title: Prolactin is a major inhibitor of hepatic Leptin A synthesis and secretion: studies utilizing a homologous Leptin A ELISA in the tilapia.
abstract: The present study identifies regulatory interactions between leptin A (LepA) and the pituitary hormone prolactin (PRL). In order to measure tilapia (Oreochromis mossambicus) LepA, an enzyme-linked immunosorbent assay (ELISA) utilizing a rabbit polyclonal antibody specific to tilapia LepA was first developed. The antibody shows strong cross reactivity to recombinant tilapia LepA (rtLepA), and a corresponding 16kDa protein in both tilapia and striped bass plasma, but not to recombinant human leptin (rhLep). The assay has a linear detection range of 0.25-1000nM, with intra- and interassay variability of 9% and 16%, respectively. Plasma LepA levels measured in tilapia ranged from 0.8 to 3.9nM, similar to that found for other vertebrates. Hypophysectomy (Hx) increased circulating LepA and lepa mRNA levels in the liver, the dominant source of hormone production. Adminstration of ovine PRL (oPRL, 5μg/g BW) to Hx fish restored circulating LepA and hepatic lepa mRNA levels to those of control fish. Additionally, oPRL reduced lepa mRNA levels in a dose-dependent fashion in cultured hepatocytes following an 18h incubation. Previous work in our lab indicates that rhLep stimulates PRL release in vitro from tilapia pituitaries. Here, both rtLepA and rhLep (0.5μg/g BW) increased mRNA expression of tilapia prolactin mRNAs (prl1, prl2) in the pituitary in vivo. These results demonstrate that LepA enhances pituitary prolactin synthesis and release, while PRL in turn inhibits hepatic leptin secretion and synthesis in teleosts. We postulate this regulatory interaction may be necessary for mobilizing energy reserves during acute hyperosmotic adaptation.
authors.affiliation: Department of Biological Sciences, North Carolina State University, Raleigh, NC 27695-7617, USA.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Douros
authors.affiliation: Department of Biological Sciences, North Carolina State University, Raleigh, NC 27695-7617, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Baltzegar
authors.affiliation: Department of Biology, Center for Neuroendocrine Studies, University of Massachusetts, Amherst, MA 01003, USA.
authors.forename: Jason P
authors.initials: JP
authors.lastname: Breves
authors.affiliation: Hawaii Institute of Marine Biology, University of Hawaii, Kaneohe, HI 96744, USA.
authors.forename: Darren T
authors.initials: DT
authors.lastname: Lerner
authors.affiliation: Hawaii Institute of Marine Biology, University of Hawaii, Kaneohe, HI 96744, USA.
authors.forename: Andre P
authors.initials: AP
authors.lastname: Seale
authors.affiliation: Hawaii Institute of Marine Biology, University of Hawaii, Kaneohe, HI 96744, USA.
authors.forename: E
authors.initials: E
authors.lastname: Gordon Grau
authors.affiliation: Department of Biological Sciences, North Carolina State University, Raleigh, NC 27695-7617, USA. Electronic address: russell_borski@ncsu.edu.
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Borski
mesh_terms: D000064:Acclimatization; D000818:Animals; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007016:Hypophysectomy; D020738:Leptin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism*; D011388:Prolactin / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017210:Tilapia / Q000254:growth & development / Q000378:metabolism*
chemical_list: D020738:Leptin; D012333:RNA, Messenger; D011994:Recombinant Proteins; D011388:Prolactin
keywords: Enzyme-linked immunosorbant assay; Hepatocytes; Leptin; Pituitary; Prolactin; Teleost fishes