title: Inhibitors of choline uptake and metabolism cause developmental abnormalities in neurulating mouse embryos.
abstract: BACKGROUND
  Choline is an essential nutrient in methylation, acetylcholine and phospholipid biosynthesis, and in cell signaling. The demand by an embryo or fetus for choline may place a pregnant woman and, subsequently, the developing conceptus at risk for choline deficiency.
  
  
  METHODS
  To determine whether a disruption in choline uptake and metabolism results in developmental abnormalities, early somite staged mouse embryos were exposed in vitro to either an inhibitor of choline uptake and metabolism, 2-dimethylaminoethanol (DMAE), or an inhibitor of phosphatidylcholine synthesis, 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH(3)). Cell death following inhibitor exposure was investigated with LysoTracker Red and histology.
  
  
  RESULTS
  Embryos exposed to 250-750 microM DMAE for 26 hr developed craniofacial hypoplasia and open neural tube defects in the forebrain, midbrain, and hindbrain regions. Embryos exposed to 125-275 microM ET-18-OCH(3) exhibited similar defects or expansion of the brain vesicles. ET-18-OCH(3)-affected embryos also had a distended neural tube at the posterior neuropore. Embryonic growth was reduced in embryos treated with either DMAE (375, 500, and 750 microM) or ET-18-OCH(3) (200 and 275 microM). Whole mount staining with LysoTracker Red and histological sections showed increased areas of cell death in embryos treated with 275 microM ET-18-OCH(3) for 6 hr, but there was no evidence of cell death in DMAE-exposed embryos.
  
  
  CONCLUSIONS
  Inhibition of choline uptake and metabolism during neurulation results in growth retardation and developmental defects that affect the neural tube and face.
authors.affiliation: Department of Cell Biology and Anatomy, School of Medicine, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: M C
authors.initials: MC
authors.lastname: Fisher
authors.forename: S H
authors.initials: SH
authors.lastname: Zeisel
authors.forename: M H
authors.initials: MH
authors.lastname: Mar
authors.forename: T W
authors.initials: TW
authors.lastname: Sadler
mesh_terms: D000014:Abnormalities, Drug-Induced / Q000209:etiology*; D000818:Animals; D018726:Anti-Dyskinesia Agents / Q000633:toxicity*; D002794:Choline / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D003642:Deanol / Q000633:toxicity*; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000473:pathology; D005314:Embryonic and Fetal Development / Q000187:drug effects*; D005260:Female; D008297:Male; D051379:Mice; D009436:Neural Tube Defects / Q000139:chemically induced* / Q000196:embryology / Q000473:pathology; D009924:Organ Culture Techniques; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology / Q000633:toxicity; D010742:Phospholipid Ethers / Q000494:pharmacology / Q000633:toxicity; D011247:Pregnancy
chemical_list: D018726:Anti-Dyskinesia Agents; D010726:Phosphodiesterase Inhibitors; D010742:Phospholipid Ethers; C026659:edelfosine; D003642:Deanol; D002794:Choline