title: Catecholamine secretory vesicle stimulus-transcription coupling in vivo. Demonstration by a novel transgenic promoter/photoprotein reporter and inhibition of secretion and transcription by the chromogranin A fragment catestatin.
abstract: Stimulation of chromaffin cell secretion in vitro triggers not only secretion but also resynthesis of just released catecholamines and chromogranin A, the precursor of the catecholamine release-inhibitory, nicotinic cholinergic antagonist peptide catestatin. Does stimulus-transcription coupling occur in vivo? And does catestatin antagonize secretion and transcription in vivo? To answer these questions, we employed a novel mouse strain harboring a chromogranin A promoter/firefly luciferase reporter transgene. Tissue-specific expression of the reporter was established by both luminescence and reverse transcription-PCR. Secretion and transcription in vivo were triggered by either direct nicotinic stimulation or vesicular transmitter depletion. Nicotinic blockade in vivo was attempted with either the classical antagonist chlorisondamine or the novel antagonist catestatin. Luciferase reporter expression was exquisitely sensitive over a large dynamic range, was specific for the transgenic animals, and paralleled typical neuroendocrine distribution of endogenous chromogranin A. Adrenal ontogeny revealed a rise of embryonic transgene expression until embryonal day 18, with an abrupt postnatal decline. Direct nicotinic stimulation of chromaffin cells caused catecholamine release and transgene transcription, each of which was nearly completely blocked by chlorisondamine. Similar adrenal results were obtained during vesicular catecholamine depletion. Both secretion and transcription were substantially blocked in the adrenal gland by catestatin. In brain and sympathetic nerve, stimulation of transcription was more modest, and reserpine responses were only incompletely blocked by chlorisondamine or catestatin, perhaps because of limited blood-brain barrier penetration by these cationic antagonists. Thus, nicotinic cholinergic stimulus-transcription coupling occurs in vivo and can be provoked either directly or indirectly (by vesicular transmitter depletion). Such coupling triggers the biosynthesis of chromogranin A, the precursor of catestatin. Catestatin itself blocks stimulation of both secretion and transcription in vivo. Thus, chromogranin A and its catestatin fragment may lie at the nexus of nicotinic cholinergic signaling in vivo.
authors.affiliation: Department of Medicine and Center for Molecular Genetics, University of California, and Veterans Affairs San Diego Healthcare System, San Diego, California 92161, USA.
authors.forename: Sushil K
authors.initials: SK
authors.lastname: Mahata
authors.forename: Nitish R
authors.initials: NR
authors.lastname: Mahapatra
authors.forename: Manjula
authors.initials: M
authors.lastname: Mahata
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Wang
authors.forename: Brian P
authors.initials: BP
authors.lastname: Kennedy
authors.forename: Michael G
authors.initials: MG
authors.lastname: Ziegler
authors.forename: Daniel T
authors.initials: DT
authors.lastname: O'Connor
mesh_terms: D000311:Adrenal Glands / Q000196:embryology / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000196:embryology / Q000378:metabolism; D002395:Catecholamines / Q000378:metabolism*; D053379:Chromogranin A; D002864:Chromogranins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D005260:Female; D008156:Luciferases / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011978:Receptors, Nicotinic / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
chemical_list: D002395:Catecholamines; D053379:Chromogranin A; D002864:Chromogranins; D017931:DNA Primers; D010446:Peptide Fragments; D011978:Receptors, Nicotinic; C109084:chromogranin A (344-364); C506155:chromogranin A, mouse; D008156:Luciferases