Expression profile of differentiating serotonin neurons derived from rhesus embryonic stem cells and comparison to adult serotonin neurons

Cynthia Bethea, Arubala Reddy, Darlene Pedersen, Yukari Tokuyama

    Research output: Contribution to journalArticle

    14 Citations (Scopus)

    Abstract

    The rhesus monkey embryonic stem cell line 366.4 differentiates into serotonin neurons. We examined the genetic cascade during differentiation and compared ESC-derived serotonin neurons to adult monkey serotonin neurons. RNA was extracted from ESC colonies, embryoid bodies (EBs), neurospheres in selection (N1) and proliferation stages (N2), differentiated serotonin neurons (N3) and from laser captured (LC) serotonin neurons of spayed female macaques treated with placebo, estrogen (E), progesterone (P) or E + P. The RNA was labeled and hybridized to Rhesus Monkey Affymetrix Gene Chips (n = 1 per stage and 2 per animal treatment). Gene expression was examined with GeneSifter software. 545 genes that were related to developmental processes showed a threefold or greater change between stages. TGFb, Wnt, VEGF and Hedgehog signaling pathways showed the highest percent of probe set changes during differentiation. Genes in the categories (a) homeobox binding and transcription factors, (b) growth factors and receptors, (c) brain and neural specific factors and (d) serotonin specific factors are reported. Pivotal genes were confirmed with quantitative RT-PCR. In the serotonin developmental cascade, FGFR2 was robustly expressed at each stage. GATA3 was robustly expressed in EBs. Sonic hedgehog (Shh), PTCH (Shh-R) and Fev1 transcription factor expression coincided with the induction of serotonin specific marker genes during N1-selection. A majority of the examined genes were expressed in adult serotonin neurons. However, in the ESC-derived neurons, there was significant over-representation of probe sets related to cell cycle, axon guidance & dorso-ventral axis formation. This analysis suggests that the 366.4 cell line possesses cues for serotonin differentiation at early stages of differentiation, but that ESC-derived serotonin neurons are still immature.

    Original languageEnglish (US)
    Pages (from-to)94-108
    Number of pages15
    JournalGene Expression Patterns
    Volume9
    Issue number2
    DOIs
    StatePublished - Feb 2009

    Fingerprint

    Embryonic Stem Cells
    Serotonin
    Neurons
    Hedgehogs
    Embryoid Bodies
    Genes
    Macaca mulatta
    Transcription Factors
    RNA
    Cell Line
    Growth Factor Receptors
    Homeobox Genes
    Macaca
    Oligonucleotide Array Sequence Analysis
    Vascular Endothelial Growth Factor A
    Haplorhini
    Cues
    Progesterone
    Cell Cycle
    Estrogens

    Keywords

    • Affymetrix array
    • Development
    • Dorsal raphe
    • Embryonic stem cells
    • Gene expression
    • Macaques
    • Serotonin

    ASJC Scopus subject areas

    • Genetics
    • Molecular Biology
    • Developmental Biology

    Cite this

    Expression profile of differentiating serotonin neurons derived from rhesus embryonic stem cells and comparison to adult serotonin neurons. / Bethea, Cynthia; Reddy, Arubala; Pedersen, Darlene; Tokuyama, Yukari.

    In: Gene Expression Patterns, Vol. 9, No. 2, 02.2009, p. 94-108.

    Research output: Contribution to journalArticle

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