Ceramide inhibits the outward potassium current in rat pinealocytes

In the present study, we investigated the effect of ceramide on the outward K(+) current in rat pinealocytes using whole cell and single channel recordings. Three components of the whole cell outward K(+) current were separated, an iberiotoxin (IBTX)-sensitive K(+) current (I(KCa)), a transient A current (I(A)) and a delayed rectifier current (I(K)). C6-ceramide reduced all three components of the outward K(+) current. C6-ceramide (30 microM) caused a 53% inhibition of I(KCa) [a component that is generated by the IBTX-sensitive K(+) channel (BK channel)], a 27% inhibition of I(A) and a 17% inhibition of I(K). Additional studies showed that the BK channel was not inhibited by dihydroC6-ceramide, the inactive analog of C6-ceramide, but mimicked by sphingomyelinase which increased intracellular ceramide. The ceramide inhibition of the BK channel was only partly dependent on its inhibition of the L-type Ca(2+) channel. Studies using specific kinase inhibitors showed that calphostin C (a protein kinase C inhibitor) and to a lesser degree lavendustin A (a tyrosine kinase inhibitor) were effective in reducing the ceramide inhibition of I(KCa). Taken together, our results show that, in rat pinealocytes, ceramide reduces the outward K(+) current predominantly by inhibiting I(KCa). Moreover, protein kinase C appears to be the main kinase involved in the ceramide inhibition of I(KCa).