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Ethanol Elevates c‐Myc Levels in Cultured Mouse Preimplantation Embryos
Author(s) -
Leach Richard E.,
Rout Ujjwal K.,
Schultz Jeffrey F.,
Saunders Dwight E.,
Armant D. Randall
Publication year - 1999
Publication title -
alcoholism: clinical and experimental research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.267
H-Index - 153
eISSN - 1530-0277
pISSN - 0145-6008
DOI - 10.1111/j.1530-0277.1999.tb04183.x
Subject(s) - blastocyst , embryo , microbiology and biotechnology , messenger rna , western blot , biology , andrology , embryogenesis , gene expression , ethanol , chemistry , biochemistry , gene , medicine
A brief exposure to ethanol accelerates the rate of early mouse embryonic development in vitro, increasing blastocyst formation, trophohlast outgrowth, and implantation rates after embryo transfer. The physiological effects of ethanol during preimplantation development are associated with rapid changes in gene expression and apparently arise from the ability of ethanol to elevate cytoplasmic free Ca 2+ and alter cellular signaling pathways. The purpose of this study was to examine whether the abundance of c‐Myc, a transcription factor that promotes cell proliferation and is required for blastocyst development, is upregulated in mouse blastocysts challenged with ethanol. After exposure of mouse blastocysts to 0.1% (17.5 mM) ethanol, we determined the levels of: 1) c‐Myc mRNA, using reverse transcription and the polymerase chain reaction; and 2) c‐Myc protein levels, using specific monoclonal antibodies. Within 10 min of exposure to ethanol, the relative abundance of c‐Myc mRNA increased 6‐fold, then rapidly returned to baseline levels within 1 hr. As expected, elevation of c‐Myc mRNA by ethanol was attenuated in embryos that were first treated with the intracellular Ca 2+ chelator, BAPTA‐AM. Western blot analysis of solubilized embryos revealed that c‐Myc mRNA was translated into a single 62‐kD protein that increased in intensity 30 min after treatment with ethanol. Immunocytochemical staining demonstrated that c‐Myc was localized exclusively in nuclei and that staining intensity increased significantly after 10 min. Peak levels of c‐Myc protein were found 30 min after ethanol exposure and persisted for at least 2 hr. The c‐myc proto‐oncogenc seems to be an immediate early response gene for ethanol that may regulate the transcription of other genes that influence early embryogenesis and growth.