Embryo mitochondrial DNA depletion is reversed during early embryogenesis in cattle.
The extensive replication of mitochondria during oogenesis and the wide variability in mitochondrial DNA (mtDNA) copy numbers present in fully grown oocytes indicate that mtDNA amount may play an important role during early embryogenesis. Using bovine oocytes derived from follicles of different sizes to study the influence of mtDNA content on development, we showed that oocytes obtained from small follicles, known to be less competent in developing into blastocysts, contain less mtDNA than those originating from larger follicles. However, because of the high variability in copy number, a more accurate approach was examined in which parthenogenetic one-cell embryos were biopsied to measure their mtDNA content and then cultured to assess development capacity. Contrasting with previous findings, mtDNA copy number in biopsies was not different between competent and incompetent embryos, indicating that mtDNA content is not related to early developmental competence. To further examine the importance of mtDNA on development, one-cell embryos were partially depleted of their mtDNA (64% +/- 4.1% less) by centrifugation followed by the removal of the mitochondrial-enriched cytoplasmic fraction. Surprisingly, depleted embryos developed normally into blastocysts, which contained mtDNA copy numbers similar to nonmanipulated controls. Development in depleted embryos was accompanied by an increase in the expression of genes (TFAM and NRF1) controlling mtDNA replication and transcription, indicating an intrinsic ability to restore the content of mtDNA at the blastocyst stage. Therefore, we concluded that competent bovine embryos are able to regulate their mtDNA content at the blastocyst stage regardless of the copy numbers accumulated during oogenesis.
Chiaratti
Marcos R
MR
Departamento de Ciências Básicas, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, Brazil. chiaramr@fcm.unicamp.br
Bressan
Fabiana F
FF
Ferreira
Christina R
CR
Caetano
Alexandre R
AR
Smith
Lawrence C
LC
Vercesi
Aníbal E
AE
Meirelles
Flávio V
FV
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002417:Cattle; D004261:DNA Replication; D004272:DNA, Mitochondrial / Q000378:metabolism*; D046149:Embryo Culture Techniques; D047108:Embryonic Development; D005260:Female; D018628:Gene Dosage; D009865:Oocytes / Q000378:metabolism*
D004272:DNA, Mitochondrial