Hicks H E, Banes A J
Proc Soc Exp Biol Med. 1985 Dec;180(3):483-7. doi: 10.3181/00379727-180-42206.
Pregnant A/J mice received 60 mg phenytoin/kg body weight on Day 10 of gestation. Eighteen hours after phenytoin injection, animals were injected (ip) with 20 microCi/g of [35S]methionine. After 6 hr of incorporation animals were sacrificed and the embryos were removed. Protein synthesis in the embryo, as measured by [35S]methionine incorporation into trichloroacetic-precipitable protein, was analyzed by SDS-PAGE and quantitation of autoradiograms. The results of gel electrophoresis indicate that in embryonic primary palates and limb buds from phenytoin-treated mothers there is an increase in synthesis of 66-, 50-, 44-, and 13-kDa proteins and a decrease in synthesis of an 18-kDa protein compared with values for the control counterpart. No role has been assigned to the 66-, 44-, or 13-kDa proteins but the 50-kDa band comigrates with tubulin and the 18-kDa band comigrates with calmodulin. Palatal cells in vitro stained positively with specific antibody to both these proteins. An adverse effect of the anticonvulsant drug phenytoin, when administered to pregnant A/J mice is an increase in the incidence of cleft lip with or without cleft palate [CL(P)] in their offspring. These alterations in protein synthesis may be a direct or secondary result of maternal phenytoin treatment and may play a role in CL(P) formation in vivo.
怀孕的A/J小鼠在妊娠第10天接受60毫克苯妥英/千克体重的剂量。苯妥英注射18小时后，给动物腹腔注射20微居里/克的[35S]蛋氨酸。掺入6小时后处死动物并取出胚胎。通过将[35S]蛋氨酸掺入三氯乙酸沉淀蛋白来测量胚胎中的蛋白质合成，并通过SDS-PAGE和放射自显影片定量分析。凝胶电泳结果表明，与对照相比，苯妥英处理的母亲所产胚胎的原发腭和肢芽中，66、50、44和13千道尔顿蛋白质的合成增加，而18千道尔顿蛋白质的合成减少。尚未确定66、44或13千道尔顿蛋白质的作用，但50千道尔顿条带与微管蛋白共迁移，18千道尔顿条带与钙调蛋白共迁移。体外腭细胞用针对这两种蛋白质的特异性抗体染色呈阳性。抗惊厥药物苯妥英对怀孕的A/J小鼠给药时，其后代唇裂伴或不伴腭裂[CL(P)]的发生率会增加。这些蛋白质合成的改变可能是母体苯妥英治疗的直接或间接结果，并且可能在体内CL(P)的形成中起作用。
