A target K+ channel for the LP-805-induced hyperpolarization in smooth muscle cells of the rabbit portal vein.

The resting membrane potential of smooth muscle cells of the rabbit portal vein was -51.2 mV. LP-805 (8-tert-butyl-6,7-dihydropyrrolo[3,2-e] 5-methylpyrazolo [1,5-a] pyrimidine-3-carbonitrile) hyperpolarized the membrane to -62.3 mV (10 microM) and inhibited the burst spike discharges as measured using the microelectrode method. In dispersed smooth muscle cells, LP-805 (10 microM) generated an outward-current with a maximum amplitude of 68 pA at a holding potential of -40 mV in experiments using the voltage-clamp procedure. The reversal potential of the outward current evoked by LP-805 was -82 mV and this value was close to the equilibrium potential for K+ (-80 mV) in the present ionic conditions, suggesting that LP-805 activated the K+ channel. Generation of both the hyperpolarization and the outward current by LP-805 was inhibited by glibenclamide (> or = 1 microM). Using the cell-attached and cell-free patch-clamp (in the presence of GDP) procedures, the maxi-K+ channel current (150 pS) could be recorded in the absence of LP-805; application of LP-805 additionally opened a small conductance K+ channel current (15 pS) without change in the activity of the maxi-K+ channel. The maxi-K+ channel was sensitive to charybdotoxin (0.1 microM) and to intracellular Ca2+ ([Ca2+]i) concentration. The 15 pS channel was insensitive to [Ca2+]i and charybdotoxin, but sensitive to intracellular ATP concentration. Glibenclamide (> 1 microM) inhibited the 15 pS K+ channel activated by LP-805.(ABSTRACT TRUNCATED AT 250 WORDS)