Mitochondrial function and intracellular distribution is severely affected in in vitro cultured mouse embryos

Abstract

AbstractStudies of mitochondrial dynamics have identified an intriguing link between energy supply balance and mitochondrial architecture. This suggests that inappropriate culture conditions might inhibit mitochondrial functions, and affect embryonic development. Therefore, this study was conducted to determine whether in vitro culture (IVC) might affect mitochondrial function, distribution, organization (by Mitotracker Green), gene expression on RNA level (by qPCR), and protein expression and localization (by western blot and immunostaining) involved in regulation of mitochondrial functions. Mitochondria in 2-cell IVC embryos were less numerous compare to IN VIVO while the localization and distribution do not differ between the groups. Mitochondria of in vivo blastocysts formed elongated network along the cells, while in IVC were fragmented, rounded, and aggregated mainly in the perinuclear region. Additionally, mitochondria of IN VIVO embryos moved back and forth along their long axis on radial tracks, while in IVC blastocysts were much less active. mtDNA copy number in IVC blastocysts (92,336.65 ± 5860.04) was significantly lower than that of IN VIVO (169,103.92 ± 16,322.41; P < 0.02) as well as lower protein expressions responsible for mitochondrial fusion was observed in IVC blastocysts. Results indicate that in vitro culture affect on perturbations in mitochondrial number and function, which is associated with decreased developmental competence of in vitro produced mouse embryos.