Evidence that the gap junction protein connexin-43 is the ATP-induced pore of mouse macrophages

Abstract

Extracellular ATP4- opens pores in the plasma membrane of mouse macrophages and the J774 macrophage-like cell line that allow molecules as large as fura-2 (831 daltons) to enter the cytoplasmic matrix of the cells. The functional similarity of the ATP-induced pores to gap junctions led us to examine whether these pores were related to members of the connexin family of gap junction proteins. Under conditions of high stringency, RNA isolated from J774 cells hybridized with cDNA for connexin-43 but not with cDNA for connexin-32, -26, or -46. RNA isolated from several variant J774 cell lines that do not permeabilize in response to extracellular ATP (ATPR cells) did not hybridize with connexin-43 cDNA. Immunoblots demonstrated that J774 cells, but not the variant ATPR B2 cell line, expressed connexin-43 protein. These studies demonstrate that mouse macrophages express the connexin-43 gap junction mRNA and protein and strongly suggest that in these cells connexin-43 forms "half-gap junctions" in response to extracellular ATP4-.