title: Prenatal exposure of ethanol induces increased glutamatergic neuronal differentiation of neural progenitor cells.
abstract: BACKGROUND
  Prenatal ethanol exposure during pregnancy induces a spectrum of mental and physical disorders called fetal alcohol spectrum disorder (FASD). The central nervous system is the main organ influenced by FASD, and neurological symptoms include mental retardation, learning abnormalities, hyperactivity and seizure susceptibility in childhood along with the microcephaly. In this study, we examined whether ethanol exposure adversely affects the proliferation of NPC and de-regulates the normal ratio between glutamatergic and GABAergic neuronal differentiation using primary neural progenitor culture (NPC) and in vivo FASD models.
  
  
  METHODS
  Neural progenitor cells were cultured from E14 embryo brain of Sprague-Dawley rat. Pregnant mice and rats were treated with ethanol (2 or 4 g/kg/day) diluted with normal saline from E7 to E16 for in vivo FASD animal models. Expression level of proteins was investigated by western blot analysis and immunocytochemical assays. MTT was used for cell viability. Proliferative activity of NPCs was identified by BrdU incorporation, immunocytochemistry and FACS analysis.
  
  
  RESULTS
  Reduced proliferation of NPCs by ethanol was demonstrated using BrdU incorporation, immunocytochemistry and FACS analysis. In addition, ethanol induced the imbalance between glutamatergic and GABAergic neuronal differentiation via transient increase in the expression of Pax6, Ngn2 and NeuroD with concomitant decrease in the expression of Mash1. Similar pattern of expression of those transcription factors was observed using an in vivo model of FASD as well as the increased expression of PSD-95 and decreased expression of GAD67.
  
  
  CONCLUSIONS
  These results suggest that ethanol induces hyper-differentiation of glutamatergic neuron through Pax6 pathway, which may underlie the hyper-excitability phenotype such as hyperactivity or seizure susceptibility in FASD patients.
authors.affiliation: Department of Pharmacology, College of Pharmacy, Seoul National University, Seoul, Korea.
authors.forename: Ki Chan
authors.initials: KC
authors.lastname: Kim
authors.forename: Hyo Sang
authors.initials: HS
authors.lastname: Go
authors.forename: Hae Rang
authors.initials: HR
authors.lastname: Bak
authors.forename: Chang Soon
authors.initials: CS
authors.lastname: Choi
authors.forename: Inha
authors.initials: I
authors.lastname: Choi
authors.forename: Pitna
authors.initials: P
authors.lastname: Kim
authors.forename: Seol-Heui
authors.initials: SH
authors.lastname: Han
authors.forename: So Min
authors.initials: SM
authors.lastname: Han
authors.forename: Chan Young
authors.initials: CY
authors.lastname: Shin
authors.forename: Kwang Ho
authors.initials: KH
authors.lastname: Ko
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D000431:Ethanol / Q000494:pharmacology*; D005136:Eye Proteins / Q000378:metabolism; D005260:Female; D063647:Fetal Alcohol Spectrum Disorders / Q000503:physiopathology*; D018698:Glutamic Acid / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D055495:Neurogenesis / Q000187:drug effects; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000378:metabolism; D011247:Pregnancy; D011297:Prenatal Exposure Delayed Effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
chemical_list: D005136:Eye Proteins; D018398:Homeodomain Proteins; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; C000605733:Pax6 protein, rat; D012097:Repressor Proteins; D000431:Ethanol; D018698:Glutamic Acid; D005680:gamma-Aminobutyric Acid