Histamine H1-receptors mediate phosphoinositide hydrolysis in astrocyte-enriched primary cultures

Abstract

Astrocyte-enriched primary cultures of newborn rat brain hemispheres, prelabeled with [3H]inositol, accumulated [3H]inositol phosphate but not [3H]inositol bis- and tris-phosphate, after exposure to histamine for 60 min in the presence of 10 mM LiCl. The response to histamine was not a function of contaminating meningeal fibroblasts since no accumulation of [3H]inositol phosphate was elicited by histamine in meningeal cultures. The stimulation of phosphoinositide hydrolysis by histamine in astrocytes was dose-dependent (EC50 = 1.7 microM, maximal effect = 345% over basal levels) and was mimicked by several H1-receptor agonists. The use of selective receptor antagonists confirmed that the histamine response was the result of activation of H1-receptors. The histamine-induced [3H]inositol phosphate accumulation was completely abolished by omission of Ca2+ from the incubation medium. Astrocyte membranes specifically bound the radiolabeled H1-antagonist, [3H]mepyramine with an affinity (Kd = 5.9 nM) and a density of binding sites (Bmax = 113 fmol/mg protein) similar to rat brain. These results demonstrate the presence of functional histamine H1-receptors in rat brain astrocytes and suggest a role for histamine as a neuromodulator of astrocyte function.