Ventricular mitochondrial gene expression during development and following embryonic ethanol exposure.
The effects of chronic embryonic ethanol exposure were evaluated in chick ventricular muscle. Ethanol treatments were administered on embryonic days 11, 13, 15, and 17 and chicks were sacrificed at various time points following treatments. Fluctuations in embryonic blood ethanol levels were examined following treatments. Developmental increases in the activities of mitochondrial enzymes, cytochrome oxidase (CO) and citrate synthase (CS), were observed. Ethanol exposure resulted in a depression in CO activity, but not CS activity. Since, a maximal depression in CO activity was seen with ethanol treatments of 75 mg/100 g, this dosing paradigm was adopted for subsequent experiments. A tissue-specific effect of ethanol was demonstrated as CO activity was unchanged in atrial, liver, pectoralis, and brain tissues. The role of mitochondrial DNA replication and transcription during the developmental up-regulation and ethanol-induced down-regulation of CO activity was evaluated using a cDNA probe for cytochrome oxidase subunit III (COIII). The relative levels of COIII mRNA and mitochondrial DNA (cpm/mg protein) decreased by 3-fold and 4-fold, respectively, across the developmental time course, while CO activity increased by 3.5-fold. Therefore, increases in mitochondrial DNA and mitochondrial mRNA transcripts are unlikely to be responsible for the developmentally-regulated increases in CO activity. Similarly, embryonic ethanol exposure failed to elicit alterations in COIII mRNA levels, indicating that the ethanol-induced depression in CO activity was not transcriptionally regulated. However, ventricular mitochondrial DNA concentrations were elevated in ethanol-treated embryos, indicating that ethanol-exposure either directly or indirectly induces mitochondrial DNA replication.
Kennedy
J M
JM
University of Illinois, Department of Physiology and Biophysics, Chicago 60640.
Kelley
S W
SW
Meehan
J M
JM
D000818:Animals; D002642:Chick Embryo; D004305:Dose-Response Relationship, Drug; D003576:Electron Transport Complex IV / Q000187:drug effects*; D005314:Embryonic and Fetal Development / Q000187:drug effects / Q000235:genetics; D000431:Ethanol / Q000097:blood; D063647:Fetal Alcohol Spectrum Disorders / Q000097:blood / Q000235:genetics*; D005786:Gene Expression Regulation / Q000502:physiology*; D006352:Heart Ventricles / Q000187:drug effects* / Q000196:embryology; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000502:physiology; D009928:Organ Specificity / Q000502:physiology; D014158:Transcription, Genetic; D016276:Ventricular Function
D000431:Ethanol; D003576:Electron Transport Complex IV