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, Yoshihiro Ogawa, T. Usui, K. Mori, A. Ichikawa, S. Narumiya, T. Yoshimi, K. Nakao

Research output: Contribution to journalArticle

116 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

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Receptors, Prostaglandin E, EP3 Subtype
Alternative Splicing
Molecular Cloning
Second Messenger Systems
Tissue Distribution
Protein Isoforms
Messenger RNA
Complementary DNA
Clone Cells
Kidney
Alprostadil
3' Untranslated Regions
Colforsin
Phosphatidylinositols
Gene Library
Dinoprostone
Northern Blotting
Reverse Transcription
Small Intestine
Uterus

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

Cite this

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

In: Molecular Pharmacology, Vol. 48, No. 5, 01.01.1995, p. 869-879.

Research output: Contribution to journalArticle

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