Differentially methylated CpG sites related to fertility in Japanese Black bull spermatozoa: epigenetic biomarker candidates to predict sire conception rate

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  • TAKEDA Kumiko
    Institute of Livestock and Grassland Science, NARO, Ibaraki 305-0901, Japan
  • KOBAYASHI Eiji
    Institute of Livestock and Grassland Science, NARO, Ibaraki 305-0901, Japan
  • OGATA Kazuko
    Institute of Livestock and Grassland Science, NARO, Ibaraki 305-0901, Japan
  • IMAI Akira
    Hiroshima Prefectural Livestock Technology Research Center, Hiroshima 739-0151, Japan
  • SATO Shinya
    Hiroshima Prefectural Livestock Technology Research Center, Hiroshima 739-0151, Japan
  • ADACHI Hiromichi
    Hida Beef Cattle Research, Gifu Prefectural Livestock Research Institute, Gifu 506-0101, Japan
  • HOSHINO Yoichiro
    Kyoto University, Kyoto 622-0203, Japan
  • NISHINO Kagetomo
    Beef Cattle Institute, Ibaraki Prefectural Livestock Research Center, Ibaraki 319-2224, Japan
  • INOUE Masahiro
    Tottori Prefectural Livestock Research Institute, Tottori 689-2503, Japan
  • KANEDA Masahiro
    Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan
  • WATANABE Shinya
    Institute of Livestock and Grassland Science, NARO, Ibaraki 305-0901, Japan Western Region Agricultural Research Center, NARO, Shimane 694-0013, Japan

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<p> For semen suppliers, predicting the low fertility of service bull candidates before artificial insemination would help prevent economic loss; however, predicting bull fertility through in vitro assessment of semen is yet to be established. In the present study, we focused on the methylated CpG sites of sperm nuclear DNA and examined methylation levels to screen new biomarkers for predicting bull fertility. In frozen-thawed semen samples collected from Japanese Black bulls, for which the sire conception rate (SCR) was recorded, the methylation level of each CpG site was analyzed using human methylation microarray. According to regression analysis, 143 CpG sites related to SCR were significantly differentially methylated. Whole genome bisulfite sequence data were obtained from three semen samples and the differentially methylated regions (DMRs) that included the target CpG sites selected by human methylation microarray were confirmed. Using combined bisulfite restriction analysis, fertility-related methylation changes were detected in 10 DMRs. With the exception of one DMR, the methylation levels of these DMRs were significantly different between groups with high fertility (> 50%) and low fertility (< 40%). From multiple regression analysis of methylation levels and SCR, three DMRs were selected that could effectively predict bull fertility. We suggest that these fertility-related differences in spermatozoal methylation levels could be new epigenetic biomarkers for predicting bull fertility.</p>

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