title: Demonstration of an inwardly rectifying K+ current component modulated by thyrotropin-releasing hormone and caffeine in GH3 rat anterior pituitary cells.
abstract: Reduction of an inwardly rectifying K+ current by thyrotropin-releasing hormone (TRH) and caffeine has been considered to be an important determinant of electrical activity increases in GH3 rat anterior pituitary cells. However, the existence of an inwardly rectifying K+ current component was recently regarded as a misidentification of an M-like outward current, proposed to be the TRH target in pituitary cells, including GH3 cells. In this report, an inwardly rectifying component of K+ current is indeed demonstrated in perforated-patch voltage-clamped GH3 cells. The degree of rectification varied from cell to cell, but both TRH and caffeine specifically blocked a fraction of current with strong rectification in the hyperpolarizing direction. Use of ramp pulses to continuously modify the membrane potential demonstrated a prominent blockade even in cells with no current reduction at voltages at which M-currents are active. Depolarization steps to positive voltages at the maximum of the inward current induced a caffeine-sensitive instantaneous outward current followed by a single exponential decay. The magnitude of this current was modified in a biphasic way according to the duration of the previous hyperpolarization step. The kinetic characteristics of the current are compatible with the possibility that removal from inactivation of a fast-inactivating delayed rectifier causes the hyperpolarization-induced current. Furthermore, the inwardly rectifying current was blocked by astemizole, a potent and selective inhibitor of human ether-á-go-go -related gene (HERG) K+ channels. Along with other pharmacological and kinetic evidence, this indicates that the secretagogue-regulated current is probably mediated by a HERG-like K+ channel. Addition of astemizole to current-clamped cells induced clear increases in the frequency of action potential production. Thus, an inwardly-rectifying K+ current and not an M-like outward current seems to be involved in TRH and caffeine modulation of electrical activity in GH3 cells.
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Oviedo, E-33006, Oviedo, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Barros
authors.forename: D
authors.initials: D
authors.lastname: del Camino
authors.forename: L A
authors.initials: LA
authors.lastname: Pardo
authors.forename: T
authors.initials: T
authors.lastname: Palomero
authors.forename: T
authors.initials: T
authors.lastname: Giráldez
authors.forename: P
authors.initials: P
authors.lastname: de la Peña
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D016589:Astemizole / Q000494:pharmacology; D002110:Caffeine / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D027682:Cation Transport Proteins; D002460:Cell Line; D004268:DNA-Binding Proteins; D000072237:ERG1 Potassium Channel; D004553:Electric Conductivity; D051638:Ether-A-Go-Go Potassium Channels; D006801:Humans; D007700:Kinetics; D008564:Membrane Potentials; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000502:physiology*; D015221:Potassium Channels / Q000187:drug effects* / Q000502:physiology*; D024642:Potassium Channels, Voltage-Gated; D051381:Rats; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology*; D015534:Trans-Activators; D000071230:Transcriptional Regulator ERG
chemical_list: D027682:Cation Transport Proteins; D004268:DNA-Binding Proteins; C072312:ERG protein, human; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; C000606913:KCNH2 protein, human; C092509:KCNH6 protein, human; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D015534:Trans-Activators; D000071230:Transcriptional Regulator ERG; D002110:Caffeine; D013973:Thyrotropin-Releasing Hormone; D016589:Astemizole; D002118:Calcium