Phorbol ester-induced inhibition of potassium currents in rat sensory neurons requires voltage-dependent entry of calcium.
The whole cell patch-clamp technique was used to examine the effects of protein kinase C (PKC) activation (via the phorbol ester, phorbol 12,13 dibutyrate, PDBu) on the modulation of potassium currents (I(K)) in cultured capsaicin-sensitive neurons isolated from dorsal root ganglia from embryonic rat pups and grown in culture. PDBu, in a concentration- and time-dependent manner, reduced I(K) measured at +60 mV by approximately 30% if the holding potential (V(h)) was -20 or -47 mV but had no effect if V(h) was -80 mV. The PDBu-induced inhibition of I(K) was blocked by pretreatment with the PKC inhibitor bisindolylmaleimide I and I(K) was unaffected by 4-alpha phorbol, indicating that the suppression of I(K) was mediated by PKC. The inhibition of I(K) by 100 nM PDBu at a V(h) of -50 mV was reversed over several minutes if V(h) was changed to -80 mV. In addition, intracellular perfusion with 5 mM bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) or pretreatment with omega-conotoxin GVIA or Cd(2+)-Ringer, but not nifedipine, prevented the PDBu-induced suppression of I(K) at -50 mV, suggesting that a voltage-dependent influx of calcium through N-type calcium channels was necessary for the activation of PKC. The potassium channel blockers tetraethylammonium (TEA, 10 mM) and 4-aminopyridine (4-AP, 3 mM and 30 microM) reduced I(K), but only TEA attenuated the ability of PDBu to further inhibit the current, suggesting that the I(K) modified by PDBu was sensitive to TEA. Interestingly, in the presence of 3 mM or 30 microM 4-AP, 100 nM PDBu inhibited I(K) when V(h) was -80 mV. Thus 4-AP promotes the capacity of PDBu to reduce I(K) at -80 mV. We find that activation of PKC inhibits I(K) in rat sensory neurons and that voltage-dependent calcium entry is necessary for the development and maintenance of this inhibition.
Zhang
Y H
YH
Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, Indiana 46202-5120, USA.
Kenyon
J L
JL
Nicol
G D
GD
D015761:4-Aminopyridine / Q000494:pharmacology; D000704:Analysis of Variance; D000818:Animals; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005727:Ganglia, Spinal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D009475:Neurons, Afferent / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology*; D026902:Potassium Channel Blockers; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D019789:Tetraethylammonium / Q000494:pharmacology
D002121:Calcium Channel Blockers; D002614:Chelating Agents; D004791:Enzyme Inhibitors; D026902:Potassium Channel Blockers; D015240:Phorbol 12,13-Dibutyrate; D019789:Tetraethylammonium; D015761:4-Aminopyridine; D011493:Protein Kinase C; D002118:Calcium