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Studies on the Effect of Magnesium and Polymyxin B on Liquid-stored Boar Spermatozoa

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Title Studies on the Effect of Magnesium and Polymyxin B on Liquid-stored Boar Spermatozoa( 内容と審査の要旨(Summary) )

Author(s) Quzi Sharmin Akter

Report No.(Doctoral Degree) 博士(獣医学) 甲第537号 Issue Date 2019-03-13 Type 博士論文 Version ETD URL http://hdl.handle.net/20.500.12099/77968 ※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。

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                          Ặྡ㸦ᮏ㸦ᅜ㸧⡠㸧   4X]L6KDUPLQ$NWHU㸦ࣂࣥࢢࣛࢹࢩࣗேẸඹ࿴ᅜ㸧 ୺ ᣦ ᑟ ᩍ ဨ Ặ ྡ    ᒱ㜧኱Ꮫ ᩍᤵ ᮧ ℩ ဴ ☻ Ꮫ ఩ ࡢ ✀ 㢮    ༤ኈ㸦⋇་Ꮫ㸧 Ꮫ ఩ グ ␒ ྕ    ⋇་༤⏥➨㸳㸱㸵ྕ Ꮫ ఩ ᤵ ୚ ᖺ ᭶ ᪥   ᖹᡂ㸱㸯ᖺ㸱᭶㸯㸱᪥     Ꮫ ఩ ᤵ ୚ ࡢ せ ௳    Ꮫ఩つ๎➨㸲᮲➨㸯㡯ヱᙜ  ◊ ✲ ⛉ ཬ ࡧ ᑓ ᨷ    㐃ྜ⋇་Ꮫ◊✲⛉             ⋇་Ꮫᑓᨷ ◊✲ᣦᑟࢆཷࡅࡓ኱Ꮫ   ᒱ㜧኱Ꮫ Ꮫ ఩ ㄽ ᩥ 㢟 ┠  6WXGLHVRQWKH(IIHFWRI0DJQHVLXPDQG3RO\P\[LQ %RQ/LTXLGVWRUHG%RDU6SHUPDWR]RD 㸦ࣈࢱᾮ≧ಖᏑ⢭Ꮚ࡬ཬࡰࡍ࣐ࢢࢿࢩ࣒࢘ཬࡧ࣏࣑ࣜ࢟ ࢩࣥ%ࡢຠᯝ࡟㛵ࡍࡿ◊✲㸧 ᑂ ᰝ ጤ ဨ    ୺ᰝ  ᒱ 㜧 ኱ Ꮫ  ෸ᩍᤵ  㰻 ⸨ ṇ୍㑻             ๪ᰝ  ᖏᗈ␆⏘኱Ꮫ  ᩍ ᤵ  ᯇ ஭ ᇶ ⣧             ๪ᰝ  ᒾ ᡭ ኱ Ꮫ  ᩍ ᤵ  㧘 ᶫ   ㏱             ๪ᰝ  ᮾி㎰ᕤ኱Ꮫ  ᩍ ᤵ  Ώ ㎶   ඖ             ๪ᰝ  ᒱ 㜧 ኱ Ꮫ  ᩍ ᤵ  ᮧ ℩ ဴ ☻   Ꮫ఩ㄽᩥࡢෆᐜࡢせ᪨  ㇜ࡢ⦾Ṫ࡟࠾࠸࡚ேᕤᤵ⢭ᢏ⾡ࡀᬑཬࡋ࡚ࡁ࡚࠸ࡿࡀ㸪ኟᏘ୙ዷ⑕ࡀྂࡃ࠿ࡽୡ⏺ⓗ࡟ ▱ࡽࢀ࡚࠸ࡿࠋኟᏘ࡟᥇ྲྀࡉࢀࡿ㇜⢭Ꮚࡣ㸪࢝ࣝࢩ࣒࢘ཬࡧ࢝ࣝࢩ࣒࢘࢖࢜ࣀ࣍࢔ A23187 㸦Ca2+/A23187㸧࡟཯ᛂࡋ࡚ㄏ㉳ࡉࢀࡿඛయ཯ᛂࡀ㐣ᩄ࡟Ⓨ⏕ࡋ㸪ࡇࡢࡇ࡜࡜ኟᏘ࡟⢭₢୰ ࡢ࣐ࢢࢿࢩ࣒࢘࢖࢜ࣥ㸦Mg2+㸧⃰ᗘࡀపࡃ࡞ࡿഴྥࡀ࠶ࡿࡇ࡜ཬࡧேᕤᤵ⢭ᚋࡢศፔ⋡ࡀ పୗࡍࡿࡇ࡜ࡀ㛵㐃ࡋ࡚࠸ࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋࡲࡓ㸪ኟᏘ࡟ࡣ⣽⳦ࡢቑṪࡀ᪩ࡃ࡞ ࡿࡇ࡜ࡀ⪃࠼ࡽࢀࡿࠋኟᏘࡢཷ⫾ᡂ⦼ࡀపୗࡍࡿࡇ࡜ࢆ㜼Ṇࡍࡿࡇ࡜ࢆ┠ⓗ࡜ࡋ㸪ᮏ◊✲ ࡣ෭ⶶಖᏑ⢭ᾮ࡬ῧຍ๣㸪Mg2+ ཬࡧᢠ⏕≀㉁࡛࠶ࡿ◲㓟࣏࣑ࣜ࢟ࢩࣥ B㸦PMB㸧㸪ࢆῧຍ ࡍࡿࡇ࡜࡟ࡼࡾ㐣ᩄ࡞ඛయ཯ᛂㄏ㉳ࢆᢚไࡍࡿ࠿ྰ࠿ཬࡧ PMB ࡢ⢭Ꮚࡢඛయ཯ᛂ࡬ཬ ࡰࡍᙳ㡪ࡀ࠶ࡿ࠿ྰ࠿࡟ࡘ࠸࡚ゎᯒࡋࡓࠋඛయ཯ᛂࡣཷ⢭࡟࠾ࡅࡿᚲせ୙ྍḞ࡞ࣉࣟࢭࢫ ࡛࠶ࡿࡢ࡛㸪Mg2+ ཬࡧ PMB ࡀ┤᥋⢭Ꮚᶵ⬟࡬ཬࡰࡍᙳ㡪ࢆㄪ࡭ࡓᚋ㸪୧⪅ࡢ෭ⶶಖᏑ ⢭ᾮ࡬ࡢῧຍࡀಖᏑᚋࡢ⢭Ꮚࡢ୍⯡ᛶ≧ཬࡧඛయ཯ᛂ࡬ཬࡰࡍᙳ㡪ࢆゎᯒࡋࡓࠋ  ࢝ࣝࢩ࣒࢘ࡢὶධࡀඛయ཯ᛂࡢ㛤ጞ࡟ᚲ㡲࡛࠶ࡿࡀ㸪Mg2+ ࡣ࢝ࣝࢩ࣒࢘࢖࢜ࣥ㸦Ca2+ ࡢ⏕⌮ⓗ࡞ࡢᣕᢠ๣࡛࠶ࡿࠋ➨ 2 ❶➨ 1 ⠇ࡢ◊✲ࡣ㸪Ca2+/A23187 ࡟ࡼࡾㄏ㉳ࡉࢀࡿඛ య཯ᛂ࡬ཬࡰࡍ Mg2+ ࡢᙳ㡪ࢆㄪ࡭ࡓࠋ෭ⶶಖᏑࡋࡓࣈࢱ⢭ᏊࢆὙίࡋ㸪✀ࠎࡢ␗࡞ࡿ

MgCl2 ࠶ࡿ࠸ࡣ MgSO4 ࡢᏑᅾୗ࡛ Ca2+/A23187 ࡟่࡚⃭ࡋࡓࠋ0–3 mM ࡢ MgSO4 ࠶ࡿ

࠸ࡣ MgCl2 ࡢᏑᅾୗ࡛㸪⢭Ꮚࢆ 10 ཬࡧ 15 ศ㛫่⃭ᚋࡢඛయ཯ᛂㄏ㉳⋡ࡣ㸪࠸ࡎࢀࡢ ᫬㛫࡟࠾࠸࡚ࡶ௚ࡢ⃰ᗘ࡜ẚ࡭࡚ 0.1 mM ࡢ MgCl2 ࠶ࡿ࠸ࡣ MgSO4 ࡛᭷ព࡟పୗࡋࡓࠋ ⢭Ꮚࢆ 0–0.1 mM MgSO4 ࡢᏑᅾୗ࡛ Ca2+/A23187 ࡟ࡼࡾ 60 ศ㛫ࡲ่࡛⃭ࡋࡓ⤖ᯝ㸪 Ca2+/A23187 ࡟ࡼࡾㄏ㉳ࡉࢀࡓඛయ཯ᛂࡣ MgSO 4 ࡟ࡼࡾ⃰ᗘ౫Ꮡⓗ࡟ᢚไࡉࢀ㸪ࡑࡢᢚ ไࡣ 0.1 mM ࡛᭱኱࡛࠶ࡗࡓࠋྠᵝ࡟ 0.1–3 mM ࡢ MgSO4 Ꮡᅾୗ࡛⢭Ꮚࢆ่⃭ࡋࡓ⤖ᯝ㸪 㸦㸰㸮㸧

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ࡑࡢᢚไࡣ 0.1 mM ࡛᭱኱࡛࠶ࡗࡓࠋ௨ୖࡼࡾ μM ࣞ࣋ࣝࡢ Mg2+ ࡟ࡼࡾ mM ࣞ࣋ࣝࡢ Ca2+ Ꮡᅾୗ࡛ Ca2+/A23187 ࡟ࡼࡾㄏ㉳ࡉࢀࡿඛయ཯ᛂࡣᢚไࡉࢀࡿࡇ࡜ࡀ♧၀ࡉࢀࡓࠋ  ➨ 2 ❶➨ 2 ⠇ࡢ◊✲࡛ࡣ㸪ᕼ㔘ᾮ࡛ᕼ㔘ࡋࡓࣈࢱ⢭ᾮ࡟ Mg2+ ࢆῧຍࡋ࡚ಖᏑࡋࡓሙ ྜࡢಖᏑᚋࡢ⢭Ꮚࡢ㐠ືᛶཬࡧඛయ཯ᛂ࡬ཬࡰࡍᙳ㡪ࢆㄪ࡭ࡓࠋMgSO4 (0–8 mM) ࡢῧຍ ࡛ࡣಖᏑᚋࡢ⢭Ꮚࡢ㐠ືᛶ࡟ᙳ㡪ࡋ࡞࠿ࡗࡓࡀ㸪ಖᏑࡋࡓ⢭Ꮚࡢඛయ཯ᛂࢆὙίᚋ Ca2+/A23187 ࡟ࡼࡾ่⃭ࡋࡓ⤖ᯝ㸪่⃭ࡢ 10 ཬࡧ 15 ศ࡟࠾࠸࡚ࡑࢀࡒࢀ 4 mM ῧຍࡢ ⢭Ꮚ࡟࠾࠸࡚௚ࡢ⃰ᗘ࡟ẚ࡭᭷ព࡟ඛయ཯ᛂㄏ㉳⋡ࡀపୗࡋࡓࠋ่⃭⤊஢ᚋࡢ⢭Ꮚࡢ㐠ື ⋡࡜⏕Ꮡ⋡ࡣ MgSO4 ῧຍ࡟ࡼࡿኚ໬ࡀぢࡽࢀ࡞࠿ࡗࡓࡇ࡜࠿ࡽ㸪ㄏ㉳ࡉࢀࡓඛయ཯ᛂࡣ ኚᛶ࡟ࡼࡿඛయࡢ႙ኻ࡛ࡣ࡞࠸ࡇ࡜ࡀ☜ㄆࡉࢀࡓࠋ௨ୖࡢ⤖ᯝࡣ㸪ኟᏘ୙ዷ⑕࡟≉ᚩⓗ࡛ ࠶ࡿ⢭Ꮚࡢ㐣๫࡞ඛయ཯ᛂㄏ㉳ࢆ⦆࿴࡛ࡁࡿ᪉ἲࡢ㛤Ⓨ࡟᭷⏝࡞᝟ሗࢆᥦ౪ࡋ࡚࠸ࡿࠋ  PMB ࡣ࢚ࣥࢻࢺ࢟ࢩࣥࢆ୰࿴ࡍࡿࡇ࡜࡟ࡼࡾࣈࢱ⢭ᾮࡢಖᏑ࡟᭷ຠ࡛࠶ࡿࡇ࡜ࡀሗ࿌ ࡉࢀ࡚࠸ࡿࠋࡋ࠿ࡋ㸪య⣽⬊࡟࠾࠸࡚ࡣࢩࢢࢼࣝኚ᥮ᶵᵓ࡟㛵୚ࡍࡿ཯ᛂ⤒㊰ࢆㄪ⠇ࡍࡿ ࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋ➨ 3 ❶➨ 1 ⠇ࡢᐇ㦂࡛ࡣ㸪PMB ࡀ┤᥋⢭Ꮚ࡬ཬࡰࡍᙳ㡪ࢆㄪ࡭ ࡓࠋ෭ⶶಖᏑࡋࡓ⢭ᏊࢆὙίࡋ㸪0–100 μM ࡢ PMB ࡢᏑᅾୗ࡛⢭Ꮚࢆᇵ㣴ࡋࡓ⤖ᯝ㸪㐠ື ⋡࡜⏕Ꮡ⋡ࡣ 0–50 μM ࡢ⠊ᅖ࡛ኚ໬ࡋ࡞࠿ࡗࡓࡀ㸪75 μM ཬࡧ 100 μM ࡢ PMB ῧຍ࡛ ᭷ព࡟పୗࡋࡓࠋ┤㐍㐠ື⋡ࡣ 25 μM ௨ୖࡢ㸪άⓎࡉࡣ 75 μM ௨ୖࡢ PMB ῧຍ࡟ࡼࡾ ࡑࢀࡒࢀ᭷ព࡟పୗࡋࡓࡀ㸪จ㞟ࡣ࠸ࡎࢀࡢ⃰ᗘ࡟࠾࠸࡚ࡶᙳ㡪ࢆཷࡅ࡞࠿ࡗࡓࠋ୍᪉㸪 ྠᵝ࡟Ὑίࡋࡓ⢭Ꮚࢆ㐠ື⋡࡜⏕Ꮡ⋡࡟ᙳ㡪ࡋ࡞࠿ࡗࡓ⃰ᗘ࡛࠶ࡿ 0–50 μM ࡢ PMB Ꮡ ᅾୗ࡛๓ᇵ㣴ࡋࡓᚋ㸪Ca2+/A23187 ࡟ࡼࡾඛయ཯ᛂࢆ่⃭ࡋࡓ⤖ᯝ㸪0.01–0.5 μM ཬࡧ 1– 50 μM ࡢῧຍ࡛ࡑࢀࡒࢀ่⃭ࡢ 10 ศཬࡧ 15 ศ࡛ㄏ㉳ࡉࢀࡓඛయ཯ᛂࡀቑᙉࡉࢀࡓࠋ  ➨ 3 ❶➨ 2 ⠇ࡢ◊✲࡛ࡣ㸪✀ࠎࡢ⃰ᗘ㸦0–75 μM㸧ࡢ PMB ࢆῧຍࡋ࡚ᕼ㔘ᚋಖᏑࡋ ࡓ⢭Ꮚࡢ㐠ືᛶࢆㄪ࡭ࡿ࡜ඹ࡟ PMB ῧຍࡀඛయ཯ᛂ࡬ཬࡰࡍᙳ㡪ࢆㄪ࡭ࡓࠋಖᏑࡋࡓ⢭ Ꮚࡢ㐠ື⋡ࡣ PMB ࡢῧຍ࡟ࡼࡗ࡚ࡣ᭷ព࡞ᙳ㡪ࢆཷࡅ࡞࠿ࡗࡓࡀ㸪┤㐍㐠ື⋡ࡀ PMB ࡢῧຍ⃰ᗘࡢୖ᪼࡜ඹ࡟ୖ᪼ࡋࡓࠋಖᏑࡋࡓ⢭ᏊࢆὙίᚋ㸪ྠᵝ࡟ 30 ศࡲ࡛ Ca2+/A23187 Ꮡᅾୗ࡛ᇵ㣴ࡋඛయ཯ᛂࢆ่⃭ࡋࡓࠋᇵ㣴⤊஢ᚋ㸪☜ㄆࡢ┠ⓗ࡛㸪⢭Ꮚࡢ㐠ື⋡㸪┤㐍㐠 ື⋡㸪⏕Ꮡ⋡㸪άⓎࡉཬࡧจ㞟ࢆㄪ࡭ࡓࠋᇵ㣴 15 ศཬࡧ 30 ศ࡟࠾࠸࡚ࡑࢀࡒࢀ 50 ཬ ࡧ 75 μM ࡢ PMB ῧຍ࡟ࡼࡾ Ca2+/A23187 ࡟ࡼࡾㄏ㉳ࡉࢀࡓඛయ཯ᛂࡀቑᙉࡉࢀࡓࠋ25 μM ࡢ PMB ࡢῧຍ࡟ࡼࡾ㐠ື⋡ཬࡧ⏕Ꮡ⋡ࡀ௚ࡢ⃰ᗘ࡟ẚ࡭࡚᭷ព࡟పୗࡋࡓࡀ㸪ࡇࢀ ࡽࡢ௨እࡢ⢭Ꮚࡢᛶ≧ࡣ PMB ῧຍ࡟ࡼࡾᙳ㡪ࢆཷࡅ࡞࠿ࡗࡓࠋ௨ୖࡼࡾ㸪ࣈࢱ⢭ᾮࢆ PMB Ꮡᅾୗ࡛ಖᏑࡍࡿࡇ࡜ࡀ࡛ࡁ㸪ࡇࢀ࡟ࡼࡾ㐠ື⋡࡜⏕Ꮡ⋡ࡣ୙ኚ࡛┤㐍㐠ື⋡ࡀୖ ᪼ࡋඛయ཯ᛂࡢㄏ㉳ࡀቑᙉࡉࢀࡿࡇ࡜ࡀ♧ࡉࢀࡓࠋ  ௨ୖࡼࡾ㸪࣐ࢢࢿࢩ࣒࢘ࡣ㐠ືᛶࢆᦆ࡞࠺ࡇ࡜࡞ࡃࣈࢱ෭ⶶಖᏑ⢭ᾮ࡬ῧຍࡍࡿࡇ࡜ࡀ ࡛ࡁ㸪ࡲࡓ㸪ῧຍ࡟ࡼࡾඛయ཯ᛂࡢㄏ㉳ࡀᢚไࡉࢀࡿࡇ࡜࠿ࡽ㐣๫࡞ඛయ཯ᛂࡢㄏ㉳ࢆ⦆ ࿴ࡋ㸪ኟᏘ࡟᥇ྲྀࡉࢀࡿ⢭Ꮚࡢཷ⢭⬟ຊࢆ㧗ࡵࡽࢀࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ୍᪉㸪PMB ࢆ ಖᏑ⢭ᾮ࡬ῧຍࡍࡿࡇ࡜࡟ࡼࡾཷ⢭㒊఩ࡲ࡛ࡢ⢭Ꮚࡢ⛣⾜࡟ᚲせ୙ྍḞ࡞㐠ື࡛࠶ࡿ┤㐍 㐠ື⋡ࢆ㧗ࡵࡿࡇ࡜ࡀ࡛ࡁ㸪ྠ᫬࡟༸Ꮚ࡬ࡢ౵ධ࡟ᚲせ୙ྍḞ࡞ඛయ཯ᛂㄏ㉳⬟ຊࡀ㧗ࡵ ࡽࢀ㸪ཷ⢭⬟ຊࢆୖ᪼ࡉࡏᚓࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋࡋ࠿ࡋ㸪ኟᏘ࡟᥇ྲྀࡉࢀࡿ⢭Ꮚ࡬ῧ ຍࡍࡿࡇ࡜࡟ࡼࡾ㸪㐣๫࡟ㄏ㉳ࡉࢀࡿඛయ཯ᛂࢆࡉࡽ࡟ቑᙉࡍࡿྍ⬟ᛶࡀ࠶ࡾ㸪ኟᏘ࡟౑ ⏝ࡍࡿ㝿࡟ࡣ⪃៖ࡀᚲせ࡜ᛮࢃࢀࡓࠋ ᑂ ᰝ ⤖ ᯝ ࡢ せ ᪨  ㇜ࡢ⦾Ṫ࡟࠾࠸࡚ேᕤᤵ⢭ᢏ⾡ࡀᬑཬࡋ࡚ࡁ࡚࠸ࡿࡀ㸪ᮏ◊✲ࡣྂࡃ࠿ࡽୡ⏺ⓗ࡟▱ࡽ ࢀ࡚࠸ࡿኟᏘ୙ዷ⑕ࡢᨵၿࢆᮏ◊✲ࡢ᭱⤊┠ᶆ࡜ࡋ࡚⾜ࢃࢀࡓࠋኟᏘࡢཷ⫾ᡂ⦼ࡀపୗࡍ ࡿࡇ࡜ࢆ㜼Ṇࡍࡿࡇ࡜ࢆ┠ⓗ࡜ࡋ㸪ᮏ◊✲ࡣ෭ⶶಖᏑ⢭ᾮ࡬ῧຍ๣㸪Mg2+ ཬࡧ࢚ࣥࢻࢺ

(4)

࢟ࢩࣥࢆ୰࿴ࡍࡿᢠ⏕≀㉁࡛࠶ࡿ◲㓟࣏࣑ࣜ࢟ࢩࣥ B㸦PMB㸧㸪ࢆῧຍࡍࡿࡇ࡜࡟ࡼࡾ㐣 ᩄ࡞ඛయ཯ᛂㄏ㉳ࢆᢚไࡍࡿ࠿ྰ࠿ཬࡧ PMB ࡢ⢭Ꮚࡢඛయ཯ᛂ࡬ཬࡰࡍᙳ㡪ࡀ࠶ࡿ࠿ ྰ࠿࡟ࡘ࠸࡚ゎᯒࡋࡓࠋඛయ཯ᛂࡣཷ⢭࡟࠾ࡅࡿᚲせ୙ྍḞ࡞ࣉࣟࢭࢫ࡛࠶ࡿࡢ࡛㸪Mg2+ ཬࡧ PMB ࡀ┤᥋⢭Ꮚᶵ⬟࡬ཬࡰࡍᙳ㡪ࢆㄪ࡭ࡓᚋ㸪୧⪅ࡢ෭ⶶಖᏑ⢭ᾮ࡬ࡢῧຍࡀಖᏑ ᚋࡢ⢭Ꮚࡢ୍⯡ᛶ≧ཬࡧඛయ཯ᛂ࡬ཬࡰࡍᙳ㡪ࢆゎᯒࡋ㸪୧⪅ࢆẚ㍑ࡋࡓࠋ  ➨ 2 ❶➨ 1 ⠇࡛ࡣ㸪Ca2+/A23187 ࡟ࡼࡾㄏ㉳ࡉࢀࡿඛయ཯ᛂ࡬ཬࡰࡍ Mg2+ ࡢᙳ㡪ࢆ ㄪ࡭ࡓࠋ0–3 mM ࡢ MgSO4 ࠶ࡿ࠸ࡣ MgCl2 ࡢᏑᅾୗ࡛㸪⢭Ꮚࢆ่⃭ࡋࡓᚋࡢඛయ཯ᛂㄏ ㉳⋡ࡣ㸪࠸ࡎࢀࡢ᫬㛫࡟࠾࠸࡚ࡶ௚ࡢ⃰ᗘ࡜ẚ࡭࡚ 0.1 mM ࡢ MgCl2 ࠶ࡿ࠸ࡣ MgSO4 ࡛᭷ព࡟పୗࡋࡓࠋ⢭Ꮚࢆ 0–3 mM MgSO4 ࡢᏑᅾୗ࡛ Ca2+/A23187 ࡟ࡼࡾ 60 ศ㛫ࡲ࡛ ่⃭ࡋࡓ⤖ᯝ㸪Ca2+/A23187 ࡟ࡼࡾㄏ㉳ࡉࢀࡓඛయ཯ᛂࡣ MgSO 4 ࡟ࡼࡾ⃰ᗘ౫Ꮡⓗ࡟ᢚ ไࡉࢀ㸪ࡑࡢᢚไࡣ 0.1 mM ࡛᭱኱࡛࠶ࡗࡓࠋ௨ୖࡼࡾ μM ࣞ࣋ࣝࡢ Mg2+ ࡟ࡼࡾ mM ࣞ࣋ࣝࡢ Ca2+ Ꮡᅾୗ࡛ Ca2+/A23187 ࡟ࡼࡾㄏ㉳ࡉࢀࡿඛయ཯ᛂࡣᢚไࡉࢀࡿࡇ࡜ࢆ♧ ࡋࡓࠋ  ➨ 2 ❶➨ 2 ⠇ࡢ◊✲ࡣ㸪ᕼ㔘ࡋࡓࣈࢱ⢭ᾮ࡟ Mg2+ ࢆῧຍࡋ࡚ಖᏑࡋࡓሙྜࡢಖᏑᚋ ࡢ⢭Ꮚࡢ㐠ືᛶཬࡧඛయ཯ᛂ࡬ཬࡰࡍᙳ㡪ࢆㄪ࡭ࡓࠋMgSO4 (0–8 mM) ࡢῧຍ࡛ࡣಖᏑᚋ ࡢ⢭Ꮚࡢ㐠ືᛶ࡟ᙳ㡪ࡋ࡞࠿ࡗࡓࡀ㸪ಖᏑࡋࡓ⢭Ꮚࡢඛయ཯ᛂࢆCa2+/A23187 ࡟ࡼࡾ่⃭ ࡋࡓ⤖ᯝ㸪่⃭ࡢ 10 ཬࡧ 15 ศ࡟࠾࠸࡚ࡑࢀࡒࢀ 4 mM ῧຍࡢ⢭Ꮚ࡟࠾࠸࡚௚ࡢ⃰ᗘ࡟ ẚ࡭᭷ព࡟ඛయ཯ᛂㄏ㉳⋡ࡀపୗࡋࡓࠋ่⃭⤊஢ᚋࡢ⢭Ꮚࡢ㐠ື⋡࡜⏕Ꮡ⋡ࡣ MgSO4 ῧ ຍ࡟ࡼࡿኚ໬ࡀぢࡽࢀ࡞࠿ࡗࡓࡇ࡜࠿ࡽ㸪ㄏ㉳ࡉࢀࡓඛయ཯ᛂࡣኚᛶ࡟ࡼࡿඛయࡢ႙ኻ࡛ ࡣ࡞࠸ࡇ࡜ࢆ☜ㄆࡋࡓࠋ௨ୖࡢ⤖ᯝࡣ㸪ኟᏘ୙ዷ⑕࡟≉ᚩⓗ࡛࠶ࡿ⢭Ꮚࡢ㐣๫࡞ඛయ཯ᛂ ㄏ㉳ࢆ⦆࿴࡛ࡁࡿ᪉ἲࡢ㛤Ⓨ࡟᭷⏝࡞᝟ሗࢆᥦ౪ࡋࡓࠋ  PMB ࡣయ⣽⬊࡟࠾࠸࡚ࡣࢩࢢࢼࣝኚ᥮ᶵᵓ࡟㛵୚ࡍࡿ཯ᛂ⤒㊰ࢆㄪ⠇ࡍࡿࡇ࡜ࡀ▱ࡽ ࢀ࡚࠸ࡿࡇ࡜࠿ࡽ㸪➨ 3 ❶➨ 1 ⠇ࡢᐇ㦂࡛ࡣ㸪PMB ࡀ┤᥋⢭Ꮚ࡬ཬࡰࡍᙳ㡪ࢆㄪ࡭ࡓࠋ 0–100 μM ࡢ PMB ࡢᏑᅾୗ࡛⢭Ꮚࢆᇵ㣴ࡋࡓ⤖ᯝ㸪㐠ື⋡࡜⏕Ꮡ⋡ࡣ㸪75 μM ཬࡧ 100 μM ࡢ PMB ῧຍ࡛᭷ព࡟పୗࡋࡓࠋ┤㐍㐠ື⋡ࡣ 25 μM ௨ୖࡢ㸪άⓎࡉࡣ 75 μM ௨ ୖࡢ PMB ῧຍ࡟ࡼࡾࡑࢀࡒࢀ᭷ព࡟పୗࡋࡓࡀ㸪จ㞟ࡣ࠸ࡎࢀࡢ⃰ᗘ࡟࠾࠸࡚ࡶᙳ㡪ࢆ ཷࡅ࡞࠿ࡗࡓࠋ୍᪉㸪㐠ື⋡࡜⏕Ꮡ⋡࡟ᙳ㡪ࡋ࡞࠿ࡗࡓ⃰ᗘ࡛࠶ࡿ 0–50 μM ࡢ PMB Ꮡ ᅾୗ࡛๓ᇵ㣴ࡋࡓᚋ㸪Ca2+/A23187 ࡟ࡼࡾඛయ཯ᛂࢆ่⃭ࡋࡓ⤖ᯝ㸪0.01–0.5 μM ཬࡧ 1– 50 μM ࡢῧຍ࡛ࡑࢀࡒࢀ่⃭ࡢ 10 ศཬࡧ 15 ศ࡛ㄏ㉳ࡉࢀࡓඛయ཯ᛂࡀቑᙉࡉࢀࡓࠋ  ➨ 3 ❶➨ 2 ⠇ࡢ◊✲࡛ࡣ㸪✀ࠎࡢ⃰ᗘ㸦0–75 μM㸧ࡢ PMB ࢆῧຍࡋ࡚ᕼ㔘ᚋಖᏑࡋ ࡓ⢭Ꮚࡢ㐠ືᛶࢆㄪ࡭ࡿ࡜ඹ࡟ PMB ῧຍࡀඛయ཯ᛂ࡬ཬࡰࡍᙳ㡪ࢆㄪ࡭ࡓࠋಖᏑࡋࡓ⢭ Ꮚࡢ㐠ື⋡ࡣ PMB ࡢῧຍ࡟ࡼࡗ࡚ࡣ᭷ព࡞ᙳ㡪ࢆཷࡅ࡞࠿ࡗࡓࡀ㸪┤㐍㐠ື⋡ࡀ PMB ࡢῧຍ⃰ᗘࡢୖ᪼࡜ඹ࡟ୖ᪼ࡋࡓࠋಖᏑࡋࡓ⢭ᏊࢆὙίᚋ㸪ྠᵝ࡟ 30 ศࡲ࡛ Ca2+/A23187 Ꮡᅾୗ࡛ᇵ㣴ࡋඛయ཯ᛂࢆ่⃭ࡋࡓࠋᇵ㣴⤊஢ᚋ㸪☜ㄆࡢ┠ⓗ࡛㸪⢭Ꮚࡢ㐠ື⋡㸪┤㐍㐠 ື⋡㸪⏕Ꮡ⋡㸪άⓎࡉཬࡧจ㞟ࢆㄪ࡭ࡓࠋᇵ㣴 15 ศཬࡧ 30 ศ࡟࠾࠸࡚ࡑࢀࡒࢀ 50 ཬ ࡧ 75 μM ࡢ PMB ῧຍ࡟ࡼࡾ Ca2+/A23187 ࡟ࡼࡾㄏ㉳ࡉࢀࡓඛయ཯ᛂࡀቑᙉࡉࢀࡓࠋ25 μM ࡢ PMB ࡢῧຍ࡟ࡼࡾ㐠ື⋡ཬࡧ⏕Ꮡ⋡ࡀ௚ࡢ⃰ᗘ࡟ẚ࡭࡚᭷ព࡟పୗࡋࡓࡀ㸪ࡇࢀ ࡽࡢ௨እࡢ⢭Ꮚࡢᛶ≧ࡣ PMB ῧຍ࡟ࡼࡾᙳ㡪ࢆཷࡅ࡞࠿ࡗࡓࠋ௨ୖࡼࡾ㸪ࣈࢱ⢭ᾮࢆ PMB Ꮡᅾୗ࡛ಖᏑࡍࡿࡇ࡜ࡀ࡛ࡁ㸪ࡇࢀ࡟ࡼࡾ㐠ື⋡࡜⏕Ꮡ⋡ࡣ୙ኚ࡛┤㐍㐠ື⋡ࡀୖ ᪼ࡋඛయ཯ᛂࡢㄏ㉳ࡀቑᙉࡉࢀࡿࡇ࡜ࢆ♧ࡋࡓࠋ  ௨ୖࡼࡾ㸪ᮏ◊✲ࡣ࣐ࢢࢿࢩ࣒࢘ࡣ㐠ືᛶࢆᦆ࡞࠺ࡇ࡜࡞ࡃࣈࢱ෭ⶶಖᏑ⢭ᾮ࡬ῧຍࡍ ࡿࡇ࡜ࡀ࡛ࡁ㸪ࡲࡓ㸪ῧຍ࡟ࡼࡾඛయ཯ᛂࡢㄏ㉳ࡀᢚไࡉࢀࡿࡇ࡜࠿ࡽ㐣๫࡞ඛయ཯ᛂࡢ ㄏ㉳ࢆ⦆࿴ࡋ㸪ኟᏘ࡟᥇ྲྀࡉࢀࡿ⢭Ꮚࡢཷ⢭⬟ຊࢆ㧗ࡵࡽࢀࡿྍ⬟ᛶࢆᣦ᦬ࡋࡓࠋ୍᪉㸪 PMB ࢆಖᏑ⢭ᾮ࡬ῧຍࡍࡿࡇ࡜࡟ࡼࡾ┤㐍㐠ື⋡ࢆ㧗ࡵࡿࡇ࡜ࡀ࡛ࡁ㸪ྠ᫬࡟༸Ꮚ࡬ࡢ ౵ධ࡟ᚲせ୙ྍḞ࡞ඛయ཯ᛂㄏ㉳⬟ຊࡀ㧗ࡵࡽࢀ㸪ཷ⢭⬟ຊࢆୖ᪼ࡉࡏᚓࡿྍ⬟ᛶࢆ♧ࡋ

(5)

ࡓࡀ㸪ኟᏘ࡟᥇ྲྀࡉࢀࡿ⢭Ꮚ࡬ῧຍࡍࡿࡇ࡜࡟ࡼࡾ㸪㐣๫࡟ㄏ㉳ࡉࢀࡿඛయ཯ᛂࢆࡉࡽ࡟ ቑᙉࡍࡿྍ⬟ᛶࡀ࠶ࡾ㸪ኟᏘ࡟౑⏝ࡍࡿ㝿࡟ࡣ⪃៖ࡀᚲせ࡛࠶ࡿࡇ࡜ࢆᣦ᦬ࡋࡓࠋ  ௨ୖ࡟ࡘ࠸࡚㸪ᑂᰝጤဨ඲ဨ୍⮴࡛ᮏㄽᩥࡀᒱ㜧኱Ꮫ኱Ꮫ㝔㐃ྜ⋇་Ꮫ◊✲⛉ࡢᏛ఩ㄽ ᩥ࡜ࡋ࡚༑ศ౯್ࡀ࠶ࡿ࡜ㄆࡵࡓࠋ  ᇶ♏࡜࡞ࡿᏛ⾡ㄽᩥ

㸯㸧㢟 ┠㸸 Polymyxin B enhances acrosomal exocytosis triggered by calcium and the calcium ionophore A23187 in ejaculated boar spermatozoa

 ⴭ ⪅ ྡ㸸Akter, Q. S., Rajabi-Toustani, R., Shimizu, K., Kuwahara, Y. and Murase, T.  Ꮫ⾡㞧ㄅྡ㸸Animal Science Journal

 ᕳ࣭ྕ࣭㡫࣭Ⓨ⾜ᖺ㸸In Press

᪤Ⓨ⾲Ꮫ⾡ㄽᩥ

㸯㸧㢟 ┠㸸 Effect of calmodulin on the stimulation of capacitation and acrosome reaction of frozen thawed bull spermatozoa

  ⴭ ⪅ ྡ㸸Akter, Q. S., Tareq, K. M. A., Hamano, K. and Gilchrist, R. B.   Ꮫ⾡㞧ㄅྡ㸸Bangladesh Journal of Animal Science

  ᕳ࣭ྕ࣭㡫࣭Ⓨ⾜ᖺ㸸45㸦3㸧㸸1-9㸪2016 

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