13.1% of the world’s population were undernourished in 2010. Poverty and the resultant undernourishment are root causes of parental malnutrition in developing countries, which ultimately contributes to the generation of unhealthy offspring. However, whether supplementing undernourished males with vitamins and antioxidants can reverse aberrant sperm parameters in the next generation remains unknown.
C57BL6 male mice (n=30, 6 weeks old) were fed either ad libitum control diet (CD), diet restricted CD (DR), or DR supplemented with vitamins and antioxidants (DR+AOx). DR/DR+AOx cohorts were restricted to 90%, 80%, and 70% of the food intake of CD mice by weight for the first 3 weeks, then maintained at 70% for a further 11 weeks. Males were subsequently mated with ad libitum CD females to produce offspring. Offspring sperm samples (n=5 per group, with 3-4 fathers represented, 9 weeks old) were used for sperm binding and stained for total reactive oxygen species (ROS; DCFDA) and mitochondrial ROS (MitoSOX Red).
Offspring fathered by DR males had significantly reduced number of sperm bound to an oocyte (30.48±1.60, p<0.01) compared with offspring of CD fathers (60.21±2.16), while DR+AOx offspring had sperm binding restored to control level (58.34±1.82). Significant decreases in sperm both total and mitochondrial ROS were observed in offspring sired by DR males (378.18±17.25, p<0.01 and 32.13%±7.12, p<0.05) and further reduced in offspring of DR+AOx fathers (237.89±7.00, p<0.01 and 24.84%±5.10, p<0.02) compared to CD offspring (438.76±13.00 and 72.75%±10.59).
Thus impaired sperm binding in offspring arising from paternal undernutrition can be ameliorated by paternal vitamin and antioxidant supplementation. Interestingly, paternal diet restriction alone reduced sperm ROS levels in offspring which were further decreased by vitamin and antioxidant supplementation during diet restriction. This data provides the first evidence that dietary vitamin and antioxidant supplementation to malnourished fathers prior to conception could significantly improve reproductive outcomes in male offspring.