title: Human serotonin transporter expression during megakaryocytic differentiation of MEG-01 cells.
abstract: The serotonin (5-HT) transporter (SERT) has been found altered in platelets of patients with genetically complex disorders, including mood-anxiety, pain and eating disorders. In this study, we used cell cultures of platelet precursors as models of investigation on mechanisms of SERT regulation: SERT expression was appraised during megakaryocytic differentiation of human megakaryoblastic MEG-01 cells. Cells were cultured for 8 days with 10(-7)M 4-beta-12-tetradecanoylphorbol-13-acetate (beta-TPA) in the presence of 10% fetal bovine serum (FBS) and SERT was assessed by real time PCR, immunofluorescence microscopy, Western blot and [(3)H]5-HT re-uptake. Results revealed that SERT is present in control-untreated MEG-01 cells. beta-TPA-differentiating MEG-01 cells showed a redistribution of SERT fluorescence, diffuse to cell bodies and blebs along with a 3-fold SERT mRNA increase and a moderate raise in SERT protein (1.5/1.4-fold) by immunoblot and re-uptake assays. In summary, we have shown herein that control megakaryoblasts express the SERT protein. SERT is modulated by differentiation events, implying that SERT density in platelets is under the control of megakaryocytopoiesis stages. Differentiation of MEG-01 cells can provide considerable insight into interactions between SERT genetics, transmitter-hormonal/homeostatic mechanisms and signaling pathways.
authors.affiliation: Department of Psychiatry, Neurobiology, Pharmacology and Biotechnology, University of Pisa, Pisa, Italy. ggino@farm.unipi.it
authors.forename: Gino
authors.initials: G
authors.lastname: Giannaccini
authors.forename: Laura
authors.initials: L
authors.lastname: Betti
authors.forename: Lionella
authors.initials: L
authors.lastname: Palego
authors.forename: Lara
authors.initials: L
authors.lastname: Schmid
authors.forename: Laura
authors.initials: L
authors.lastname: Fabbrini
authors.forename: Caterina
authors.initials: C
authors.lastname: Pelosini
authors.forename: Claudia
authors.initials: C
authors.lastname: Gargini
authors.forename: Ylenia
authors.initials: Y
authors.lastname: Da Valle
authors.forename: Mario
authors.initials: M
authors.lastname: Lanza
authors.forename: Alessandro
authors.initials: A
authors.lastname: Marsili
authors.forename: Margherita
authors.initials: M
authors.lastname: Maffei
authors.forename: Ferruccio
authors.initials: F
authors.lastname: Santini
authors.forename: Paolo
authors.initials: P
authors.lastname: Vitti
authors.forename: Aldo
authors.initials: A
authors.lastname: Pinchera
authors.forename: Antonio
authors.initials: A
authors.lastname: Lucacchini
mesh_terms: D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics*; D012701:Serotonin / Q000378:metabolism; D050486:Serotonin Plasma Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014908:Triticum
chemical_list: D012333:RNA, Messenger; D050486:Serotonin Plasma Membrane Transport Proteins; D012701:Serotonin; D013755:Tetradecanoylphorbol Acetate