Title
Studies on the Relationship among Standard Parameters, Acrosomal Integrity, and the In Vitro Inducibility of
Hyperactivation and the Acrosome Reaction in Frozen-thawed Bull Spermatozoa( 内容と審査の要旨(Summary) )
Author(s) Reza Rajabi-Toustani
Report No.(Doctoral Degree) 博士(獣医学) 甲第538号 Issue Date 2019-03-13 Type 博士論文 Version ETD URL http://hdl.handle.net/20.500.12099/77969 ※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。
Ặྡ㸦ᮏ㸦ᅜ㸧⡠㸧 5H]D5DMDEL7RXVWDQL㸦࣭ࣛࣥࢫ࣒ࣛඹᅜ㸧 ᣦ ᑟ ᩍ ဨ Ặ ྡ ᒱ㜧Ꮫ ᩍᤵ ᮧ ℩ ဴ ☻ Ꮫ ࡢ ✀ 㢮 ༤ኈ㸦⋇་Ꮫ㸧 Ꮫ グ ␒ ྕ ⋇་༤⏥➨㸳㸱㸶ྕ 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 ◊✲ᣦᑟࢆཷࡅࡓᏛ ᒱ㜧Ꮫ Ꮫ ㄽ ᩥ 㢟 ┠ 6WXGLHVRQWKH5HODWLRQVKLSDPRQJ6WDQGDUG3DUDPHWHUV $FURVRPDO,QWHJULW\DQGWKH,Q9LWUR,QGXFLELOLW\RI +\SHUDFWLYDWLRQDQGWKH$FURVRPH5HDFWLRQLQ )UR]HQWKDZHG%XOO6SHUPDWR]RD 㸦࢘ࢩ⤖⼥ゎ⢭Ꮚ࠾ࡅࡿ୍⯡ᛶ≧㸪ඛయṇᖖᛶཬࡧ ࣁࣃ࣮ࢡࢳ࣮࣋ࢩࣙࣥඛయᛂࡢయእㄏ㉳⬟ຊ ࡢ㛫࠾ࡅࡿ┦㛵ಀ㛵ࡍࡿ◊✲㸧 ᑂ ᰝ ጤ ဨ ᰝ ᒱ 㜧 Ꮫ ᩍ ᤵ ⊦ ᓥ ᗣ 㞝 ᰝ ᖏᗈ␆⏘Ꮫ ᩍ ᤵ ༡ ಖ Ὀ 㞝 ᰝ ᒾ ᡭ Ꮫ ᩍ ᤵ బ ⸨ ⦾ ᰝ ᮾி㎰ᕤᏛ ᩍ ᤵ ⏣ ୰ ▱ ᕫ ᰝ ᒱ 㜧 Ꮫ ᩍ ᤵ ᮧ ℩ ဴ ☻ Ꮫㄽᩥࡢෆᐜࡢせ᪨ ∵ࡢ⦾Ṫ࠾࠸࡚ࡣ⤖⢭ᾮࡼࡿேᕤᤵ⢭㸦AI㸧ࡀ᭱ࡶ㔜せ࡞ᢏ⾡ࡢ୍ࡘ࡛࠶ࡿࠋAI ⏝ࡍࡿ⢭ᾮࡣ᳨ᰝࡉࢀ୍ᐃ௨ୖࡢᛶ≧ࡢ⢭ᾮࡀ⤖ಖᏑࡉࢀࡿࠋࡋࡋ㸪୍⯡᳨ᰝ࡛ ࡣṇᖖ࡛࠶ࡿࡶࢃࡽࡎ㸪ேᕤᤵ⢭ᚋࡢཷ⫾⋡ࡀపୗࡍࡿ࠸ࢃࡺࡿప⦾Ṫࡢಶయࡀ Ꮡᅾࡍࡿࠋᮏㄽᩥࡢ◊✲ࡣ㸪⢭ᾮࡢ୍⯡᳨ᰝ࡛ࡣ᳨ฟ࡛ࡁ࡞࠸ప⦾Ṫࡢ✀㞝∵ࢆ᳨ฟ࡛ ࡁࡿ᪂つ᳨ᰝἲࢆ㛤Ⓨࡍࡿࡓࡵࡢᇶ♏▱ぢࢆᚓࡿࡇࢆ┠ⓗࡋ㸪㯮ẟ✀✀㞝∵ࡢ⤖ ⼥ ゎ ⢭ ᾮ ࢆ ⏝ ࠸ ࡚ 㸪 ୍ ⯡ ⢭ ᾮ ᛶ ≧ 3 ✀ ࡢ ᪂ つ ᳨ ᰝ ἲ ࡋ ࡚ Fluorescein isothiocyanate-conjugated peanut agglutinin (FITC-PNA) ⺯ගⰍ⣲ᰁⰍࡼࡾホ౯ࡋࡓ⢭Ꮚࡢ
ඛయṇᖖᛶ㸪࢝ࣝࢩ࣒࢘࢜ࣀ࣍ A23187 ࡼࡾㄏ㉳ࡋࡓ⢭Ꮚ㢌㒊ࡢඛయᛂ㸦AR㸧 ཬࡧ⣽⬊㏱㐣ᆺ cAMP ࢼࣟࢢ, sp-5,6-dichloro-1-β-D-ribofuranosylbenzimida- zole-3',5'-monophosphorothioate (cBiMPS), ࡼࡾㄏ㉳ࡋࡓ㠴ẟࡢࣁࣃ࣮ࢡࢳ࣮࣋ࢩࣙ ࣥ㸦HA㸧ࡢ㛫࠾ࡅࡿ┦㛵ಀࢆゎᯒࡋࡓࠋ ➨ 2 ❶࡛ࡣ㸪 FITC-PNA ⺯ගⰍ⣲ᰁⰍࡢᰁⰍ᮲௳ࡢᨵⰋࢆ⾜ࡗࡓࠋᮏᰁⰍ࡛ࡣ㸪ᅛᐃ ࡋࡓ⢭ᏊⰍ⣲ࢆ⣽⬊ෆ㏱㐣ࡉࡏࡿ┠ⓗ࡛ Triton X-100 ࡼࡿ㏱㐣ฎ⌮ࢆ⾜ࡗࡓᚋᰁ Ⰽࢆ⾜࠺ࡀ㸪ࡇࡢ㐣⛬࡛ேⅭⓗ࡞ࢲ࣓࣮ࢪࡀᑡ࡞ࡃ㸪ඛయṇᖖᛶࢆࡼࡾṇ☜ホ౯࡛ࡁࡿ ᰁⰍ᮲௳ࢆ᳨ウࡋࡓࠋᐇ㦂1 ࡼࡾ㸪⢭ᏊࢆᰁⰍࡍࡿሙྜሬᶆᮏୖ࡛ᰁⰍࡍࡿࡼࡾࡶ⢭Ꮚ ࡀᾋ㐟ࡋࡓ≧ែ࡛ᰁⰍ㸦ᾋ㐟ἲ㸧ࡋࡓ᪉ࡀࡼࡃ㸪ࡑࡢሙྜㄪࡓ3 ✀ࡢ㏥Ⰽ㜵Ṇ, DABCO,
SlowFade Diamond® (SlowFade) ཬࡧ ProLong Diamond® (ProLong) ࡢ୰࡛ SlowFade ࡀ㐺ࡋ 㸦㸰㸯㸧
࡚࠸ࡿࡀ㸪ಖᏑࡣ᥎ዡࡉࢀ࡞࠸ࡇ㸪ࡇࡢሙྜࡀ୍ࢧࣥࣉࣝࢆಖᏑࡍࡿሙྜࡣ㏥Ⰽ㜵Ṇ ࡋ࡚ࡣ ProLong ࡀ᭱㐺࡛࠶ࡿࡇ㸪ࡀ୍ሬᶆᮏࢆಖᏑࡍࡿሙྜࡣ㏥Ⰽ㜵Ṇࡋ ࡚ DABCO ࡣ㑊ࡅࡿࡁ࡛࠶ࡿࡇࡀ♧ࡉࢀࡓࠋᐇ㦂 2 ࡛ࡣᾋ㐟ἲࢆ⏝࠸࡚㸪Triton X-100 ࡢ⃰ᗘࢆ᳨ウࡋࡓ⤖ᯝ㸪㏻ᖖ⏝࠸ࡽࢀࡿ 1% ࡽ 0.1% ࡲ࡛ῶᑡ࡛ࡁࡿࡇࡀ᫂ࡽ࡞ ࡗࡓࠋᐇ㦂 3 ཬࡧ 4 ࡽ㸪᭱ࡶ⢭Ꮚࢲ࣓࣮ࢪࡀᑡ࡞࠸ᅛᐃ᮲௳ࡣ 2% ࡛ 30 ศ㛫࡛࠶ ࡗࡓࠋ ➨ 3 ❶࡛ࡣ㸪✀ࠎࡢ␗࡞ࡿ⤖⢭ᾮࢧࣥࣉࣝ࠾࠸࡚ A23187 ࡼࡾㄏ㉳ࡋࡓඛయ ᛂࡢ⤒ኚࢆㄪࡓᚋ㸪⢭Ꮚࡢᚑ᮶ࡢᣦᶆ㸪FITC-PNA ᰁⰍࡼࡿඛయṇᖖᛶཬࡧඛయ ᛂㄏ㉳⋡ࡢ⤒ኚࣃࢱ࣮ࣥࡢ㛫ࡢ┦㛵ಀࢆゎᯒࡋࡓࠋࡑࡢ⤖ᯝ㸪ඛయᛂㄏ㉳⋡ ࡢ⤒ኚࡣᛂ࣮࢝ࣈࡢഴᩳࡽ 3 ࢢ࣮ࣝࣉ㸦AR ࢢ࣮ࣝࣉ㸧ศ㢮࡛ࡁࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋࡍ࡞ࢃࡕ㸪⦆ࡸ࡞ୖ᪼ࢆ♧ࡍS Ꮠ≧࣮࢝ࣈ㸦Group 1㸧㸪่⃭┤ᚋࡽ⦆ࡸ ୖ᪼ࡍࡿ࣮࢝ࣈ㸦Group 2㸧ཬࡧ่⃭┤ᚋࡽᛴ⃭ୖ᪼ࡍࡿ࣮࢝ࣈ㸦Group 3㸧࡛࠶ ࡗࡓࠋḟ࠸࡛㸪AR ࢢ࣮ࣝࣉ㛫࡛ᚑ᮶ࡢᣦᶆཬࡧ FITC-PNA ᰁⰍࡼࡿඛయṇᖖᛶࢆẚ㍑ ࡋࡓ⤖ᯝ㸪࠸ࡎࢀࡢᣦᶆࡶ᭷ពᕪࡀㄆࡵࡽࢀ࡞ࡗࡓࠋ୍᪉㸪FITC-PNA ᰁⰍࡼࡿඛయ ṇᖖᛶᚑ᮶ࡢᣦᶆࢆẚ㍑ࡋࡓ⤖ᯝ㸪࠸ࡎࢀࡢᣦᶆࡶඛయṇᖖᛶ᭷ព࡞┦㛵ࢆ♧ࡋࡓࠋ ࡞࠾㸪⏝࠸ࡓࢧࣥࣉࣝࡢ㛫࡛ࡣ㸪᥇⢭ࡢಶయࡢᖺ㱋ᚑ᮶ࡢᣦᶆ㸪ඛయṇᖖ⋡ཬࡧ AR ࢢ ࣮ࣝࣉࡢ㛫㛵㐃ࡣぢฟࡉࢀ࡞ࡗࡓࠋࡲࡓ㸪୍⯡ᛶ≧ࡀṇᖖ࡛࠶ࡿࡶࢃࡽࡎཷ ⫾⋡ࡀప࠸ࡇࡀࢃࡗ࡚࠸ࡓಶయ2 㢌㐠ື⋡ࡀపୗࡋࡓࡀཷ⫾ྍ⬟࡛࠶ࡗࡓಶయ 1 㢌 ࡢ AR ㄏ㉳ࣃࢱ࣮ࣥࡣ㸪࠸ࡎࢀࡢಶయࡶ Group 3 ศ㢮ࡉࢀࡓࠋ௨ୖࡼࡾ㸪AR ࡢᛂ ᛶࡼࡾ㸪⢭Ꮚࡢᶵ⬟≧ែࢆศ㢮ࡍࡿࡇࡀྍ⬟࡛࠶ࡾ㸪ࡇࢀࡣᚑ᮶ࡢᣦᶆཬࡧඛయṇᖖ ᛶࡣ⊂❧࡛࠶ࡿࡇࡀ♧၀ࡉࢀࡓࠋࡲࡓ㸪ඛయṇᖖᛶ AR ᛂᛶࡢ⤌ࡳྜࢃࡏࡀప⦾ Ṫಶయࢆ᳨ฟࡍࡿࡓࡵࡢ⢭Ꮚṇᖖ㛵ࡍࡿ㔜せ࡞ሗ࡞ࡾᚓࡿࡇࡀᮇᚅࡉࢀࡿࠋ ➨ 4 ❶࡛ࡣ㸪⢭Ꮚࡢ୍⯡ᛶ≧㸪FITC-PNA ᰁⰍࡼࡿඛయṇᖖᛶ㸪cBiMPS ࡼࡾㄏ ㉳ࡋࡓ HA ཬࡧ A23187 ࡼࡾㄏ㉳ࡋࡓඛయᛂࡢ㛫࠾ࡅࡿ㛵㐃ࢆゎᯒࡋࡓࠋࡑࡢ⤖ ᯝ㸪㐠ື⋡ࡀప࠸ಶయ A B ཬࡧ㐠ື⋡ࡀṇᖖ࡛࠶ࡿࡀపཷ⫾࡛࠶ࡿࡇࡀ᪤▱ࡢಶయ C ࡛ࡣඛయṇᖖᛶࡀపࡃ㸪HA ㄏ㉳⋡ࡣ 0% ࠶ࡿ࠸ࡣప⋡࡛࠶ࡗࡓࠋ୍᪉㸪㐠ື⋡ࡀṇᖖ ࡛㧗࠸ಶయ D ཬࡧ E ࡛ࡣ㸪ඛయṇᖖ⋡ࡀ㧗ࡃ HA ࡣ㧗⋡ㄏ㉳ࡉࢀ㸪HA ㄏ㉳⬟ຊඛ యṇᖖ⋡ࡣẚ㍑ⓗࡼࡃ୍⮴ࡋ࡚࠸ࡓࠋࡲࡓ㸪㐠ື⋡ࡀపࡗࡓಶయ A ཬࡧప⦾Ṫࡢಶ య C ࡢ AR ࢢ࣮ࣝࣉࡣ Group 3 㸪ಶయ B ࡣ Group 2 㸪㐠ື⋡ࡀṇᖖ࡞ಶయ D ཬ ࡧ E ࡣ Group ࠸ࡎࢀࡶ Group 2 ศ㢮ࡉࢀࡓࠋ௨ୖࡼࡾ㸪㐠ື⋡ࡣ࠶ࡿ⛬ᗘ HA ㄏ㉳ ⬟ຊ㛵㐃ࡍࡿࡀ㸪ಶయ C ࡀ♧ࡍࡼ࠺㐠ື⋡ࡀṇᖖ࡛࠶ࡗ࡚ࡶ HA ㄏ㉳⬟ຊࡀప࠸ಶ యࡢᏑᅾࡍࡿࡇࡇࡀ⪃࠼ࡽࢀࡓࠋࡲࡓ㸪㐠ື⋡ HA ㄏ㉳⋡ࡀṇᖖ࡞ಶయ D ཬࡧ E ࡣ AR ᛂࣃࢱ࣮ࣥࡢ Group 2 ศ㢮ࡉࢀࡓࡇཬࡧ➨ 2 ࡢᐇ㦂࡛ప⦾Ṫࡢಶయࡣ Group 3 ศ㢮ࡉࢀࡓࡇࡽ㸪Group 2 ࡣṇᖖಶయࡀศ㢮ࡉࢀࡿࡇࡀ⪃࠼ࡽࢀࡓࠋࡋ ࡋ㸪ಶయ B ࡣ㐠ື⋡ HA ㄏ㉳⋡ࡀప࠸ࡶࢃࡽࡎ AR ࢢ࣮ࣝࣉࡣ Group 2 ศ㢮ࡉࢀࡓࡇࡽ㸪HA ㄏ㉳⬟ຊ AR ᛂࣃࢱ࣮ࣥࡣ␗࡞ࡿ⢭Ꮚࡢᶵ⬟ࢆ᳨ฟࡋ࡚࠸ ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ ௨ୖࡢ⤖ᯝࡣ㸪FITC-PNA ᰁⰍࡼࡿඛయṇᖖᛶ㸪HA AR ㄏ㉳⬟ຊࡣࡑࢀࡒࢀ⢭Ꮚ ࡢ␗࡞ࡿᶵ⬟ࢆᫎࡋ࡚࠸ࡿࡇࡀ♧၀ࡉࢀࡓࠋࡇࢀࡽࡣ୍⯡ᛶ≧ࡀṇᖖ࡛࠶ࡿࡶ ࢃࡽࡎప⦾Ṫࢆ♧ࡍಶయࢆ᳨ฟࡍࡿ᪂つ᳨ᰝἲࡢ㛤Ⓨࡢᇶ♏▱ぢ࡞ࡿࡇࡀᮇᚅࡉ ࢀࡿࠋ ᑂ ᰝ ⤖ ᯝ ࡢ せ ᪨ ᮏㄽᩥࡣ㸪㯮ẟ✀࠾࠸࡚㸪⢭ᾮࡢ୍⯡᳨ᰝ࡛ࡣ᳨ฟ࡛ࡁ࡞࠸ప⦾Ṫࡢ✀㞝∵ࢆ᳨
ฟ࡛ࡁࡿ᪂つ᳨ᰝἲࢆ㛤Ⓨࡍࡿࡓࡵࡢᇶ♏▱ぢࢆᚓࡿࡇࢆ┠ⓗࡋ㸪⤖⼥ゎ⢭ᾮࢆ⏝ ࠸୍࡚⯡⢭ᾮᛶ≧ 3 ✀ࡢ᪂つ᳨ᰝἲࡋ࡚⢭Ꮚࡢඛయṇᖖᛶ㸪యእ࠾ࡅࡿ⢭Ꮚ㢌㒊ࡢ ඛయᛂ㸦AR㸧ཬࡧ⢭Ꮚᑿ㒊ࡢࣁࣃ࣮ࢡࢳ࣮࣋ࢩࣙࣥ㸦HA㸧ㄏ㉳⬟ຊࡢ㛫࠾ࡅ ࡿ┦㛵ಀࢆゎᯒࡋࡓࠋ
➨1 㸪⢭Ꮚࡢඛయṇᖖᛶࢆホ౯ࡍࡿ FITC ᶆ㆑-Peanut จ㞟⣲㸦FITC-PNA㸧⺯ගⰍ⣲
ᰁⰍࡢᰁⰍ᮲௳ࡢᨵⰋࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝ㸪⢭Ꮚࡀᾋ㐟≧ែࡢ᪉ࡀ㸪ሬࡋࡓ≧ែ࡛ᰁⰍ ࡍࡿ᪉ἲẚ࡚⢭ᏊࡢேⅭⓗࢲ࣓࣮ࢪࡀᑡ࡞࠸ࡇࢆ᫂ࡽࡋࡓࠋࡲࡓ㸪ᨵⰋࡋࡓ
᪂つ᪉ἲࡢᰁⰍ᮲௳ࡋ࡚㸪ᅛᐃࡣ 2% ࣃ࣒ࣛ࣍ࣝࣝࢹࣄࢻ ࡛ 30 ศ㛫, ㏱㐣ฎ⌮ࡣ
0.1% Triton ࡚ 5 ศ㛫㸪㏥Ⰽ㜵Ṇࡋ࡚ᕷ㈍ࡢ SlowFade® Diamond ࢆ⏝࠸ࡿ㸪㏥Ⰽ㜵 Ṇࢆຍ࠼ࡓᚋࡣಖᏑࡏࡎ┤ࡕほᐹࡍࡿࡇ㸪ࢆᥦࡋࡓࠋ ➨2 㸪✀ࠎࡢ␗࡞ࡿ⤖⢭ᾮࢧࣥࣉࣝ࠾࠸࡚࢝ࣝࢩ࣒࢘࢜ࣀ࣍ A23187 ࡼࡾ ㄏ㉳ࡋࡓඛయᛂࡢ⤒ኚࢆㄪࡓᚋ㸪⢭Ꮚࡢᚑ᮶ࡢᣦᶆ㸪FITC-PNA ᰁⰍࡼࡿඛయ ṇᖖᛶཬࡧඛయᛂㄏ㉳⋡ࡢ⤒ኚࣃࢱ࣮ࣥࡢ㛫ࡢ┦㛵ಀࢆゎᯒࡋࡓࠋࡑࡢ⤖ᯝ㸪 ᛂ࣮࢝ࣈࡢഴᩳࡽඛయᛂㄏ㉳⋡ࡢ⤒ኚࡣ 3 ࢢ࣮ࣝࣉ㸦AR ࢢ࣮ࣝࣉ㸧ศ㢮࡛ ࡁࡿࡇࢆ᫂ࡽࡋࡓࠋAR ࢢ࣮ࣝࣉ㛫࡛ᚑ᮶ࡢ⢭Ꮚᛶ≧ࡢᣦᶆཬࡧ FITC-PNA ᰁⰍ ࡼࡿඛయṇᖖᛶࢆẚ㍑ࡋࡓ⤖ᯝ㸪࠸ࡎࢀࡢᣦᶆࡶ᭷ពᕪࡀㄆࡵࡽࢀ࡞ࡗࡓࠋ୍᪉㸪 FITC-PNA ᰁⰍࡼࡿඛయṇᖖᛶᚑ᮶ࡢᣦᶆࢆẚ㍑ࡋࡓ⤖ᯝ㸪࠸ࡎࢀࡢᣦᶆࡶඛయṇ ᖖᛶࡢ㛫᭷ព࡞┦㛵ࢆぢฟࡋࡓࠋ௨ୖࡼࡾ㸪ඛయṇᖖᛶࡣ୍⯡ᛶ≧┦㛵ࡍࡿࡀ㸪AR ࡢㄏ㉳⬟ຊࡣᚑ᮶ࡢᣦᶆཬࡧඛయṇᖖᛶࡣ⊂❧࡛࠶ࡿࡇࢆ♧ࡋࡓࠋ ➨3 ࡢᐇ㦂࡛ࡣ㸪⢭Ꮚࡢ୍⯡ᛶ≧㸪FITC-PNA ᰁⰍࡼࡿඛయṇᖖᛶ㸪cAMP ࢼࣟࢢ cBiMPS ࡼࡾㄏ㉳ࡋࡓ HA ཬࡧ A23187 ࡼࡾㄏ㉳ࡋࡓඛయᛂࡢ㛫࠾ࡅࡿ㛵㐃ࢆ ゎᯒࡋࡓࠋࡑࡢ⤖ᯝ㸪㐠ື⋡ࡣ࠶ࡿ⛬ᗘ HA ㄏ㉳⬟ຊ㛵㐃ࡍࡿࡀ㸪㐠ື⋡ࡀṇᖖ࡛࠶ࡗ ࡚ࡶ HA ㄏ㉳⬟ຊࡀప࠸ಶయࡢᏑᅾࡍࡿྍ⬟ᛶࢆᣦࡋࡓࠋࡲࡓ㸪HA ㄏ㉳⬟ຊ AR ࣃ ࢱ࣮ࣥࡣ␗࡞ࡿ⢭Ꮚࡢᶵ⬟ࢆ᳨ฟࡋ࡚࠸ࡿྍ⬟ᛶࢆᣦࡋࡓࠋ ௨ୖࡢ⤖ᯝࡽ㸪ᮏㄽᩥࡣඛయṇᖖᛶ㸪HA AR ㄏ㉳⬟ຊࡣࡑࢀࡒࢀ⢭Ꮚࡢ␗࡞ࡿᶵ ⬟ࢆᫎࡋ࡚࠸ࡿࡇࡀ♧၀ࡉࢀ㸪ࡇࢀࡽࡣ୍⯡ᛶ≧ࡀṇᖖ࡛࠶ࡿࡶࢃࡽࡎప⦾Ṫ ࢆ♧ࡍಶయࢆ᳨ฟࡍࡿ᪂つ᳨ᰝἲࡢ㛤Ⓨࡢ㈗㔜࡞ᇶ♏▱ぢࢆᥦ౪ࡋ࡚࠸ࡿุ᩿ࡋࡓࠋ ௨ୖࡘ࠸࡚㸪ᑂᰝጤဨဨ୍⮴࡛ᮏㄽᩥࡀᒱ㜧ᏛᏛ㝔㐃ྜ⋇་Ꮫ◊✲⛉ࡢᏛㄽ ᩥࡋ࡚༑ศ౯್ࡀ࠶ࡿㄆࡵࡓࠋ ᇶ♏࡞ࡿᏛ⾡ㄽᩥ
㸯㸧㢟 ┠㸸Methodological improvement of fluorescein isothiocyanate peanut agglutinin (FITC-PNA) acrosomal integrity staining for frozen-thawed Japanese Black bull spermatozoa
ⴭ ⪅ ྡ㸸 Rajabi-Toustani, R., Akter, Q. S., Almadaly, E. A., Hoshino, Y., Adachi, H., Mukoujima, K. and Murase, T.
Ꮫ⾡㞧ㄅྡ㸸The Journal of Veterinary Medical Science ᕳ࣭ྕ࣭㡫࣭Ⓨ⾜ᖺ㸸In Press
