title: A Tph2GFPReporter Stem Cell Line To Model in Vitro and in Vivo Serotonergic Neuron Development and Function.
abstract: Modeling biological systems in vitro has contributed to clarification of complex mechanisms in simplified and controlled experimental conditions. Mouse embryonic stem (mES) cells can be successfully differentiated toward specific neuronal cell fates, thus representing an attractive tool to dissect, in vitro, mechanisms that underlie complex neuronal features. In this study, we generated and characterized a reporter mES cell line, called Tph2GFP, in which the vital reporter GFP replaces the tryptophan hydroxylase 2 (Tph2) gene. Tph2GFPmES cells selectively express GFP upon in vitro differentiation toward the serotonergic fate, they synthesize serotonin, possess excitable membranes, and show the typical morphological, morphometrical, and molecular features of in vivo serotonergic neurons. Thanks to the vital reporter GFP, we highlighted by time-lapse video microscopy several dynamic processes such as cell migration and axonal outgrowth in living cultures. Finally, we demonstrated that predifferentiated Tph2GFPcells are able to terminally differentiate, integrate, and innervate the host brain when grafted in vivo. On the whole, the present study introduces the Tph2GFPmES cell line as a useful tool allowing accurate developmental and dynamic studies and representing a reliable platform for the study of serotonergic neurons in health and disease.
authors.affiliation: Department of Biology, Unit of Cell and Developmental Biology, University of Pisa , 56127 Pisa, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Pacini
authors.affiliation: Department of Biology, Unit of Cell and Developmental Biology, University of Pisa , 56127 Pisa, Italy.
authors.forename: Attilio
authors.initials: A
authors.lastname: Marino
authors.affiliation: Department of Biology, Unit of Cell and Developmental Biology, University of Pisa , 56127 Pisa, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Migliarini
authors.affiliation: Department of Biology, Unit of Cell and Developmental Biology, University of Pisa , 56127 Pisa, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Brilli
authors.affiliation: Department of Biology, Unit of Cell and Developmental Biology, University of Pisa , 56127 Pisa, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Pelosi
authors.affiliation: Department of Biology, Unit of Cell and Developmental Biology, University of Pisa , 56127 Pisa, Italy.
authors.forename: Giacomo
authors.initials: G
authors.lastname: Maddaloni
authors.affiliation: Department of Biology, Unit of Cell and Developmental Biology, University of Pisa , 56127 Pisa, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Pratelli
authors.affiliation: Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, University of Pisa , 56126 Pisa, Italy.|Istituto Nazionale di Ottica, National Research Council (CNR) , 56124 Pisa, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Pellegrino
authors.affiliation: ETH Zurich , Laboratory of Thermodynamics in Emerging Technologies, 8092 Zurich, Switzerland.
authors.forename: Aldo
authors.initials: A
authors.lastname: Ferrari
authors.affiliation: Department of Biology, Unit of Cell and Developmental Biology, University of Pisa , 56127 Pisa, Italy.|Center for Neuroscience and Cognitive Systems, Istituto Italiano di Tecnologia , 38068 Rovereto (TN), Italy.|Neuroscience Institute, National Research Council (CNR) , 56124 Pisa, Italy.
authors.forename: Massimo
authors.identifier: 0000-0002-0844-8139
authors.initials: M
authors.lastname: Pasqualetti
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D059326:Serotonergic Neurons / Q000166:cytology* / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism*; D014365:Tryptophan Hydroxylase / Q000235:genetics / Q000378:metabolism*
chemical_list: D049452:Green Fluorescent Proteins; D012701:Serotonin; D014365:Tryptophan Hydroxylase
keywords: 5-HT; Embryonic stem cells; in vitro differentiation; in vivo grafting; serotonergic neurons; vital serotonergic differentiation sensor