Livers from connexin32 (Cx32) deficient mice have been shown to be defective in hormonally induced glucose mobilization. In order to determine whether this effect is due to decreased diffusion of the second messenger inositol 1,4,5-trisphosphate (IP(3)) between hepatocytes, we injected iontophoretically different amounts of IP(3) in Fura-2 loaded hepatocyte doublets (i.e. cell pairs) from wild type or Cx32 deficient mice. Whereas 84% of wild type hepatocytes showed an intercellular Ca(2+) wave spreading from the injected cell to the neighboring cell, only 25% of Cx32 deficient hepatocyte doublets did so. The amount of IP(3) necessary to induce an intercellular Ca(2+) wave in Cx32 deficient hepatocyte doublets was estimated to be about 25-fold higher than in wild type doublets. This confirms the notion that the low hormonally or electrically induced glucose mobilization found in Cx32 deficient livers relative to wild type livers is due to largely hindered diffusion of IP(3) between Cx32 deficient hepatocytes.