Characterization of mitochondrial replication and transcription control during rat early development in vivo and in vitro

Abstract

In vitroculture (IVC), used in assisted reproductive technologies, is a major environmental stress on the embryo. To evaluate the effect of IVC on mitochondrial transcription and the control of mtDNA replication, we measured the mtDNA copy number and relative amount of mRNA for mitochondrial-related genes in individual rat oocytes, zygotes and embryos using real-time PCR. The average mtDNA copy number was 147 600 (±3000) in metaphase II oocytes. The mtDNA copy number was stable throughoutin vivoearly development and IVC induced an increase in mtDNA copy number from the 8-cell stage onwards.GapdmRNA levels vary during early development and IVC did not change the patterns of these housekeeping gene transcripts.PolrmtmRNA levels did not vary during early development up to the morula stage but increased at the blastocyst stage. IVC induced the up-regulation ofPolrmtmRNA, one of the key genes regulating mtDNA transcription and replication, at the blastocyst stage. An increase inmt-Nd4mRNA preceded the blastocyst-related event observed in nuclear-encodedGapdandPolrmt, suggesting that the expression of mitochondrial encoded genes is controlled differently from nuclear encoded genes. We conclude that the IVC system can perturb mitochondrial transcription and the control of mtDNA replication in rat embryos. This perturbation of mtDNA regulation may be responsible for the abnormal physiology, metabolism and viability ofin vitro-derived embryos.