title: Cocaine- and amphetamine-regulated transcript is the neurotransmitter regulating the action of cholecystokinin and leptin on short-term satiety in rats.
abstract: Vagal CCK-A receptors (CCKARs) and leptin receptors (LRbs) interact synergistically to mediate short-term satiety. Cocaine- and amphetamine-regulated transcript (CART) peptide is expressed by vagal afferent neurons. We sought to demonstrate that this neurotransmitter regulates CCK and leptin actions on short-term satiety. We also examined the signal transduction pathways responsible for mediating the CART release from the nodose ganglia (NG). ELISA studies coupled with gene silencing of NG neurons by RNA interference elucidated intracellular signaling pathways responsible for CCK/leptin-stimulated CART release. Feeding studies followed by gene silencing of CART in NG established the role of CART in mediating short-term satiety. Immunohistochemistry was performed on rat NG neurons to confirm colocalization of CCKARs and LRbs; 63% of these neurons contained CART. Coadministration of CCK-8 and leptin caused a 2.2-fold increase in CART release that was inhibited by CCK-OPE, a low-affinity CCKAR antagonist. Transfection of cultured NG neurons with steroid receptor coactivator (SRC) or phosphatidylinositol 3-kinase (PI3K) small-interfering RNA (siRNA) or STAT3 lentiviral short hairpin RNA inhibited CCK/leptin-stimulated CART release. Silencing the expression of the EGR-1 gene inhibited the CCK/leptin-stimulated CART release but had no effect on CCK/leptin-stimulated neuronal firing. Electroporation of NG with CART siRNA inhibited CCK/leptin stimulated c-Fos expression in rat hypothalamus. Feeding studies following electroporation of the NG with CART or STAT3 siRNA abolished the effects of CCK/leptin on short-term satiety. We conclude that the synergistic interaction of low-affinity vagal CCKARs and LRbs mediates CART release from the NG, and CART is the principal neurotransmitter mediating short-term satiety. CART release from the NG involves interaction between CCK/SRC/PI3K cascades and leptin/JAK2/PI3K/STAT3 signaling pathways.
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan 48109-5362, USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Heldsinger
authors.forename: Yuanxu
authors.initials: Y
authors.lastname: Lu
authors.forename: Shi-Yi
authors.initials: SY
authors.lastname: Zhou
authors.forename: Xiaoyin
authors.initials: X
authors.lastname: Wu
authors.forename: Gintautas
authors.initials: G
authors.lastname: Grabauskas
authors.forename: Il
authors.initials: I
authors.lastname: Song
authors.forename: Chung
authors.initials: C
authors.lastname: Owyang
mesh_terms: D000818:Animals; D015536:Down-Regulation; D018274:Electroporation; D020868:Gene Silencing; D007150:Immunohistochemistry; D020738:Leptin / Q000378:metabolism / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009620:Nodose Ganglion / Q000187:drug effects / Q000378:metabolism; D056918:Nuclear Receptor Coactivators / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D043886:Receptor, Cholecystokinin A / Q000378:metabolism; D054411:Receptors, Leptin / Q000378:metabolism; D012527:Satiation / Q000187:drug effects*; D012844:Sincalide / Q000378:metabolism / Q000494:pharmacology; D009435:Synaptic Transmission / Q000235:genetics; D000098882:Cocaine- and Amphetamine-Regulated Transcript Protein
chemical_list: D020738:Leptin; D009419:Nerve Tissue Proteins; D056918:Nuclear Receptor Coactivators; D034741:RNA, Small Interfering; D043886:Receptor, Cholecystokinin A; D054411:Receptors, Leptin; D000098882:Cocaine- and Amphetamine-Regulated Transcript Protein; D012844:Sincalide