Two isoforms of prostaglandin E receptor EP3 subtype. Different COOH-terminal domains determine sensitivity to agonist-induced desensitization.
We recently identified two isoforms of mouse prostaglandin (PG) E receptor EP3 subtype, EP3 alpha and EP 3 beta, which are produced by alternative splicing and different only in the carboxyl-terminal domain (Sugimoto, Y., Negishi, M., Hayashi, Y., Namba, Y., T., Honda, A., Watabe, A., Hirata, M., Narumiya, S., and Ichikawa, A. (1993) J. Biol. Chem. 268, 2712-2718). We examined here agonist-induced desensitization of the two isoforms using Chinese hamster ovary cells stably expressing these isoforms. Exposure of the EP3 alpha isoform to PGE2 for 30 min did not change maximal response but increased PGE2 concentration needed to inhibit forskolin-induced cAMP accumulation in the cells. Further exposure of this isoform to PGE2 suppressed the maximal response as well as sensitivity to PGE2 in a time-dependent manner; after 24-h exposure, it elicited only 50% of the maximal response of the control cells. Consistent with these results, short term exposure sequestered the EP3 alpha isoform away from the cell surface and long term incubation decreased the total receptor number in the cells. In contrast, exposure of the EP3 beta isoform to PGE2 did not affect its dose-response curve for PGE2, and no sequestration or decrease in the receptor number was observed in this isoform. Thus, alternative splicing produced the two isoforms with different carboxyl-terminal domains, which are different in sensitivity to agonist-induced desensitization.
Negishi
M
M
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Sugimoto
Y
Y
Irie
A
A
Narumiya
S
S
Ichikawa
A
A
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002850:Chromatography, Gel; D005576:Colforsin / Q000494:pharmacology; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism* / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects; D007700:Kinetics; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D011982:Receptors, Prostaglandin / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D018078:Receptors, Prostaglandin E; D011994:Recombinant Proteins / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection
D046911:Macromolecular Substances; D011982:Receptors, Prostaglandin; D018078:Receptors, Prostaglandin E; D011994:Recombinant Proteins; D005576:Colforsin; D000242:Cyclic AMP; D015232:Dinoprostone