Molecular cloning and expression of multiple isoforms of human prostaglandin E receptor EP3 subtype generated by alternative messenger RNA splicing: Multiple second messenger systems and tissue-specific distributions

M. Kotani, I. Tanaka, Y. Ogawa, T. Usui, K. Mori, A. Ichikawa, S. Narumiya, T. Yoshimi, K. Nakao

Research output: Contribution to journalArticle

118 Citations (Scopus)

Abstract

Five distinct cDNA clones encoding four different isoforms of human prostaglandin (PG) E receptor EP3 subtype were isolated from a human kidney cDNA library. Two cDNA clones differed only in their 3'-untranslated regions. The four isoforms, tentatively named EP(3-I), EP(3-II), EP(3-III), and EP(3- IV), which were generated by alternative mRNA splicing, had identical amino acid sequences except for their different carboxyl-terminal tails. Transfection experiments revealed that all the four isoforms show high binding affinities to PGE2, PGE1, and M and B28767, an EP3-specific agonist, whereas their downstream signaling pathways are divergent. M and B28767 increased cAMP concentrations in cells expressing EP(3-II) and EP(3- IV), whereas it inhibited forskolin-induced cAMP accumulations in cells expressing all EP3 isoforms. M and B28767 also stimulated phosphoinositide turnover in cells expressing EP(3-I) and EP(3-II). Northern blot analysis revealed that the EP3 gene is expressed in a wide variety of human tissues. The human EP3 mRNA was present most abundantly in the kidney, pancreas, and uterus. A substantial expression was also detected in the heart, liver, skeletal muscle, small intestine, colon, prostate, ovary, and testis. Furthermore, reverse transcription-polymerase chain reaction analysis demonstrated tissue-specific expressions of the five different EP3 mRNA species. The present study suggests the presence of the multiple systems of PGE2/EP3 isoforms and leads to the better understanding of its physiological and pathophysiological implications in humans.

Original languageEnglish
Pages (from-to)869-879
Number of pages11
JournalMolecular Pharmacology
Volume48
Issue number5
Publication statusPublished - Jan 1 1995
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

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