IKir regulation in murine macrophages: whole cell and perforated patch studies.

Abstract

Previous studies have reported that the inwardly rectifying K+ conductance (GKir) in macrophages is modulated by intracellular perfusion with inositol 1,4,5-triphosphate (InsP3), inositol 1,3,4,5-tetrakisphosphate (InsP4), or GTP analogues and by exposing cells to macrophage-specific colony-stimulating factor (CSF) I. This study uses both conventional whole cell and amphotericin B perforated patch studies to investigate GKir modulation in thioglycollate-elicited mouse peritoneal macrophages (MO). Under whole cell recording conditions with 150 mM Cl- in the patch pipette, GKir decreased within 25 min. The GKir decrease was slowed by exchanging glutamate for Cl- as the major anion in the pipette or by adding guanosine 5'-O-(2-thiodiphosphate) (50 nM) or ATP (0.5 mM) to the pipette. Addition of InsP3 or InsP4 to the pipette had no effect on the magnitude of GKir or its rate of decrease but activated an outward current in the voltage range of +60 to +120 mV in 57% of the cells studied. Thus in murine MO GKir may be modulated by G proteins but is unaffected by inositol phosphate metabolites, which have been reported to enhance GKir in phorbol 12-myristate 13-acetate (PMA)-differentiated HL-60 cells. In contrast to whole cell studies, perforated patch recordings of murine MO GKir were stable for > 1 h. Perforated patch studies demonstrated that murine MO also differ from PMA-differentiated HL-60 cells in that CSF I had no effect on GKir. Additionally, arachidonic acid, PMA, and H2O2, agents implicated in macrophage activation, did not modulate GKir. We conclude that GKir regulation in murine MO differs from that reported in PMA-differentiated HL-60 cells and that although our data suggest that GKir is modulated by G protein(s), they differ from the G proteins involved in MO responses to CSF I and the other agents tested.