Mitochondrial activity and morphology in developing porcine oocytes and pre-implantation non-cultured and cultured embryos.
Mitochondria are important determinants of developmental competence for oocytes and embryos owing to their central role in cellular metabolism, yet mitochondrial activity and morphometry during early porcine development have not been quantified. In this study, we examined the membrane potential Δψ(m) and the surface density Sv(in,m) of the inner mitochondrial membrane in pig oocytes and pre-implantation embryos using fluorescent probes and confocal microscopy. Mitochondria and their cristae were also examined by transmission electron microscope. Δψ(m) was consistently low from immature oocytes up to morulae and increased significantly in the early blastocyst before decreasing at the expanded blastocyst stage. This stage-dependent pattern of Δψ(m) changes differs from that reported for other mammals. We also determined that Δψ(m) is lower in cultured when compared to non-cultured porcine early blastocysts. Sv(in,m) was higher in immature oocytes than mature oocytes and remained constant up to the 4- to 8-cell embryo stage. It increased significantly at morula and early blastocyst stages. No differences in Sv(in,m) were found between developmentally matched non-cultured and cultured embryos. These results indicate that the inner mitochondrial membrane potential and surface density change significantly during pre-implantation porcine development in relation to metabolic alterations of the embryo. It is possible that modification of Δψ(m) by manipulating culture conditions may improve the performance of embryos that develop in vitro.
Romek
M
M
Department of Cytology and Histology, Institute of Zoology, Jagiellonian University, Kraków, Poland. marek.romek@uj.edu.pl
Gajda
B
B
Rolka
M
M
Smorąg
Z
Z
D000818:Animals; D001755:Blastocyst / Q000648:ultrastructure*; D046149:Embryo Culture Techniques / Q000662:veterinary*; D005260:Female; D053078:Membrane Potential, Mitochondrial; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000502:physiology* / Q000648:ultrastructure*; D051336:Mitochondrial Membranes / Q000502:physiology / Q000648:ultrastructure; D009865:Oocytes / Q000648:ultrastructure*; D034421:Sus scrofa; D013552:Swine