title: Influence of intracellular zinc on cultures of rat cardiac neural crest cells.
abstract: BACKGROUND
  Developmental zinc (Zn) deficiency increases the incidence of heart anomalies in rat fetuses, in regions and structures derived from the outflow tract. Given that the development of the outflow tract requires the presence of cardiac neural crest cells (cNCC), we speculated that Zn deficiency selectively kills cNCC and could lead to heart malformations.
  
  
  METHODS
  Cardiac NCC were isolated from E10.5 rat embryos and cultured in control media (CTRL), media containing 3 μM of the cell permeable metal chelator N, N, N', N'-tetrakis (2-pyridylmethyl) ethylene diamine (TPEN), or in TPEN-treated media supplemented with 3 μM Zn (TPEN + Zn). Cardiac NCC were collected after 6, 8, and 24 h of treatment to assess cell viability, proliferation, and apoptosis.
  
  
  RESULTS
  The addition of TPEN to the culture media reduced free intracellular Zn pools and cell viability as assessed by low ATP production, compared to cells grown in control or Zn-supplemented media. There was an accumulation of reactive oxygen species, a release of mitochondrial cytochrome c into the cytoplasm, and an increased cellular expression of active caspase-3 in TPEN-treated cNCC compared to cNCC cultured in CTRL or TPEN + Zn media.
  
  
  CONCLUSIONS
  Zn deficiency can result in oxidative stress in cNCC, and subsequent decreases in their population and metabolic activity. These data support the concept that Zn deficiency associated developmental heart defects may arise in part as a consequence of altered cNCC metabolism.
authors.affiliation: Department of Nutrition, University of California, Davis, California.
authors.forename: Hsunhui
authors.initials: H
authors.lastname: Yang
authors.forename: Carl L
authors.initials: CL
authors.lastname: Keen
authors.forename: Louise
authors.initials: L
authors.lastname: Lanoue
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003300:Copper / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000378:metabolism; D005029:Ethylenediamines / Q000494:pharmacology; D005260:Female; D042541:Intracellular Space / Q000378:metabolism*; D007501:Iron / Q000378:metabolism; D009206:Myocardium / Q000166:cytology*; D009432:Neural Crest / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D015032:Zinc / Q000172:deficiency* / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D005029:Ethylenediamines; D017382:Reactive Oxygen Species; D003300:Copper; D000255:Adenosine Triphosphate; D045304:Cytochromes c; D007501:Iron; D003576:Electron Transport Complex IV; D053148:Caspase 3; D015032:Zinc; C044387:N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine
keywords: apoptosis; neural crest cells; oxidative damage; zinc