Increase of mitochondrial DNA content and transcripts in early bovine embryogenesis associated with upregulation of mtTFA and NRF1 transcription factors.
BACKGROUND
Recent work has shown that mitochondrial biogenesis and mitochondrial functions are critical determinants of embryonic development. However, the expression of the factors controlling mitochondrial biogenesis in early embryogenesis has received little attention so far.


METHODS
We used real-time quantitative PCR to quantify mitochondrial DNA (mtDNA) in bovine oocytes and in various stages of in vitro produced embryos. To investigate the molecular mechanisms responsible for the replication and the transcriptional activation of mtDNA, we quantified the mRNA corresponding to the mtDNA-encoded cytochrome oxidase 1 (COX1), and two nuclear-encoded factors, i.e. the Nuclear Respiratory Factor 1 (NRF1), and the nuclear-encoded Mitochondrial Transcription Factor A (mtTFA).


RESULTS
Unlike findings reported in mouse embryos, the mtDNA content was not constant during early bovine embryogenesis. We found a sharp, 60% decrease in mtDNA content between the 2-cell and the 4/8-cell stages. COX1 mRNA was constant until the morula stage after which it increased dramatically. mtTFA mRNA was undetectable in oocytes and remained so until the 8/16-cell stage; it began to appear only at the morula stage, suggesting de novo synthesis. In contrast, NRF1 mRNA was detectable in oocytes and the quantity remained constant until the morula stage.


CONCLUSIONS
Our results revealed a reduction of mtDNA content in early bovine embryos suggesting an active process of mitochondrial DNA degradation. In addition, de novo mtTFA expression associated with mitochondrial biogenesis activation and high levels of NRF1 mRNA from the oocyte stage onwards argue for the essential function of these factors during the first steps of bovine embryogenesis.
May-Panloup
Pascale
P
Biologie de la Reproduction, Labo FIV, Centre Hospitalier Universitaire d'Angers, 4 rue Larrey, F-49033 Angers, France. pamaypanloup@chu-angers.fr
Vignon
Xavier
X
Chrétien
Marie-Françoise
MF
Heyman
Yvan
Y
Tamassia
Manoel
M
Malthièry
Yves
Y
Reynier
Pascal
P
D000818:Animals; D002417:Cattle; D004272:DNA, Mitochondrial / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000096:biosynthesis*; D003576:Electron Transport Complex IV / Q000096:biosynthesis; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D024101:Mitochondrial Proteins / Q000096:biosynthesis*; D051266:Nuclear Respiratory Factor 1 / Q000096:biosynthesis*; D009865:Oocytes / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000096:biosynthesis*; D015854:Up-Regulation
D004272:DNA, Mitochondrial; D004268:DNA-Binding Proteins; D024101:Mitochondrial Proteins; D051266:Nuclear Respiratory Factor 1; D012333:RNA, Messenger; D014157:Transcription Factors; C422562:mitochondrial transcription factor A; D003576:Electron Transport Complex IV