Yang H Y, Zelus B D, Juchau M R
Department of Pharmacology, School of Medicine, University of Washington, Seattle 98195.
Biochem Biophys Res Commun. 1991 Jul 15;178(1):236-41. doi: 10.1016/0006-291x(91)91804-l.
Utilizing the reverse transcriptase-linked polymerase chain reaction, we analyzed the capacity of three groups of rat conceptal tissues to express cytochrome P4501A1 (CYP1A1) mRNA during the dysmorphogenesis-sensitive stage of organogenesis. The visceral yolk sac, ectoplacental cone and embryo proper each were investigated on day 12 of gestation with and without prior exposure in utero to 3-methylcholanthrene as inducing agent. With two sets of discriminating oligonucleotide primers, definitive, reproducible signals were detectable only in tissues from 3-methylcholanthrene preexposed conceptuses. Signals of highest intensity were observed with visceral yolk sac tissues and signals of lowest intensity were observed with tissues of the embryo per se. Specificities of the amplified cDNAs were verified using Southern blotting with hybridization to an internal oligonucleotide probe. The results indicate that organogenesis-stage conceptual tissues of the rat will express CYP1A1 mRNA in response to environmental transregulating agents.
利用逆转录酶连接的聚合酶链反应，我们分析了三组大鼠胚胎组织在器官发生的畸形发生敏感阶段表达细胞色素P4501A1（CYP1A1）mRNA的能力。在妊娠第12天，对内脏卵黄囊、外胎盘锥和胚胎本身进行了研究，有无在子宫内预先暴露于作为诱导剂的3-甲基胆蒽。使用两组鉴别性寡核苷酸引物，仅在预先暴露于3-甲基胆蒽的胚胎组织中可检测到明确的、可重复的信号。在内脏卵黄囊组织中观察到强度最高的信号，而在胚胎本身的组织中观察到强度最低的信号。使用与内部寡核苷酸探针杂交的Southern印迹法验证了扩增cDNA的特异性。结果表明，大鼠器官发生阶段的胚胎组织将响应环境反式调节剂而表达CYP1A1 mRNA。
