title: Neural differentiation from human embryonic stem cells as a tool to study early brain development and the neuroteratogenic effects of ethanol.
abstract: The in vitro generation of neural cells from human embryonic stem cells is a powerful tool to acquire better knowledge of the cellular and molecular events involved in early human neural and brain development under physiological and pathological conditions. Prenatal alcohol exposure can induce important anomalies in the developing brain, the embryogenesis being an important critical period for the craniofacial defects and mental disabilities associated with fetal alcohol syndrome. Here, we report the generation of neural progenitors (NPs) from human embryonic stem cells. Neuroepithelial progenitors display the morphological and functional characteristics of their embryonic counterparts and the proper timing of neurons and glia cells generation. Immunocytochemical and real time (RT)-polymerase chain reaction analyses reveal that cells appeared as clusters during neuroepithelial cell proliferation and that the genes associated with the neuroectodermal (Pax-6) and the endodermic (α-fetoprotein) lineages decreased in parallel to the upregulation of the genes of NPs (nestin and Tuj1), followed by their differentiation into neurons (MAP-2+, GABA+), oligodendrocytes [galactocerebroside (GalC+)], and astrocytes (GFAP+). We further demonstrate, for the first time, that human NPs express the endocannabinoid receptors (CB1 and CB2) and the enzymes involved in endocannabinoids synthesis (NAPE-PLD) and degradation (FAAH). Using this in vitro culture, we demonstrate that ethanol exposure impairs NPs survival, affects the differentiation of NPs into neurons and astrocytes, disrupts the actin cytoskeleton, and affects the expression of different genes associated with neural differentiation. The results provide new insights into the effects of ethanol on human embryogenesis and neuroprogenitors and offer an opportunity to delineate potential therapeutic strategies to restore early ethanol-induced brain damage.
authors.affiliation: Cell Pathology Laboratory, Centro de Investigación Príncipe Felipe, Valencia, Spain.
authors.forename: Raquel
authors.initials: R
authors.lastname: Taléns-Visconti
authors.forename: Irene
authors.initials: I
authors.lastname: Sanchez-Vera
authors.forename: Jelena
authors.initials: J
authors.lastname: Kostic
authors.forename: Maria Amparo
authors.initials: MA
authors.lastname: Perez-Arago
authors.forename: Slaven
authors.initials: S
authors.lastname: Erceg
authors.forename: Miodrag
authors.initials: M
authors.lastname: Stojkovic
authors.forename: Consuelo
authors.initials: C
authors.lastname: Guerri
mesh_terms: D001921:Brain / Q000187:drug effects* / Q000254:growth & development; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects*; D000431:Ethanol / Q000633:toxicity*; D020869:Gene Expression Profiling; D006801:Humans; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D013723:Teratogens / Q000633:toxicity*
chemical_list: D013723:Teratogens; D000431:Ethanol