Functional characterization of rhesus embryonic stem cell-derived serotonin neurons

Yukari Tokuyama, Susan Ingram, Joy S. Woodward, Cynthia Bethea

    Research output: Contribution to journalArticle

    9 Citations (Scopus)

    Abstract

    Optimal function of the serotonin system is essential for mental health and its role in psychopathologies is undisputed. Enhancing the ability to study primate serotonin neurons in culture would facilitate understanding of intracellular signaling pathways that mediate the action of drugs and other epigenetic or developmental factors impacting human mental health. We were the first group to report differentiation of the non-human primate rhesus monkey embryonic stem cell (ESC) line 366.4 into cultures of serotonin neurons. In this study, we optimized yield and obtained functional characteristics of the derived serotonin neurons. Sequential treatments of ESC 366.4 during expansion stage with fibroblast growth factor 4 and sonic hedgehog markedly increased the yield of serotonin neurons. These serotonin neurons propagated action potentials and expressed GABA receptors. Also, for the first time we demonstrate that these ESC-derived serotonin neurons exhibit functional high-affinity transporter sites, as well as high-affinity 5HT1A binding sites, which are essential targets of common psychoactive drugs. Finally, to test the generality of this method, we utilized another rhesus ESC line, ORMES-22, which efficiently differentiated into serotonin neurons. Together, these findings demonstrate the feasibility of our protocol to direct different primate ESC lines to serotonin neurons with physiological characteristics, which makes them a useful in vitro model system.

    Original languageEnglish (US)
    Pages (from-to)649-657
    Number of pages9
    JournalExperimental Biology and Medicine
    Volume235
    Issue number5
    DOIs
    StatePublished - May 2010

    Fingerprint

    Embryonic Stem Cells
    Stem cells
    Neurons
    Serotonin
    Primates
    Cell Line
    Mental Health
    Fibroblast Growth Factor 4
    Health
    Aptitude
    GABA Receptors
    Hedgehogs
    Psychotropic Drugs
    Human engineering
    Macaca mulatta
    Psychopathology
    Cell culture
    Epigenomics
    Action Potentials
    Binding Sites

    Keywords

    • 5HT autoreceptor
    • Differentiation
    • Embryonic stem cells
    • Rhesus macaque
    • Serotonin
    • Serotonin reuptake transporter

    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)
    • Medicine(all)

    Cite this

    Functional characterization of rhesus embryonic stem cell-derived serotonin neurons. / Tokuyama, Yukari; Ingram, Susan; Woodward, Joy S.; Bethea, Cynthia.

    In: Experimental Biology and Medicine, Vol. 235, No. 5, 05.2010, p. 649-657.

    Research output: Contribution to journalArticle

    @article{f393753c58f343d2b436f653173c9949,
    title = "Functional characterization of rhesus embryonic stem cell-derived serotonin neurons",
    abstract = "Optimal function of the serotonin system is essential for mental health and its role in psychopathologies is undisputed. Enhancing the ability to study primate serotonin neurons in culture would facilitate understanding of intracellular signaling pathways that mediate the action of drugs and other epigenetic or developmental factors impacting human mental health. We were the first group to report differentiation of the non-human primate rhesus monkey embryonic stem cell (ESC) line 366.4 into cultures of serotonin neurons. In this study, we optimized yield and obtained functional characteristics of the derived serotonin neurons. Sequential treatments of ESC 366.4 during expansion stage with fibroblast growth factor 4 and sonic hedgehog markedly increased the yield of serotonin neurons. These serotonin neurons propagated action potentials and expressed GABA receptors. Also, for the first time we demonstrate that these ESC-derived serotonin neurons exhibit functional high-affinity transporter sites, as well as high-affinity 5HT1A binding sites, which are essential targets of common psychoactive drugs. Finally, to test the generality of this method, we utilized another rhesus ESC line, ORMES-22, which efficiently differentiated into serotonin neurons. Together, these findings demonstrate the feasibility of our protocol to direct different primate ESC lines to serotonin neurons with physiological characteristics, which makes them a useful in vitro model system.",
    keywords = "5HT autoreceptor, Differentiation, Embryonic stem cells, Rhesus macaque, Serotonin, Serotonin reuptake transporter",
    author = "Yukari Tokuyama and Susan Ingram and Woodward, {Joy S.} and Cynthia Bethea",
    year = "2010",
    month = "5",
    doi = "10.1258/ebm.2010.009307",
    language = "English (US)",
    volume = "235",
    pages = "649--657",
    journal = "Experimental Biology and Medicine",
    issn = "1535-3702",
    publisher = "SAGE Publications Ltd",
    number = "5",

    }

    TY - JOUR

    T1 - Functional characterization of rhesus embryonic stem cell-derived serotonin neurons

    AU - Tokuyama, Yukari

    AU - Ingram, Susan

    AU - Woodward, Joy S.

    AU - Bethea, Cynthia

    PY - 2010/5

    Y1 - 2010/5

    N2 - Optimal function of the serotonin system is essential for mental health and its role in psychopathologies is undisputed. Enhancing the ability to study primate serotonin neurons in culture would facilitate understanding of intracellular signaling pathways that mediate the action of drugs and other epigenetic or developmental factors impacting human mental health. We were the first group to report differentiation of the non-human primate rhesus monkey embryonic stem cell (ESC) line 366.4 into cultures of serotonin neurons. In this study, we optimized yield and obtained functional characteristics of the derived serotonin neurons. Sequential treatments of ESC 366.4 during expansion stage with fibroblast growth factor 4 and sonic hedgehog markedly increased the yield of serotonin neurons. These serotonin neurons propagated action potentials and expressed GABA receptors. Also, for the first time we demonstrate that these ESC-derived serotonin neurons exhibit functional high-affinity transporter sites, as well as high-affinity 5HT1A binding sites, which are essential targets of common psychoactive drugs. Finally, to test the generality of this method, we utilized another rhesus ESC line, ORMES-22, which efficiently differentiated into serotonin neurons. Together, these findings demonstrate the feasibility of our protocol to direct different primate ESC lines to serotonin neurons with physiological characteristics, which makes them a useful in vitro model system.

    AB - Optimal function of the serotonin system is essential for mental health and its role in psychopathologies is undisputed. Enhancing the ability to study primate serotonin neurons in culture would facilitate understanding of intracellular signaling pathways that mediate the action of drugs and other epigenetic or developmental factors impacting human mental health. We were the first group to report differentiation of the non-human primate rhesus monkey embryonic stem cell (ESC) line 366.4 into cultures of serotonin neurons. In this study, we optimized yield and obtained functional characteristics of the derived serotonin neurons. Sequential treatments of ESC 366.4 during expansion stage with fibroblast growth factor 4 and sonic hedgehog markedly increased the yield of serotonin neurons. These serotonin neurons propagated action potentials and expressed GABA receptors. Also, for the first time we demonstrate that these ESC-derived serotonin neurons exhibit functional high-affinity transporter sites, as well as high-affinity 5HT1A binding sites, which are essential targets of common psychoactive drugs. Finally, to test the generality of this method, we utilized another rhesus ESC line, ORMES-22, which efficiently differentiated into serotonin neurons. Together, these findings demonstrate the feasibility of our protocol to direct different primate ESC lines to serotonin neurons with physiological characteristics, which makes them a useful in vitro model system.

    KW - 5HT autoreceptor

    KW - Differentiation

    KW - Embryonic stem cells

    KW - Rhesus macaque

    KW - Serotonin

    KW - Serotonin reuptake transporter

    UR - http://www.scopus.com/inward/record.url?scp=77952579474&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=77952579474&partnerID=8YFLogxK

    U2 - 10.1258/ebm.2010.009307

    DO - 10.1258/ebm.2010.009307

    M3 - Article

    VL - 235

    SP - 649

    EP - 657

    JO - Experimental Biology and Medicine

    JF - Experimental Biology and Medicine

    SN - 1535-3702

    IS - 5

    ER -