Teratogen-Induced Activation of p53 in Early Postimplantation Mouse Embryos

• Published in: Toxicological sciences Vol. 95; no. 1; pp. 257 - 269
• Main Authors: Hosako, Hiromi, Little, Sally A., Barrier, Marianne, Mirkes, Philip E.
• Format: Journal Article
• Language: English
• Published: United States Oxford University Press 01.01.2007
• Subjects: 4-hydroperoxycyclophosphamide; Animals; apoptosis; Apoptosis - drug effects; Apoptosis Regulatory Proteins; Blotting, Western; Cell Cycle - drug effects; cell cycle arrest; Cyclin-Dependent Kinase Inhibitor p21 - metabolism; Cyclophosphamide - analogs & derivatives; Cyclophosphamide - toxicity; day 9 mouse embryo; Embryo Culture Techniques; Embryo, Mammalian - drug effects; Embryo, Mammalian - metabolism; Embryo, Mammalian - pathology; Embryonic Development - drug effects; embryotoxicity; Hot Temperature - adverse effects; hyperthermia; Kinetics; Mice; Noxa; p21; p53; Phosphorylation; Proto-Oncogene Proteins c-bcl-2 - metabolism; Puma; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Teratogens - toxicity; Time Factors; Tumor Suppressor Protein p53 - metabolism; Tumor Suppressor Proteins - metabolism; hyperthermia; day 9 mouse embryo; embryotoxicity; 4-hydroperoxycyclophosphamide; apoptosis; cell cycle arrest; p21; p53; Noxa; Puma
• ISSN: 1096-6080; 1096-0929
• DOI: 10.1093/toxsci/kfl143

Hyperthermia (HS) and 4-hydroperoxycyclophosphamide (4CP) activate the mitochondrial apoptotic pathway in day 9 mouse embryos. Previous microarray analyses Microarray analyses revealed that several p53 target genes are upregulated after exposure to HS or 4CP, suggesting a role for p53 in teratogen-induced apoptosis. To explore the role of p53, we assessed the activation of p53 in day 9 mouse embryos exposed to HS or 4CP in vitro. Both teratogens induced the accumulation of p53 and phosphorylation of p53 at ser-15, two hallmarks of p53 activation. HS and 4CP also induced an increase in Noxa and Puma mRNAs, transcripts of two known proapoptotic p53 target genes; however, these two teratogens did not induce significant increases in NOXA and PUMA proteins, suggesting that p53 does not activate the mitochondrial apoptotic pathway by transcriptionally upregulating the expression of NOXA and PUMA proteins. HS and 4CP also induced the expression of p21 mRNA and protein, suggesting a role for p53 in teratogen-induced cell cycle arrest. Previously, we also showed that HS and 4CP activate the apoptotic pathway in the embryo proper (head and trunk) but not in the heart. We now show that HS and 4CP induce a robust activation of p53 in the embryo proper but an attenuated induction in the heart. HS and 4CP induce the expression of p21 protein in majority of the cells in the embryo; however, expression of NOXA and PUMA proteins were not significantly induced in heads, hearts, or trunks of day 9 embryos. Overall, our results suggest that p53 may play a transcription-dependent role in teratogen-induced cell cycle arrest but a transcription-independent role in teratogen-induced apoptosis in day 9 mouse embryos exposed to HS or 4CP.