Inducible Cyclic AMP Early Repressor Protein in Rat Pinealocytes: A Highly Sensitive Natural Reporter for Regulated Gene Transcription
- 1Dr. Senckenbergische Anatomie, Institute for Anatomy II, Johann Wolfgang Goethe-University Frankfurt, Frankfurt, Germany (M.P., E.M., H.-W.K., J.H.S.); and 2Department of Obstetrics and Gynecology, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, New Jersey (C.A.M.)
Abstract
Rhythmic activity of arylalkylamine N-acetyltransferase (AANAT) determines melatonin synthesis in rat pineal gland. The transcriptional regulation of AANAT involves the activating and inhibiting transcription factors of the cyclic AMP (cAMP)-signaling pathway, cAMP response element-binding protein and inducible cAMP early repressor (ICER), respectively. Activation of this pathway is centered around norepinephrine, stimulating β1-adrenergic receptors, but various other transmitters can modulate melatonin biosynthesis. To compare the transcriptional impact of norepinephrine with that of other neurotransmitters on melatonin synthesis, we determined ICER protein levels in pinealocytes and, in parallel, hormone secretion. The dose-dependent inductions of ICER protein by norepinephrine, the β1-adrenergic receptor agonist isoproterenol, vasoactive intestinal peptide, pituitary adenylate cyclase-activating polypeptide, and adenosine are correlated to regulatory dynamics in melatonin production. Importantly, ICER protein induction required lower ligand concentrations than the induction of melatonin biosynthesis. Although neuropeptide Y, glutamate, and vasopressin altered norepinephrine-stimulated hormone production without affecting ICER levels, the activation of voltage-gated cation channels increased ICER without affecting hormone synthesis. Sensitivity and versatility of ICER induction in pinealocytes make these neuroendocrine cells a valuable model system in which to study molecular interactions determining a regulated gene expression.
Footnotes
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Send reprint requests to: Dr. Jörg H. Stehle, Dr. Senckenbergische Anatomie, Anatomisches Institut II, Hs 26, Johann Wolfgang Goethe-Universität Frankfurt, Theodor-Stern-Kai7, 60590 Frankfurt, Germany. E-mail:stehle{at}em.uni-frankfurt.de
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This work was supported by grants from Deutsche Forschungsgemeinschaft (H.-W.K., J.H.S.) and August-Scheidel-Stiftung (J.H.S.).
- Abbreviations:
- AANAT
- arylalkylamineN-acetyltransferase
- AC
- adenylate cyclase
- cAMP
- cyclic AMP
- ACh
- acetylcholine
- AVP
- arginine-vasopressin
- CREB
- cyclic AMP response element-binding protein
- ICER
- inducible cyclic AMP early repressor
- ISO
- isoproterenol
- IR
- immunoreaction
- NE
- norepinephrine
- NECA
- 5′-N-ethylcarboxy-amidoadenosine
- NGF
- nerve growth factor
- NPY
- neuropeptide Y
- PACAP
- pituitary adenylate cyclase-activating polypeptide
- pCREB
- phosphorylated cyclic AMP response element-binding protein
- PHE
- phenylephrine
- PRAZO
- prazosin
- PROP
- propranolol
- VP4-9
- arginine-vasopressin fragment 4-9
- TF
- transcription factor
- VIP
- vasoactive intestinal peptide
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- Received March 11, 1999.
- Accepted April 29, 1999.
- The American Society for Pharmacology and Experimental Therapeutics
