Effect of Arachidonic Acid Supplement in both Maternal and Paternal Line on Global DNA Methylation in Mice transgenerationally
Authors: De la Rocha Carmen , Rodriguez‐Rios Dalia , Alvarado‐Caudillo Yolanda , Galvan Pedro , Orozco‐Castellanos Luis Manuel , Zaina Silvio , Lund Gertrud 
Published: November 2023

Ever since the developmental origins of health and disease hypothesis was proposed, there has been a growing interest in studying the effect of nutritional factors in early stages of development. It is well established that both maternal and paternal nutrition have a strong effect on the health of the offspring and this phenotype can be exacerbated in a transgenerational way epigenetically regulated. Specific polyunsaturated fatty acids (PUFAs)‐rich supplements in early stages of development have been described as beneficial for metabolic health. Thus, the aim of this work was to evaluate the effect of Arachidonic Acid (AA) supplementation for three generations on the global DNA methylation of the offspring in both paternal and maternal line.

Methods

12 week old BALB/C mice were supplemented with 37 mg/kg/day (0.3% of Fatty Acids FA) of Arachidonic Acid (♀AA F0 n=29) dissolved in soybean oil (SBO) or just SBO (♀SBO F0, n=30) as a single bolus during the pregnancy for the maternal line experiment, or during 10 days prior of matting in males for the paternal line experiment (♂AA F0 n=30, and ♂SBO F0, n=30), also a control group was analyzed with no supplement (US n=30), at weaning offspring (F1) where weighted and sexed, and 5 males and 5 females of each group where sacrificed by cervical dislocation and organs were removed and weighted (intestine, brain, liver, testis, ovaries and uterus). When progeny (F1) reached 12 weeks old, female offspring of mother supplemented received the same treatment as their mothers (♀AA F1 n=7; ♀SBO F1, n=8), and male progeny of father supplemented receive same treatment as their fathers (♂AA F1 n=7; and ♂SBO F1, n=9) and so on for another generation (♀AA F2 n=9; ♀SBO F2, n=5; ♂AA F2 n=7; and ♂SBO F2, n=9). Global DNA methylation status in liver and brain was performed using the Methylflash Methylated DNA Quantification Kit. ANOVA test were used for means comparison, and Pearson's for correlation analysis, p<0.05 was considered significant.

Results

A significant increase in the weight of ♀AA F2 and ♂AA F3, regardless of litter size (r=0.07, p=0.98), significant differences were observed in the wet weight of brain of treatment groups (p=4.60E‐10) that showed a sex‐specific susceptibility since only females showed difference, intestine (p=3.64 E‐06) and liver (p=1.25E‐11) also showed differences in the treatment groups compared to the US, being heavier in the progeny of the female line. Variations in the levels of global DNA methylation in the liver were observed among groups that correlated with organ weight (liver) also in a sex‐specific manner (r=0.6, p=0.02).

Conclusions

AA supplementation in the periconceptional period has an effect on the weight of the offspring and their organs exacerbated transgenerationally that correlates with the levels of global methylation in liver of sex‐specific manner and whose effect is greater in the maternal line.

Support or Funding Information

Fellowship from the Mexican Council for Science and Technology (CONACyT, Fellowship no. 347418)