Study on the physiological protection of the reducing cryotolerance against seminal plasma prior to sperm cryopreservation in Okinawan native Agu pig

• Project/Area Number: 24580412
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied animal science
• Research Institution: University of the Ryukyus
• Principal Investigator: Tatemoto Hideki 琉球大学, 農学部, 教授 (70227114)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 精子 / 精漿 / 凍結保存 / 細胞障害 / スキムミルク / 酸化ストレス / アグー豚 / アグー精子 / L-carnitine / ミトコンドリア / 耐凍能 / Trolox / 抗酸化能 / 脂質過酸化 / カゼインナトリウム

Outline of Final Research Achievements

This study was conducted to elucidate the protective effect of skim milk (SM), casein and Trolox (TRO) added to the semen diluents on maintenance of sperm cryotolerance during semen standing incubation of Agu pig.
Treatment with 0.5% SM or casein during semen standing had the beneficial effects on post-thaw sperm motility and plasmalemma integrity. Likewise, treatment with SM plus TRO increased sperm motility, and the integrity of plasmalemma and mitochondria, and decreased the amount of lipoperoxidation, whereas these phenomena were not obviously found in sperm treated with TRO only during freezing procedure. Furthermore, higher sperm penetrability to matured oocytes in vitro was maintained in post-thaw sperm treated with SM plus TRO after the beginning of semen standing.
These findings indicate that treatment with SM or casein plus TRO during semen standing incubation improved the post-thaw qualities of Agu sperm by protection of cryotolerance prior to the freezing process.

Research Products

• [Journal Article] Addition of casein to the diluents during semen transportation improves the post-thaw qualities of Okinawan native Agu pig spermatozoa (2015)
  • Author(s): Tatemoto H, Oshiro R, Shimada H, Konno T, Yamanaka K, Ashizawa K
  • Journal Title: Journal of Warm Regional Society of Animal Science, Japan Volume: 58
  • NAID: 130005086653
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Population fluctuation and habitat preference of Ijima's Copper Pheasant <i>Syrmaticus soemmerringii ijimae</i>: an endemic, &lsquo;near threatened&rsquo; Japanese subspecies (2014)
  • Author(s): Ashizawa K, Kawaji N, Tanaka A, Nagase D, Matsumoto Y, Tatemoto H, Tsuzuki Y
  • Journal Title: ORNITHOLOGICAL SCIENCE Volume: 13 Issue: 2 Pages: 77-81
  • DOI: 10.2326/osj.13.77
  • NAID: 130004725551
  • ISSN: 1347-0558
  • Peer Reviewed
• [Journal Article] Regulation of fowl sperm motility: Evidence for the indirect, but not direct, involvement of dynein-ATPase activity on the reversible temperature-dependent immobilization. (2013)
  • Author(s): Ashizawa, K., Oyama, N., Katayama, S., Narumi, K., Tatemoto, H. and Tsuzuki, Y.
  • Journal Title: Theriogenology Volume: 79 Issue: 3 Pages: 558-565
  • DOI: 10.1016/j.theriogenology.2012.11.010
  • Peer Reviewed
• [Journal Article] Acrosome reaction of fowl sperm: evidence for shedding of acrosomal cap in intact form to release acrosomal enzyme (2013)
  • Author(s): Ahammad MU, Nishino C, Tatemoto H, Okura N, Okamoto S, Kawamoto Y, Nakada T
  • Journal Title: Poultry Science Volume: 92 Issue: 3 Pages: 798-803
  • DOI: 10.3382/ps.2012-02523
  • Peer Reviewed
• [Journal Article] Plumping fluid added to storage medium increases (2-fold) the functional life-span of fowl sperm in vitro at 4°C (2013)
  • Author(s): Ahammad MU, Nishino C, Tatemoto H, Okura N, Okamoto S, Kawamoto Y, Nakada T
  • Journal Title: British Poultry Science Volume: 54 Issue: 2 Pages: 270-280
  • DOI: 10.1080/00071668.2013.778956
  • Peer Reviewed
• [Journal Article] Persistence and accumulation of micronucleated hepatocytes in liver of rats after repeated administration of diethylnitrosamine. (2013)
  • Author(s): Narumi, K., Ashizawa, K., Fujiishi, Y., Tochinai, R., Okada, E., Tsuzuki, Y., Tatemoto, H., Hamada, S., Kaneko, K. and Ohyama, W.
  • Journal Title: Mutation Research Volume: 755 Issue: 2 Pages: 100-107
  • DOI: 10.1016/j.mrgentox.2013.03.012
  • Peer Reviewed
• [Journal Article] The involvement of N-glycosylation of zona glycoproteins during meiotic maturation in sperm-zona pellucida interactions of porcine denuded oocytes (2013)
  • Author(s): Lay KM, Nakada T, Tatemoto H
  • Journal Title: Animal Science Journal Volume: 84 Issue: 1 Pages: 8-14
  • DOI: 10.1111/j.1740-0929.2012.01027.x
  • Peer Reviewed
• [Journal Article] Effects of Fluid Secreted from the Uterus on Duration of Fertile Egg Production in Hens, and Survivability and Penetrability of Fowl Sperm <i>in vitro</i> (2013)
  • Author(s): Ahammad MU, Miyazato T, Nishino C, Tatemoto H, Okura N, Okamoto S, Kawamoto Y, Nakada T
  • Journal Title: The Journal of Poultry Science Volume: 50 Issue: 1 Pages: 74-82
  • DOI: 10.2141/jpsa.0120045
  • NAID: 130004446382
  • ISSN: 1346-7395, 1349-0486
  • Peer Reviewed
• [Journal Article] Development of a repeated-dose liver micronucleus assay using adult rats: an investigation of diethylnitrosamine and 2,4-diaminotoluene. (2012)
  • Author(s): Narumi, K., Ashizawa, K., Takashima, R., Takasawa, H., Katayama, S., Tsuzuki, Y., Tatemoto, H., Morita, T., Hayashi, M. and Hamada, S.
  • Journal Title: Mutation Res. Volume: 747 Issue: 2 Pages: 234-239
  • DOI: 10.1016/j.mrgentox.2012.05.012
  • Peer Reviewed
• [Journal Article] Improvement of the post-thaw qualities of Okinawan native Agu pig sperm frozen in an extender supplemented with antiapoptotic PTD-FNK protein (2012)
  • Author(s): Shimokawa K, Oshiro R, Yamanaka K, Ashizawa K, Ohta S, Tatemoto H
  • Journal Title: Theriogenology Volume: 78(7) Issue: 7 Pages: 1446-1455
  • DOI: 10.1016/j.theriogenology.2012.06.005
  • Peer Reviewed
• [Presentation] Improvement in the post-thaw qualities of Okinawan native Agu pig spermatozoa treated with skim milk and Trolox prior to cryopreservation (2016)
  • Author(s): Tatemoto H, Mitsube R, Tokeshi I, Uehara M, Sadoyama Y, Funabiki M, Konno T
  • Organizer: The 17th AAAP Animal Science Congress
  • Place of Presentation: Kyushu Sangyo University, Fukuoka
  • Year and Date: 2016-08-22
  • Int'l Joint Research
• [Presentation] 精液輸送ならびに精子凍結時のTrolox処理が沖縄在来豚アグー精子の融解後の性状改善に及ぼす効果 (2015)
  • Author(s): 建本秀樹，島田晴加，金野俊洋
  • Organizer: 第8回日本暖地畜産学会熊本大会
  • Place of Presentation: 東海大学熊本キャンパス
  • Year and Date: 2015-10-24
• [Presentation] 安定した美味しいアグ～豚肉の生産に向けて (2015)
  • Author(s): 建本秀樹
  • Organizer: JAおきなわ銘柄豚推進協議会定期総会
  • Place of Presentation: JAおきなわ南風原支店 会議室
  • Year and Date: 2015-09-07
  • Invited
• [Presentation] 琉球在来豚アグーにおける精液輸送用懸濁液へのカゼイン添加が凍結融解後の精子性状に及ぼす影響 (2012)
  • Author(s): 大城柳子，上地成美，建本秀樹
  • Organizer: 第105回日本繁殖生物学会大会
  • Place of Presentation: 筑波大学大学会館