An in vitro screening system for characterizing the cleft palate-inducing potential of chemicals and underlying mechanisms.
An in vitro organ culture system with developing mouse palates was improved to characterize the cleft palate (CP)-inducing potential of chemicals and underlying mechanisms. Palatal explants collected from gestation day 12 mouse fetuses were cultured with various concentrations of teratogens and examined for palatal development after 48 and 72 h of culture to assess effects of the chemicals on growth and/or fusion of palatal shelves. When the explants were exposed to diphenylhydantoin or 5-fluorouracil, palatal growth was inhibited in a concentration-dependent manner at 48 h. Suppression of the expression of proliferative cell nuclear antigen revealed poor cell proliferation. At 72 h, the incidence of explants with CP was significantly increased in the high-dose groups, suggesting that CP induction is mainly attributable to inhibition of palatal growth. By contrast, retinoic acid and hydrocortisone significantly lowered the rates of fused palates at 72 h in all treated groups, while they exhibited no effects on palatal growth at 48 h even at the highest concentration. Because no apoptosis was found in the epithelial cells at the tip of these palates, these chemicals are suggested to inhibit palatal fusion process by preventing apoptosis.
Shimizu
N
N
Toxicology Division I, Institute of Environmental Toxicology, 4321 Uchimoriya-machi, Mitsukaido-shi, Ibaraki 303-0043, Japan.
Aoyama
H
H
Hatakenaka
N
N
Kaneda
M
M
Teramoto
S
S
D000818:Animals; D002455:Cell Division; D002972:Cleft Palate / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D005314:Embryonic and Fetal Development / Q000187:drug effects*; D005260:Female; D005472:Fluorouracil / Q000633:toxicity; D006854:Hydrocortisone / Q000633:toxicity; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009924:Organ Culture Techniques; D010159:Palate / Q000002:abnormalities / Q000187:drug effects* / Q000196:embryology; D010672:Phenytoin / Q000633:toxicity; D013723:Teratogens / Q000633:toxicity*; D014212:Tretinoin / Q000633:toxicity
D013723:Teratogens; D014212:Tretinoin; D010672:Phenytoin; D005472:Fluorouracil; D006854:Hydrocortisone