Modulation of voltage-dependent Ba2+ currents in the guinea-pig gastric antrum by cyclic nucleotide-dependent pathways

摘要

class="enumerated" style="list-style-type:decimal">We have investigated whether the activation of cAMP- and cGMP-dependent pathways modifies the properties of voltage-dependent Ba²⁺ currents (IBa) recorded from guinea-pig gastric myocytes using patch-clamp techniques. All experiments were carried on single smooth muscle cells, dispersed from the circular layer of the guinea-pig gastric antrum.Both dibutyryl cAMP (db-cAMP, 0.1–1 mM), a membrane-permeable ester of cAMP, and isoproterenol, a selective β-stimulant, inhibited IBa in a concentration-dependent manner.Forskolin, but not dideoxy-forskolin, an inactive isomer of forskolin, inhibited the peak amplitude of IBa.In the presence of either Rp-cAMP or the PKA (cAMP-dependent protein kinase) inhibitor peptide 5-24 (PKA-IP), neither forskolin nor db-cAMP inhibited IBa.After establishing a conventional whole-cell recording, the peak amplitude of IBa gradually decreased when the catalytic subunit of PKA was included in the pipette. The further application of Rp-cAMP reversibly enhanced IBa.Sodium nitroprusside (0.1–1 mM) and 8-Br-cGMP (0.1–1 mM) also inhibited IBa in a concentration-dependent manner.The inhibitory effects of forskolin or db-cAMP on IBa were not significantly changed by pretreatment with a cGMP-dependent protein kinase (PKG) inhibitor. Similarly, the inhibitory actions of 8-Br-cGMP on IBa were not modified by PKA-IP.The membrane-permeable cyclic nucleotides db-cAMP and 8-Br-cGMP caused little shift of the voltage dependence of the steady-state inactivation and reactivation curves.Neither of the membrane-permeable cyclic nucleotides db-cAMP or 8-Br-cGMP had additive inhibitory effects on IBa.These results indicate that two distinct cyclic nucleotide-dependent pathways are present in the guinea-pig gastric antrum, and that both inhibited IBa in an independent manner.