional , Fusión de Membrana/efectos de los fármacos , Moduladores del Transporte de Membrana/farmacología , Microscopía Confocal , Músculo Liso Vascular/citología , Músculo Liso Vascular/efectos de los fármacos , Membrana Nuclear/efectos de los fármacos , Toxina del Pertussis/farmacología , Sarcolema/efectos de los fármacos , Sarcolema/metabolismo
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