Scientific Results

A Tph2GFP Reporter Stem Cell Line To Model in Vitro and in Vivo Serotonergic Neuron Development and Function.

Year: 2017

Authors: Pacini G., Marino A., Migliarini S., Brilli E., Pelosi B., Maddaloni G., Pratelli M., Pellegrino M., Ferrari A., Pasqualetti M.

Autors Affiliation: Department of Biology, Unit of Cell and Developmental Biology, University of Pisa , 56127 Pisa, Italy.
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.
ETH Zurich , Laboratory of Thermodynamics in Emerging Technologies, 8092 Zurich, Switzerland.
Center for Neuroscience and Cognitive Systems, Istituto Italiano di Tecnologia , 38068 Rovereto (TN), Italy.
Neuroscience Institute, National Research Council (CNR) , 56124 Pisa, Italy.

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 Tph2(GFP), in which the vital reporter GFP replaces the tryptophan hydroxylase 2 (Tph2) gene. Tph2GFP mES 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 Tph2GFP cells are able to terminally differentiate, integrate, and innervate the host brain when grafted in vivo. On the whole, the present study introduces the Tph2GFP mES 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.

Journal/Review: ACS CHEMICAL NEUROSCIENCE

Volume: 8 (5)      Pages from: 1043  to: 1052

KeyWords: 5-HT; Embryonic stem cells; Serotonergic neurons
DOI: 10.1021/acschemneuro.6b00403

Citations: 4
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English