Prostaglandin D2(PGD2) has marked inhibitory effects on the canine proximal colonic epithelium set up in Ussing chambers. These effects involved a receptor that is pharmacologically distinct from the classical DP, presumably the recently identified CRTH2/DP2 variety. The mechanism underlying these effects was studied using 13,14-dihydro-15-keto-PGD2(DK-PGD2), a stable metabolite of the parent prostanoid. The metabolite quickly reversed short circuit currents (Isc) stimulated by diverse agonists. Greater inhibitory effects were seen with stimulants such as carbachol and cyclopiazonic acid (CPA) rather than with forskolin or protein kinase A activators. Since the same stimulants were differentially affected by removal and replacement of serosal Ca2+, we tested the possibility that the prostanoid inhibited basolateral Ca2+entry. In the absence of serosal Ca2+, tissues primed with CPA demonstrated concentration-dependent increases in Iscto cumulative additions of Ca2+or Sr2+, though the former was more potent. Clremoval and pretreatment with bumetanide virtually abolished responses, suggesting that the increase in Iscreflected Ca2+dependent Clsecretion. Though responses were insensitive to the L-type channel antagonist, verapamil, a marked inhibition was seen in the presence of metal cations (Gd3+, Cd2+, and La3+). Pretreatment with DK-PGD2inhibited responses to Ca2+in CPA-primed tissues. Thus, basolateral Ca2+entry via store-operated Ca2+channels may be the locus for the inhibitory effects of PGD2in this tissue. These results could indicate a potential transduction mechanism for the novel DP receptor variously called CRTH2 or DP2.Key words: store-operated Ca2+channels, capacitative, arachidonic acid metabolites, epithelia.