Antioxidant supplementation during in vitro culture improves mitochondrial function and development of embryos from aged female mice.
Maternal aging results in reduced oocyte and blastocyst quality, thought to be due, in part, to mitochondrial dysfunction and accumulation of reactive oxygen species. To reduce oxidative stress, the antioxidants α-lipoic acid (ALA; 10µM), α-tocopherol (250µM), hypotaurine (1mM) and N-acetylcysteine (NAC; 1mM), and sirtuin (100ngmL(-1)) were added to embryo culture medium (AntiOX) and compared with a control (CON) without antioxidants to assess blastocyst development after in vitro maturation and fertilisation of oocytes from aged B6D2F1 female mice (13.5 months). Development to the blastocyst stage increased in the AntiOX compared with CON group (87.6% vs 72.7%, respectively; P<0.01), in addition to higher mitochondrial membrane potential and ATP levels in the AntiOX group. Expression of genes associated with oxidative stress (PI3K, FOXO3A and GLRX2) was upregulated in the CON compared with AntiOX group. In addition to AntiOX, a medium containing only NAC and ALA (rAntiOX) was used to culture embryos from young CF1 females (6-8 weeks). More blastocysts developed in the rAntiOX compared with CON group (64.1% vs 43.3%, respectively; P<0.01), although AntiOX (48.0% blastocysts) did not result in improved development in young mice. Antioxidants improved mitochondrial activity, gene expression and development in embryos of older female mice, whereas a reduced level of antioxidants during culture was beneficial to embryos from young mice.
National Foundation for Fertility Research, 10290 RidgeGate Cr., Lone Tree, CO 80124, USA.
Elena
E
Silva
National Foundation for Fertility Research, 10290 RidgeGate Cr., Lone Tree, CO 80124, USA.
Alison F
AF
Greene
National Foundation for Fertility Research, 10290 RidgeGate Cr., Lone Tree, CO 80124, USA.
Kevin
K
Strauss
National Foundation for Fertility Research, 10290 RidgeGate Cr., Lone Tree, CO 80124, USA.
Jason R
JR
Herrick
Colorado Center for Reproductive Medicine, 10290 RidgeGate Cr., Lone Tree, CO 80124, USA.
William B
WB
Schoolcraft
National Foundation for Fertility Research, 10290 RidgeGate Cr., Lone Tree, CO 80124, USA.
Rebecca L
RL
Krisher
D000111:Acetylcysteine / Q000494:pharmacology; D000367:Age Factors; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects* / Q000502:physiology; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D051379:Mice; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D037761:Sirtuins / Q000494:pharmacology; D013654:Taurine / Q000031:analogs & derivatives / Q000494:pharmacology; D008063:Thioctic Acid / Q000494:pharmacology; D024502:alpha-Tocopherol / Q000494:pharmacology
D000975:Antioxidants; D013654:Taurine; C003949:hypotaurine; D008063:Thioctic Acid; D037761:Sirtuins; D024502:alpha-Tocopherol; D000111:Acetylcysteine