Enhancement of Ca2+ channel currents in human neuroblastoma (SH-SY5Y) cells by phorbol esters with and without activation of protein kinase ... View Full Text


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Article Info

DATE

1995-03

AUTHORS

Helen L. Reeve, Peter F. T. Vaughan, Chris Peers

ABSTRACT

The effects of phorbol esters on Ca2+ channel currents in human neuroblastoma SH-SY5Y cells were studied using whole-cell patch-clamp recordings. Bath application of 12-O-tetradecanoylphorbol-13-acetate (TPA) or phorbol 12,13-dibutyrate (PDBu; 100 nM to 1 μM), known activators of protein kinase C (PKC), enhanced Ca2+ channel currents in a voltage-dependent manner similar to that of Bay K 8644. TPA also enhanced Ca2+ channel currents during cell dialysis with the PKC pseudosubstrate, PKC(19–36), and in cells which had been pre-incubated with 500 nM staurosporine, and which were exposed to staurosporine during recordings. Application of 4α-phorbol12, 13-didecanoate (4α-PDD; 100 nM), which does not activate PKC, caused current enhancement similar to the effects of TPA. However, intracellular dialysis of TPA was without effect on Ca2+ channel currents. Residual Ca2+ channel currents recorded after exposure to 1 μM ω-conotoxin GVIA were still enhanced by TPA, but in the presence of either Bay K 8644 (5 μM) or nifedipine (5 μM), TPA was without effect. When cells were pre-incubated for 10 min at 37° C with 100 nM TPA, currents subsequently recorded in its absence were enhanced as compared to untreated cells; 5 μM nifedipine still inhibited currents to the same degree. This enhancement was not mimicked by 4αPDD, and was inhibited by staurosporine. Our results indicate that acute applications of phorbol esters (at concentrations commonly used to activate PKC) enhance L-type Ca2+ channel currents in SH-SY5Y cells via a PKC-independent mechanism which appears similar to that induced by Bay K 8644. By contrast, pre-incubation with TPA enhances both L- and N-type currents via activation of PKC. More... »

PAGES

729-737

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf00373996

DOI

http://dx.doi.org/10.1007/bf00373996

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1018064352

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/7540748


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