NGF, cyclic AMP, and phorbol esters regulate oxytocin receptor gene transcription in SK-N-SH and MCF7 cells

Tracy L. Bale, Daniel Dorsa

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Oxytocin receptor (OTR) gene transcription has predominantly been thought to be regulated by estrogen. However, the continuous presence of receptors in certain brain regions after gonadectomy suggests the existence of alternate mechanisms of regulation. We have cloned and sequenced 4 kb of 5'-flanking DNA of the rat OTR gene and identified an internal segment which was absent in the initial publication of this promoter sequence. Sequence analysis of this segment, as well as of a novel upstream region, revealed the presence of a CRE as well as several other potential regulatory elements, including AP-1, AP-2, AP-3, AP-4 sites, an ERE, and a half-SRE (SRE/2). The effects of phorbol 12-myristate 13-acetate (PMA), forskolin, and NGF treatment on this promoter were tested in transfection experiments in MCF7 and SK-N-SH cells. Transcription of the full-length OTR promoter was induced by forskolin and by the phorbol ester PMA, and a synergistic (17-fold) effect was observed in MCF7 cells treated with both agents. Receptor binding studies using the OTR antagonist 125I-labeled ornithine vasotocin, and Western blot analyses of OTRs in MCF7 cells, showed that PMA and forskolin also increased the density of endogenous human oxytocin receptors. Mutational analyses of the CRE and half-SRE sites in this promoter indicated that these elements function as enhancers and support forskolin and NGF effects, respectively, on transcription. These studies have identified a novel region of the rat OTR promoter containing elements which impart cAMP and/or phorbol ester inducibility of OTR gene transcription. A potential role of the PKA and/or PKC pathways in OTR gene regulation is suggested.

Original languageEnglish (US)
Pages (from-to)130-137
Number of pages8
JournalMolecular Brain Research
Volume53
Issue number1-2
DOIs
StatePublished - Jan 1998
Externally publishedYes

Fingerprint

Oxytocin Receptors
MCF-7 Cells
Phorbol Esters
Nerve Growth Factor
Cyclic AMP
Colforsin
Acetates
Genes
Vasotocin
Ornithine
Transcription Factor AP-1
Transfection
Sequence Analysis
Publications
Estrogens
Western Blotting
DNA
Brain
phorbol-12-myristate

Keywords

  • Forskolin
  • MCF7
  • Oxytocin receptor
  • Phorbol 12-myristate 13-acetate
  • Promoter
  • SK-N-SH
  • Transfection

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

Cite this

NGF, cyclic AMP, and phorbol esters regulate oxytocin receptor gene transcription in SK-N-SH and MCF7 cells. / Bale, Tracy L.; Dorsa, Daniel.

In: Molecular Brain Research, Vol. 53, No. 1-2, 01.1998, p. 130-137.

Research output: Contribution to journalArticle

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