Incorporation of 5-bromo-2'-deoxyuridine into mesenchymal limb-bud cells destined to die: relationship to polydactyly induction in rats.
The thymidine analogue, 5-bromo-2'-deoxyuridine (BUdR), given at the proper dose and time to pregnant rats produces preaxial hindlimb polydactyly in a high proportion of near term foetuses. The lack of physiological cell death in an area of preaxial mesenchyme known as the foyer primaire preaxial (fpp) is thought to be important in the pathogenesis of this defect. This study addresses the question of whether BUdR's well-known antidifferentiative effects, which are due in some way to drug incorporation into DNA, are operative in this in vivo system. The dose and temporal response of BUdR for the induction of preaxial polydactyly inversely parallels the frequency of embryonic hindlimbs with an fpp. Incorporation of BUdR into degenerative fragments within the fpp of these treated limbs is demonstrated with indirect immunofluorescence using an antibody to bromouridine. Hindlimbs exposed to a threshold dose of BUdR at the optimal time for producing polydactyly have incorporated the drug into degenerative fragments within the fpp. This suggests that a higher, teratogenic dose of BUdR might likewise be incorporated. The resulting higher level of incorporation presumably alters the normal course of terminal differentiation for these cells originally destined to die. Teratogenic doses of BUdR injected at later than the optimal time are also incorporated into dead cell fragments within the fpp, suggesting that presumptive dead cells have additional rounds of DNA synthesis which are BUdR-insensitive. Approximately 12 h prior to overt death presumptive fpp cells no longer incorporate the drug. Results reported support the hypothesis that incorporation of teratogenic levels of BUdR prevent cell death in the fpp. The extra cells are thought to contribute directly or indirectly to the added digit. Contrary to other views, it is suggested that BUdR-induced teratogenesis can be a result of the drug's antidifferentiative effects on specific, 'sensitive', populations of cells.
D000014:Abnormalities, Drug-Induced; D000818:Animals; D001973:Bromodeoxyuridine / Q000633:toxicity*; D002454:Cell Differentiation; D002470:Cell Survival; D004247:DNA / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D005121:Extremities / Q000196:embryology* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D017880:Limb Deformities, Congenital; D051381:Rats; D011919:Rats, Inbred Strains; D013997:Time Factors
D004247:DNA; D001973:Bromodeoxyuridine