The relative semi-quantification of mRNA expression as a useful toxicological endpoint for the identification of embryotoxic/teratogenic substances

被引:29
作者
Bigot, K
De Lange, J
Archer, G
Clothier, R
Bremer, S
机构
[1] Commiss European Communities, Joint Res Ctr, ECVAM, Inst Hlth & Consumer Protect, I-21020 Ispra, Italy
[2] Univ Nottingham, Sch Biomed Sci, FRAME Alternat Lab, Nottingham NG7 2RD, England
关键词
embryonic stem cells; teratogenicity; embryotoxicity; PCR analysis;
D O I
10.1016/S0887-2333(99)00027-2
中图分类号
R99 [毒物学(毒理学)];
学科分类号
100405 ;
摘要
Embryonic stem cells, which resemble the undifferentiated cells of the epiblast in a blastocyst, are able to differentiate into derivatives of the primary germ layers, including cardiomyocytes. The effects of embryotoxic/teratogenic compounds on the differentiation of cells was examined by semiquantification of the mRNA expression using the RT-PCR protocol. Alpha and beta myosin heavy chain (alpha-MHC and beta-MHC, respectively) mRNA expression were chosen as tissue-specific markers, characteristic of early cardiac muscle development. Nine chemicals were investigated, chosen according to their in vivo embryotoxic/teratogenic potential in mice. The teratogens all-trans retinoic acid (RA), 5-fluorouracil (5-FU), hydroxyurea (HU), diphenylhydantoin (DPH) and caffeine (Caff) caused a significant reduction in MHC mRNA expression at a dose lower than that required for cytotoxicity. Saccharin (Sacc) had a similar effect, while penicillin G (PenG), isoniazid (Iso) and cytarabine (Ara-C) only showed an effect on MHC mRNA expression when the cells had a significant loss in viability. The inability to identify the strong teratogen Ara-C is related to the inappropriate target tissue. (C) 1999 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:619 / 623
页数:5
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