ウズラの輸卵管における精子貯蔵に関する生理学的研究
全文
(2) # ". &. .,-/. ' (%
(3) $ *) +. !.
(4)
(5) . .
(6) }. ². 1. B f´. 3. G ]¶ªw]»RlPZ] X_]¶ª
(7) F¿|{ 9. B ¤´. 10. G vsUt (1). `ÄR. (2). ±¬. (3). UVJ nN ³q. (4). ] pV»R)$. (5). =¼?6,;3A. (6). ŹK?6,;3AÆHPLCÇ. (7). OV?6,;3AÆPLCÇ«d?6,;3A ÆTLCÇ. (8). NMR. (9). SST F¿_À. (10) SST TÁ (11) SST F¿¨ _ (12) RT-PCR (13) in situ 04<(%A. :@. (14) RNA-Seq cDNA ;4;< { (15) RNA-Seqde novo $@4<+3>@. 9=¯x. (16) ¥J¿ zN (17) 7,@-<k _ (18) ¢°¯x 20. C y (1). =¼?6,;3A
(8) UVJ nN OÁ. (2). HPLC
(9) UVJ nN OÁ. (3). PLC
(10) UVJ nN OÁ i.
(11) (4). NMR
(12) ]»RlPZ] {º_. (5). SST
(13) F¿ _À. (6). TÁ SST F¿¨. (7). SST
(14) MCT ¾I] c[. (8). RNA-Seq SST D UVJ ©D ¾I]e¯x. (9). UVJ
(15) ¿L¡. (10) SST D§ 7,@-<k Y ¦a. 25. H bo. 28. C F¿
(16) ] »RlP|{ B ¤´. 29. G vsUt. 31. (1). `ÄR. (2). ±¬. (3). ] pV»R)$. (4). ] §\ pH §M pH. (5). SST ¶ª] §M pH _. (6). ](/@ ATPase k _. (7). ATP _À. (8). ¢°¯x 35. C y (1). F¿] »R
(17) U iÃ. (2). u|¿ Q
(18) §\ pH ¿kS] »R
(19) U iÃ. (3). SST ¶ª] §M pH. (4). #;](/@ ATPase k. (5). <6. @1.!@¿@](/@. ATPase k
(20) U ià (6). F¿ Q] ATP À
(21) U iÃ. Y ¦a. 37. H bo. 40. ii.
(22) Y #;] »RPj §M. .=I½·. B ¤´. 41. G vsUt. 43. (1). `ÄR. (2). ±¬mK. (3). ] pV»R)$. (4). ] §M pH _. (5). ] §M Ca2+g _. (6). ] 7,@-<k _. (7). cAMP _À. (8). ATP _À. (9). ](/@ nN(/@ ATPase k _. (10) ]WS ³® (11) '@2_À (12) SDS-5<<=7-=Â~RÆSDS-PAGEÇ (13) "'@4?)*@ (14) ¢°¯x 51. C y (1). BisII BisV ] »R
(23) EiÃ. (2). BisII ] §
(24) EiÃ. (3). PKC
(25) ]'@2 <@¿S. (4). ]
(26) ^[ PKC &3A8 X_. Y ¦a. 53. H bo. 57. H £o. 58. µ¸. 64. hr. 65. iii.
(27) ʋ̻̣. AM:. acetoxymethyl. ATP:. adenosine 5′-triphosphate. ATPase: adenosine triphosphatase BCA:. bicinchoninic acid. BCECF: 2′, 7′-bis (carboxyethyl)-4 or 5- carboxyfluorescein BPB:. bromophenol blue. BW:. body weight. CoA:. coenzyme A. DAPI:. 4′,6-diamidino-2-phenylindole. DNA:. deoxyribonucleic acid. DNase I: deoxyribonuclease I DTT:. dithiothreitol. EIA:. enzyme immunoassay. FI:. fluorescent intensity. HEPES:. 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid. HMBC:. hetero-nuclear multiple-bond coherence. HMQC:. hetero-nuclear multiple-quantum coherence. HRP:. horseradish peroxidase. IC50:. half maximal inhibitory concentration. IgG:. immunoglobulin G. MCT:. monocarboxylate transporter. NADH:. reduced nicotinamide adenine dinucleotide. 1.
(28) NMR:. nuclear magnetic resonance. OCT:. optimal cutting temperature. PCR:. polymerase chain reaction. PVDF:. polyvinylidene difluoride. QV:. quality value. RNA-Seq: RNA sequencing RNA:. ribonucleic acid. RT-PCR: reverse transcription PCR SDS:. sodium dodecyl sulfate. TCA:. tricarboxylic acid. Tris:. tris(hydroxymethyl)aminomethane. cAMP:. cyclic adenosine 3′,5′-monophosphate. cDNA:. complementary DNA. dATP:. deoxyadenosine triphosphate. mRNA:. messenger RNA. rRNA:. ribosomal RNA. 2.
(29) ˁʾ Ơ̿. ċɵ7Ģˏ50
(30) ˏŧ2ę7ͷ͟LǶΎʺΎʔ5þƲ%H. 2Lʚʔ2#0śȠ. 4ʁȬǁʋͧď#0+ RtD7E4ÈŚĢˏċɵ18
(31) Ǧę
(32) Ǧˏ7jQ \8̉%H7̋ʔ1
(33) ˏŧ8ęʅȋ7ďũ͐ďƹɵ͍7ɦƢčͻLƽʢ#
(34) ę5ż%H͐ďƹͪċL̞. 2ʢFI0H ĭǶ5Ǧõ"IHͻÜŧ2ˏŧ7͐. ďƹ8ÈŚĢˏċɵ7ĢˏLEGǀć"'D%%H. 25Ƭ͊.0HνNishigaki. et al., 2004; Shiba et al., 2008ξ ϧάLıAÈìĢˏċɵ50C
(35) ˏŧ2ęLĭǶ 5Ģˏ7Œ5ýʠ"'H. 2ĢˏLǀć"'D%%H 7+B5
(36) Ve8ǖę5. äʽ-
(37) H8ǖę¤5±ƂL̞
(38) e7ęˉ5ˏŧL͘͞%H RaZDv_7 E47ʵ18
(39) ±Ƃ̆ÈǖęL̺ʑ%HqX24HνDal Bosco et al., 2011; Brown, 2011ξ
(40) ś7ϧά18
(41) ±Ƃ8ǖę7jQ\5ƫΤL 4 IF7ʵ508
(42) e8ęĢˏĦ˻4ɷƾ54H?17Ύ
(43) ęˉ5ˏŧL͈̖ %H. 21H. 7ɽ͆8̋ʔ5͈ˏ2ķ9I
(44) Dz̚Dζά
(45) £ʁά
(46) ɱ̚ά
(47). ηά43
(48) Ļ§ά¼Ś7ċɵ1CƟ̫FIHɽ͆1HνBirikhead, 1993; Birkhead and Møller, 1993; Orr and Zuk, 2012; Suarez, 2008ξ #+.0
(49) ΛƹʁȬͭì17͈ˏ8
(50) ÈìĢˏċɵ508ˏŧ2ę7ʝÊʃͲ÷4ǶΎ2Œǂ1̞KIH. 2LÔ. ̶%H+B7̋ʔ4ǁʋ1H2HνHolt and Lloyd, 2010; Holt, 2011ξ ͈ˏ7ȀC̩4Ƽ˲8
(51) ˏŧLͲ÷4ǶΎ?1͈̖%H \LȀͲď%H. 25EGĢˏ7jQ. 21H Í9ʫÀƹ7xlʚνe.g. Oʱ
(52) nylʱ
(53) ef. ylʱξ18
(54) ǀɮ#+ôŠŠɼ8˫ȬȂ5ǩĐ7Ve2±ƂL̞
(55) )7Ư8Ƣ C±Ƃ%H. 24Ɠ7¤1ēƝ¼5CK+GĢˏęLʂ@ˢH 7E4Dz̚. 7ΛƹʁȬľ58ĢˏĿνspermathecaξ2ķ9IHľŮŨň#0G
(56) ĢˏĿ5͈B FI+ʁö7ˏŧLƁ#&/Ǧõ%H. 25E.0ĢˏLǀʽ"'Hνden Boer et al.,. 3.
(57) 2009ξ %4K-
(58) ʫÀƹ7xlʚ508
(59) ͈ˏ_t7ƪǀ5̩4ƬąL ȏ+#0H2˴H. 21H ?+
(60) _Rʚ77ʵ8
(61) ʰ5±ƂL̞. %58Ģˏ'&
(62) ïʟL˜0Ģˏ%H. 2ʢFI0H ±ƂƯ8ŧųęˉʳ̞. νutero-tubal junctionξ5ˏŧ͈̖"I
(63) ʵ5E.08 198 DZC7Ύ͈ˏLˣǐ%H 21HνRacey, 1979ξ Scotophilus heathiν̎ĮπGreater Asiatic yellow batξ18
(64) ͈ˏȂΎ¤8̝¤oeqeoɦƢÆǖęā54H2ɦƢdz%H :ŧųęˉʳ̞5Or^ĢŶÈʑɽ#0H. 2
(65) E. 2ǴF54.0G
(66). 7ʵ5H͈ˏ8oeqeo7ǤͻLĢ0HĦ˻ƹʪļ"I0HνRoy and Krishna, 2010ξ ͈ˏȂΎ¤8\_eɦƢE:\_eqej νGLUTξ7ʑɽÆ
(67) WtlE:WtlqejOCTN2 7ʑ ɽε
(68) ǖęā54H2\_eɦƢE: GLUT ʑɽdz%H "I0G
(69) ͈̖ˏŧ5\_eLÎ˞%H. 2őĴ. 25EG
(70) Ģˏ5äʽ-ˏŧ7ͪċL. Ʌƹď%H2˴FI0HνRoy and Krishna, 2013ξ WʚE:|¯ʚ50 8
(71) ǩƝΎνe.g. Acrochordus javanicusρ̎Į Javan wart snake 18 7 ƝΎξ5K+G͈ ˏL̞. 2ʢFI0HνBirkhead and Møller, 1993ξ. IF7ɱ̚ά18
(72) ±Ƃ. L#04Ɲ1CāƝ5͈̖#+ˏŧLÌ.0ĢˏęLʂA. 21
(73) ͈ˏ8Ģˏ. 7jQ\L̾ˊ%H,14
(74) ĢˏLÔ̶%HƬąCNj.0H2˴FIH ηά18
(75) ˏŧ͈̖ˉνsperm-storage tubules; SSTξ2ķ9IHˉɷ7ȟͤ50͈ ˏ̞KIHνBobr et al., 1964; Brillard, 1993ξ tqDRf7E4ŵʮ18˓ 25 ǶΎ5 1 Ö7ęŧLǖę%H
(76) ęŧĢˏĦ˻4ǶΎ8
(77) ęŧęˉɞǬ5Ũ ň#0HǖęƯ˓ 15 öΎ7@1HνWarren and Scott, 1935ξ ?+
(78) ęŧ͘ęˉ Lͬ͢%Hͬʴ1ęˉ̄ŜF8Ŝ7ęʓöȽ"IH+B
(79) ęʓöȽ"I0 HΎ8
(80) ˏŧ8͘ęˉLͬ͢142ªƻ"IH #+.0
(81) ĢˏLǀć"'H5 8
(82) Ͳ÷4ǶΎ5͘ęˉLͬ͢#Ģˏ7Œ5+3Gʠ. 4. 2ηά508ɶ5̩.
(83) 1H2˴FIH SST 8æʎğƸνDas et al., 2006ξ
(84) ̃F7ǖõνBakst, 2011ξ
(85) ęˉöȽɵ7ɆI˄
(86) ˏŧ52.0ȁŴ4ʀŕFˏŧLΒͅ#
(87) ǖę7jQ\5 īK'0ˏŧLǦõ%HƬąLNj.0GνIto et al., 2011ξ
(88) ͈ˏ8
(89) ηά7Ģˏ5 08ƷΧ7ͬʴ1H ŵʮ508
(90) SST 50͈̖"I+ˏŧ7@Ģˏ1 H2˴FI0H ηά7 SST 8
(91) tqęˉ7ɞǬνvan Drimmelen, 1949ξE:ŧų̃ʳ̞ νutero-vaginal junction; UVJξ5ƃň%HνBobr et al., 1964ξ ʑ̫Ư?C48 sperm-nestsνe.g. van Drimmelen, 1949ξD sperm-host glandsνe.g. Gilbert et al., 1968ξ
(92) sperm-storage glandsνe.g. Burke et al., 1972ξ2Cķ9I0+
(93) ͛Ɲ18
(94) sperm-storage tubules 2 ķ ʲ 5 = > ˠ " I 0 H SST 8 / 0 sperm-host glands D sperm-storage glands 2ķ9I0+͢G
(95) ϧά7ŧų́5Ä+Ėˉɷ́Ƞ7ȟͤL ǐ/ SST 8ƴ˟ȳLȁ%HĖƈ7ʖ1ƪǀ"I0G
(96) SST 7ìƭ8˓ 1020 μm
(97) ˉ8˓ 400600 μm 1
(98) öȐ?+8öȐ7ˉ1HνBobr et al., 1964ξ Ƣ7± Ƃ1Ñç#+ˏŧ͈̖"IHȂΎ8ʵ5E.0ʍ4.0H cllR νMeleagris gallopavoξ18
(99) 815 ͦΎ
(100) tqνGallus gallusξ18 34 ͦΎ
(101) R fνCoturnix japonicaξ1 1112 DZ
(102) O{νAnas platyrhynchosξ18 1015 DZʴ Ƣ2"I0HνBréque et al., 2003ξ ęˉ+G5Ũň%H SST 7ǩCʵ5E.0ʍ 4G
(103) cllR1˓ 20,000 Ö
(104) tq1˓ 15,000 Ö
(105) Rf1˓ 3,500 Ö2 "IνBirkhead and Møller, 1992ξ
(106) 7ǩśʴ
(107) ͈ˏȂΎC2"I0H SST ;Ñç#+ˏŧ8ΨLʛʿÛ5Ǫø#
(108) ȉɷ54.0͈̖"IHνBakst, 1992, Ito et al., 2011ξ SST ʖ˙˺ì58˼˸ɝE:\_^Ωˍ̮Ż"I
(109) )7͛Ý5 8q_rOD`dÈŨň#0G
(110) Ʌʑ5»͂L̞.0H. 2ʪļ"I. 0HνBurke et al., 1972ξ ?+
(111) SST ʖ7ĤÛ18öȽΩˍ
(112) ʛʿÛ18O [ʻ̮͑Ż"I
(113) O[öȽ5EG
(114) \_^ΩˍD˼˸ɝLıAƀ. 5.
(115) ˺ˉ̀ì;öȽ"I0H2˴FI0HνSchuppin et al., 1984; Bakst and Bauchan, 2015ξ 7E5
(116) åũΫƴΉDΞŧΫƴΉLʃ+̮Ż˝ȏ8 50 Ɲ¼āFőĴ "I0H žˏƯ7ˏŧ8 SST 5Ñç#
(117) ȂΎÈɕº͛1Ģˏ˻Lˣǐ#+??͈̖"I
(118) ǖęā5 SST FǦõ"IHνSasanami et al., 2013ξ %4K-
(119) ηά5H͈ˏ8
(120) SST ;7Ñç
(121) SST ì17͈̖
(122) SST F7Ǧõ2 3 /7ȭΗFǀGʽ.0H 2˴H. 21H SST ;7ˏŧÑçLþƲ%Hʜǘʔ4̩ł8ǴF54.0. 4
(123) Rf508
(124) ˥ǖȼĤ́7öȽɵ5ı?IHej\d F2α SST 7ĤLǎƧ%H. 25EGˏŧ7ÑçLÓ#0H. 2őĴ"I0H. (Sasanami et al., 2015) ?+
(125) SST F7ˏŧǦõ5/08
(126) Ito et al.ν2011ξ5E. 0Wtf7ʿőĴ"I0H ηά18˓ 25 ǶΎĵȂ1ǖę͑. G
(127) )I. 5Â.0̝¤^eoɦƢdz%H SST ʖ˙˺58 membrane progesterone receptor ανmPRαξʑɽ#0G
(128) ^eo7ÿɤ5EG SST Ġ˪%H 5EGɵɿʔ5͈̖ˏŧLǖõ%H. 2. 2őĴ"I0HνIto et al., 2011ξ ?+
(129) . ^eo7dž 5EG
(130) UVJ 7ʖ˙˺F[l[Ǧõ"IH. 2C̮Ż"I. 0GνIto et al., 2011ξ
(131) [l[8ŧų5Ũň%Hęȯ7º+ę7Ł͕LÓ%ɡ ɜȻ7E4ƬąLȏ+#
(132) ę2ŧų7ìŗ27ΎLˏŧͬ͢%H7L̥ĉ#0 H2˴FIH ^eo SST F7ˏŧǦõłŧ2#0à. 28
(133). ǖę2ˏŧǦõ
(134) E:ˏŧ7ͬ͢L̥ĉ%H[l[öȽ7jQ\Lĭ̾"'H 2ɪ1ɿ5Ͳ.0G
(135) ε˄ċɵ7ʁȬǁʋ2#08Ρƚ5̊ĶɎ ##4 F
(136) ̰Ǵ"I04ɪCȫ"I0H Í9
(137) ^eo5ğƸ%H SST 8 èÈ71G
(138) ͈ˏȂΎL͢$0Ɓ#&/ˏŧǦõ"IH. 2̮Ż"I0H. νIto et al., 2011ξ
(139) )7Wtf8ǴF24.04 Ĉ0
(140) 4( SST 5͈ ̖"I+ˏŧ8ȂΎĢˏ˻Lˣǐ1H72Ľ58
(141) Ȅ,ˇõ04. 6.
(142) ˏŧ8jz[īǀL̞K45CKF&
(143) SST ì137E5#0ʁŨ#
(144) Ģ ˏ˻Lˣǐ#0H8
(145) Ɲ5K+Gʤʹ˵7̊Ķ7ż͆1Gˢ0H ˏŧ͈ ̖ˉ5ʬ˜džž̮Ż"IνFreedman et al., 2001ξ
(146) ?+
(147) O|dνFoye-Jackson et al., 2011ξ
(148) O[OνZaniboni and Bakst, 2004ξ˄ƃň#0G
(149) ͈ˏ;7ƫΤL˴ Ż%HőĴ8H
(150) ͈ˏ27ʜǘʔ4Ώͥƹ8̶Ǵ"I047ɽɷ1H ? +
(151) tq50
(152) ˏŧÑç#+ SST 18ƪ͍͕ǝǀłŧβTGFβ7ʑɽ dz%H. 2őĴ"I0HνDas et al., 2006ξ ęˉ8
(153) ̰Ăũʔ58ÈŚ5ʝƩ%. H+B
(154) ęˉì5Ñç#+˙̑437ʍɵLǖΕ%H+B7æʎʑͮ#0G
(155) ęˉ ì5Ñç#+ˏŧCʍɵ2#0æʎ˙˺5ǥǟ"I0#? TGFβ18 T ˙˺D B ˙ ˺7ŖȬLDžþ%H. 2ηά1ʢFI0GνQuere and Thorbeck, 1990ξ
(156) SST 5͈. ̖"I+ˏŧ5ż%HæʎğƸLDžHàL%H2˴FI0H Re7Ģˏ5 0C
(157) Kawano et al.ν2014ξ 5E.0
(158) ĭȠ7¸˛@ʑ̫"I+ %4K-
(159) ̃ ì5žˏ"I+ˏŧ8ŧųì7̥È5ǥǟ"IH
(160) ˏɠ5ı?IH seminal vesicle protein 2νSVS2ξ5EGˏŧ̣LÔͅ"I
(161) 7̥ÈFǥǟLŁͶ%H¸˛@̫ õ"I0H űΘ5
(162) SVS2 7wm[ORqRe8Ģˏ7Œ1Hęˉ̄Ŝ5ý ͮõȋ4+BΙƹŢ24H
(163) ΫƴĢˏ18ʂ·ƱFIH ##
(164) IF7Λ ƹʁȬͭ7ǥǟFˏŧLÔͅ%HWtf8
(165) ʷȝʔ5ˏŧ7Ģˏ˻LȂΎˣǐ %H¸˛@28˴Ν SST 5͈̖"I0Hˏŧ8ͪċLÚȦ#0H2"I0GνBakst, 1987ξ
(166) ˏŧ 8ͪċLÚȦ#0»͂LDžþ%H. 25EG
(167) Ʌƹ;˖5EH˙˺̂7ßŴLɓƁ"'. 0ȂΎ7͈̖5˶0H71842˴FI0HνBréque et al., 2003ξ ɶ 5̊ĶɎ78
(168) ηά7ˏŧ8
(169) ͘ęˉ7¤1ΠȦɷƾFͪċɷƾ;7÷GǿL 2 ŁνžˏƯE: SST F7ǦõƯξ̞. 21H. IF7ͪċ7ÚȦŤL. þƲ%HöŧeQml7ÊċȣƠ8
(170) ɶ5ÈìĢˏL̞ċɵ18=2M3ǴF54. 7.
(171) .04 OWxQνCynops pyrrhogasterξ50ˏŧ7ͪċL̺͑%H SMIS νsperm motility-initiating substanceξĭů"I0GνWatanabe et al., 2010ξ
(172). I. ÈìĢˏċɵ50ùB0őĴ"I+ˏŧͪċŤłŧ1H OWxQ18
(173) VeˏĎLǦõ#
(174) eˏĎLġG͚@͘ęˉ7ĢˏĿ5͈̖%H ǖę"I+ę ŧ5EGĢˏĿFlj#õ"I+ˏŧ8
(175) ęŧ̣7hƈ5ǘ̱%H2ͪċLŤ %H hƈ̣7Ωˍì58 SMIS ƃň#0G
(176) ˏŧhƈ5ýͮ%H2 äÈğƸ̺͑"I
(177) äÈì7ͽ˖5EGΩˍFǦõ"I+ SMIS ˏŧ7ͪċL Ť"'H. 2ʪļ"I0HνWatanabe et al., 2010ξ OWxQ7E5
(178) ˏ. ŧͪċLŤ%Hā5äÈğƸ̺ſ"IHÍ8¹5άL̫4C71
(179) ̋ʔ4È ìĢˏċɵ18
(180) ˏŧ8ͪċ˻LƱ+7-ęˉLͱ#
(181) ęŧ5ǘ͛#äÈğƸL͑ % . I?17ʤʹ1
(182) UVJ 7NJõɵLɑĈ#+ŎΰɌ1ˏŧLQYq%H2
(183). ˏŧ7ͪċDžþ"I
(184) ˏŧ7ͪċˡˢǶΎƣ%H νMatsuzaki et al., 2014ξ Ʌƹɵ͍ı?IH. 2ǴF24.0H. I8
(185) UVJ 7NJõɵ5ˏŧ7ͪċLÆ"'HȄʢ7ʁɿ. 2Lʪļ%HC71H ). 1Ȇʤʹ18
(186) SST 8ˏŧ7ͪċ. LɅƹď%Hɵ͍ı?I0H2½̼Lʽ0
(187) 7ɵ͍Lĭů#
(188) )7Êʃȣ ƠL̰Ǵ%H. 2Lʚʔ2#+. 8.
(189) ˁ¬ʾ ˏŧ͈̖ˉʅȋˏŧͪċDžþłŧ7ĭůE:ˏŧ͈̖ˉ5H §;ʂʁȣȟ. ˁˊ ˧̿ ηάLıAś7ÈìĢˏċɵ18
(190) ±Ƃ2Ģˏ7jQ\8Ʒ&#C̉'&
(191) ˏŧ2ęLͲ÷4ǶȂ2Œǂ5êŨ"'H+B7̾ˊȣȟŨň%H 7ǶΎ̾Ǫ7 ƬąLNj.0H7͈ˏ1G
(192) ͈ˏL̞ċɵ18žˏƯ7ˏŧ8ęˉì1ůȂ Ύ͈̖"I
(193) ǖę?17Ύ
(194) Èɕº͛1CĢˏ˻LÔ.+??ˣǐ"IH ͈ˏ7ȂΎ 8ʵ5E.0Ƞ1
(195) Í9ŵʮ508
(196) Rf1 11 DZνSittman and Abplanalp, 1965ξ
(197) tq1 35 DZνNalbandov and Card, 1943ξ
(198) cllR1 117 DZνChristensen and Bagley, 1989ξ2őĴ"I0H ηά5H͈ˏ7Œ8
(199) ęˉ7ŧų̃ʳ̞ νutero-vaginal junction; UVJξE:ɞǬ5ƃň%Hˏŧ͈̖ˉνsperm-storage tubules; SSTξ1HνBobr et al., 1964ξ žˏƯ7ˏŧ8 SST 5Ñç#+7-ͪċLÚȦ#
(200) Ģˏ˻LÔ.+??Èɕº͛1ȂΎˣǐ"IHνBakst, 1987ξ ɶ˃%<8
(201) ˏŧ 8͘ęˉ7¤1ΠȦɷƾFͪċɷƾ;7÷GǿL 2 ŁνžˏƯE: SST F7 ǦõƯξ̞. 21H Rf18
(202) SST FǦõ"I+ˏŧ8 UVJ 7ˎ̂ʖ˙. ˺FǦõ"IH heat shock protein 70 5EGͪċLíɅƹď%HĦ˻ƹʪļ"I0 HνHiyama et al., 2013ξ
(203) )7¹7ͬʴ50ˏŧ7ͪċLþƲ%H̩ł8Ǵ 1H . I?15
(204) SST 5Hˏŧ7ͪċÚȦ7ʁɿũʔƬą˴Ż"I0H. νFroman, 2003; Froman and Feltmann, 2005; Froman and Feltmann, 2010ξ
(205) SST ͈̖ˏŧ5 HͪċÚȦ7öŧWtf5/08ǴF54.04 Ȇʤʹ18
(206) SST 8ˏŧ7ͪċLɅƹď%Hɵ͍ı?I0H2½̼Lʽ0
(207) 7ɵ͍Lĭů#
(208) )7ʂʁȣƠL̰Ǵ%H. 2Lʚʔ2#+. 9.
(209) ˁ¬ˊ ȇǭĝ:ǰȾ (1). űγċɵ. qYνǂȺƗξF͎ç#+ 7 ͦμ7Rf (C. japonica)L
(210) ³ƔɭǴ5EG 14 ǶΎǴȂ10 ǶΎǻȂ7åĵȂ
(211) Ųɕ 25C 7Ȋ¾1ί˹#+ α8
(212) qYF͎ ç#+ǀιʃίǭLǮ˞α1
(213) ȵ8ȵͭȵLǮ˞ȵ1 + ʂęǶĀLÖÈ û5̴·%H. (2). 25EGĪÖÈ7ǖęªůǶĀLǙů#
(214) űγ5ʃ+. ̸̘. ;˩̡ʁɿήŔȵνPBS: 137mM NaCl, 2.68 mM KCl, 8.10 mM Na2HPO412H2O, 1.47 mM KH2PO4, pH 7.4, 282 mOsm/Lξ
(215) 8
(216) è0ĸå˔̘ƔȜνŜΑƗξF͎ç# +̸̘Lʃ0̦̾#+ x[eƜ̢ŔάɛɌνHBSS: 137 mM NaCl, 5.37 mM KCl, 1.26 mM CaCl2, 0.81 mM MgSO4•7H2O, 0.34 mM Na2HPO4•2H2O, 0.44 mM KH2PO4, 4.2 mM NaHCO3, 5.56 mM glucose, 0.01% phenol red, pH 7.4, 306 mOsm/Lξ8
(217) Q~o[wd fνɗđξF͎ç#+ HBSS 10 ×ɌνWcR
(218) \vcR
(219) ɩ;ȵ˖sq R
(220) ~Swmrıξ5
(221) ā͝7ɦƢ54HE Milli-Q ȵ1ƘͿ#
(222) CaCl2, MgSO4•7H2O, NaHCO3 E:~SwmrLɑĈ#0Ìʃ#+. (3). UVJ NJõɵ7̦̾. ͥʂeRfLǮΨƇȮ5EGūȞȨ"'+7-
(223) ͘ęˉLǞõ#+ ͘ęˉLŧ ųF̃50÷#
(224) ŧų2̃27ŕʉ5Åˮ%H UVJ LǗġ#+ UVJ LʁɿήŔȵν0.9% NaClξ¤5ġG
(225) SST Lı?4ÅLűÈΫƴΉ1ΕĜ#+ q\#+ UVJ LTmr~l5çI
(226) 500 mg ˛ˬ/ml 54HEȶ ð PBS LĈ+Ư
(227) ̰Ăya1˙÷#
(228) ͲŰ om[eYa1ǣnj#4F ȶ1 2 ǶΎNJõ#+ 20,400 × g
(229) 4 ˚C 1 10 öΎͰƶöΜ#
(230) ɒL UVJ NJõɵ2. 10.
(231) #+. (4). ˏŧ7ǗġE:ͪċOmgQ. žõˏŧ8
(232) Kuroki and Mori ν1997ξ7ǰȾ5ư.0Ǘġ#+ Ǘġ#+ˏŧ8 HBSS 5ƿɥ#+7-q̝ɾ̳ˈʘLʃ0ˏŧɦƢLˈõ#
(233) ɦƢ 2.0 × 107 sperm/ml 54HE HBSS 1ƘͿ#0OmgQ5ʃ+ ˏŧ7ͪċƹL̷Ð%H+B5ˏŧͪċOmgQL̞.+ UVJ NJõɵL 10%Ŷ ıA HBSS 7rmLelm[c5Êǀ#
(234) Ɇċz~P1Žç#+ Êǀ#+rm8
(235) OmgQ5ʃHā5 39C 1ªɕ#+ ͝7ǰȾ1Ǘġ#+ ˏŧƿɥɌLrm7 20%ŶĈνȀ˚ˏŧɦƢ 4.0 × 106 sperm/mlξ
(236) mq q1 39 ˚C 5Ô-4FűÈΫƴΉνM165FCρQWQ[ceof
(237) ǯŷ đξ1̮Ż#+ ˏŧ7ͪċƹ8
(238) 4 νɤ#āͧͪċξ
(239) 3 νɅʑ4āͧͪċξ
(240) 2 νD DƦāͧͪċξ
(241) 1 νƴƦ4āͧͪċξ
(242) 0 νͪċÚȦξ7®ȭΗ1̷Ð#+νWheeler and Andrews, 1943ξ _q˱2#0 10% PBS LıA HBSS Lʃ
(243) ĭȠ5ˏŧ 7ͪċƹL̷Ð#+. (5). ^ɧͬ[q\~P. ^ɧͬ[q\~P5Î%H UVJ NJõɵ8
(244) Z;OtRym~N ν20 mM Z;OtR
(245) pH 7.4ξLʃ͝7ǰȾ1̦̾#+ UVJ NJõɵ8 Superdex 200pg νGE e]O
(246) DZƗξWν⌀ 1.6 × 75 cmξLʃ0öΜ#+ ˾ȴ#+Z;OtRym~N240 mlν2 WŶʷöξ1WLƜ̢ď#+ 7-
(247) UVJ NJõɵ 1 ml LW5OQ#
(248) "F5Z;OtRym~N140 ml LɆͣ 1.0 ml/min 1ɛõ"'+ ɛõɌL 7.5 ml &/öġ#
(249) Īöʈ5/0ˏŧ ͪċOmgQ5EHe[t\L̞.+ %4K-
(250) ƱFI+öʈLò˝¨ɰ#+. 11.
(251) 7- Milli-Q ȵ5íɛ̰%H. 21ɦ˪#
(252) HBSS 5 1% ŶɑĈ#0ͪċOmgQ5. EHe[t\L̞
(253) 24 ǶΎƯ7ͪċƹε.+öʈLΚBī#+ Superdex 200pg 7 QrŶ (V0) E:öŧLˈõ%H+B7ȡɚǽ˨8
(254) Ferritin (440 kDa)
(255) Aldolase (158 kDa)
(256) Conalbumin (75 kDa)E: Ovalbumin (43 kDa)7ɛõÈ ʷ5EGÊǀ#+. (6). εͣɌÈ[q\~PνHPLCξ. UVJ NJõɵ7 Superdex 200pg ÆöŧöʈνɛõÈʷ 100110mlξLò˝¨ɰ#+7 - Milli-Q ȵ5íɛ̰%H. 21ɦ˪#
(257) C-22 WνNavi C22-5ρ4.0 × 150 mm, ĸå. ˔̘ξLʃ+ HPLCνƎɄ̦Êǂ
(258) ²ͺƗξ5EGöΜ#+ ʳċʝ A L 0.1% q ~Vͼ;νTFAξLıA 80%Ogqtq
(259) ʳċʝ B L 0.1% TFA LıA H2O 2#
(260) A : B = 0 : 100 1 20 öΎɛõ#+Ư
(261) A : B = 0 : 100 A : B = 80 : 20ν20 öΎξ 7tO\dTq1ɛõ#
(262) ȀƯ5 A : B = 100 : 0 1 5 öΎɛõ#+ 220 nm 5 HIJåLțõ#
(263) _k1̴·%H22C5
(264) }[̮Ż"I+ɛɌLöġ# + ƱFI+öʈ8ò˝¨ɰ%H. 25EG TFA LΕĜ#
(265) Milli-Q ȵ5íɛ̰#+7. -
(266) HBSS 5 1% ŶɑĈ#
(267) ͪċOmgQ5EHe[t\L̞.+. (7). öġ[q\~PνPLCξE:̗ƈ[q\~PνTLCξ. PLC 8
(268) PLC Xeq cW^ 60 F254νŅůƈĚ 1 mmρ[
(269) ʚλđξ Lʃ0̞.+ Y}Lʃ0q5aLemq#
(270) ͼ;Tl : jw : H2O = 6 : 4: 1 7ɛť1ƅ#+7-
(271) qLpc]j¤1¨ɰ" '+ aLemq#+ÅˮF 1cm !27đʈ5ö
(272) qFcW ^LǚġGXe̸γˉ5öġ#+ cW^5ƅɛťLĈ
(273) ɤ#ǣnj# 4F 30 öΎNJõ#+Ư
(274) ɛťLɧͬ#
(275) ʼ˖XeLijºH. 12. 25EGɛťLʊ.
(276) Ĝ#+ ˝ǹ?+8VQɷ7ɌÈL 10 μl 7 Milli-Q 5ɛ̰#+7-
(277) HBSS 5 1%Ŷ ɑĈ#
(278) ͪċOmgQ5EHe[t\L̞.+ PLC 5EGƱFI+öʈ5ż#
(279) TLC öȍL̞.+ Y}Lʃ0 TLC O cq cW^ 60 F254ν[
(280) ʚλđξ5aLemq#
(281) ͼ;Tl : jw : H2O = 6 : 4: 1 7ɛť1ƅ#+ ƅƯ
(282) qLݨ#
(283) p;ʑ̸̘̍ν5% 12MoO3ςH3PO4, 100%TjwɛɌξ5qLɊɟ#+7-
(284) mqq1Ĉɯ%H. (8). 25EGʑ̍"'emqL̮Ż#+. NMR. 1H-NMR
(285) 13C-NMR
(286) E:¬ȥã NMR e[qνHMQC E: HMBCξ8
(287) JEOL GSX-500 Ȗʩȴêθ̤ˮν500 MHzρDZȆΞŧ
(288) ǵƎƗξLʃ
(289) ȵνD2Oξ5ɛ̰ #+̸ǭLɖů#+ ƱFI+e[qpjL̰ȍ#+7-
(290) L-§;νc\O rmlξ7e[qpj2ɭī#0̉%H. (9). 2Lʨ̹#
(291) ďīɵLĭů#+. SST 7§;ů. ͥʂeLǮΨƇȮ5EGūȞȨ"'
(292) ͘ęˉLǞõ# UVJ LǗġ#+ Ǘġ#+ UVJ ˎ̂L OCT _zRrνa[~NQom[dz
(293) ¤Şđξ5ĎŌ#
(294) 80˚C 1ò˝#+7-
(295) [VejmqLʃ0Ě" 30 μm 7ò˝÷ɳLÊǀ#+ bQ[kQg[cνPALM CombiSystemρWnNQe
(296) ǯŷđξLʃ 0 UVJ ò˝÷ɳF SST ?+8 UVJ ʖ˙˺L÷Gõ%22C5
(297) bLɭž #+ʷ2ò˝÷ɳ7Ě"F
(298) ÷Gõ#+˙˺œ7ÈʷLˈõ#+ ŁĠ#+˙˺œ 5 100 μl 7 Milli-Q ȵLɑĈ#0
(299) qdsQbLʃ0dsQf# +7-
(300) dsQf#+ɛɌL 20,400 × g
(301) 4˚C 1 10 öΎͰƶöΜ#
(302) Tyj Lʃ0ɒL¨Ņ#+ Milli-Q ȵLĈ0íɛ̰#+Ư
(303) L-Lactate Assay KitνCayman
(304). 13.
(305) cXƒξLʃ
(306) ̼ǴǾ5ư.0§;ůL̞.+ %4K-
(307) ͽ˖ɛɌ
(308) ̥ͽ˖ ɛɌ
(309) ̛åŏ͍LıAOmgQym~N5aɛɌLɑĈ#
(310) 20 öΎŲɕ1ğ Ƹ"'+ rrY}1ğƸɌ 4 μl L̳G2G
(311) ̛åΫƴΉνBX51ξE : Image JνOWņʽ̠ʁʤʹǂ
(312) rƒξLʃ0rrY} ì7ğƸɌʑ%H̛åLů#+ bQ[kQg[c5EG÷Gõ #+˙˺œ7ÈʷF
(313) SST ?+8 UVJ ʖ˙˺5H L-§;ɦƢLˈõ#+. (10) SST 7ĖΜ SST 7ĖΜ8
(314) _^shôɿLʃ+ǰȾνKing et al., 1999ξ5EG̞.+ ͥ ʂeLǮΨƇȮ5EGūȞȨ"'
(315) ͘ęˉLǞõ# UVJ LǗġ#+ űÈΫƴΉ νM165FCξ1 SST Ũň%HÅLq\#+7-
(316) SST LıA UVJ ˎ̂L l5Ǘġ#+ _^shɛɌν10 mg/ml _^shνĸå˔̘ƔȜξL ıA HBSSξL 1 mg ˛ˬ/μl ɑĈ#̰Ăya1˙÷#+7-
(317) _^shɛɌL" F5 500 μl Ĉ
(318) ǣnj#4F 37C 1 20 öΎQYq#+ _^shô ɿƯ
(319) ȶð#+ğƸÚȦɌν10 mg/ml ɴ̝ɒOLıA Ca2+/Mg2+ı HBSSξ L 5 ml ɑĈ#0ͽ˖ğƸLÚȦ"'+ ğƸɵLc5ʳ#
(320) Q[}mq LʃűÈΫƴΉ1ĖΜ"I+ SST LŁĠ%H22C5
(321) SST Lı?4 UVJ ʖ ˙˺CĭȠ5ŁĠ#+. (11) SST 7§;ʂʁ˻ɖů SST 7§;ʂʁ˜͔L̾<H+B
(322) ̰ː˒LΓŴ#+Œī5H SST 7§;ʂʁ ˻Lɖů#+ %4K-
(323) ĖΜ#+ SST L 1 ml 7 HBSSν\_eı 5 .56 mMξ
(324) 2-DOG Ɍν1 mM 2-pVYc\_eLıA\_eı HBSSξ?+8 2-DOGal Ɍν1 mM 2-pVYcX[qeLıA\_eı HBSSξ¤50 39C 1Ŏ. 14.
(325) ΰ#+Ư
(326) Πˮ#0 SST Lȹɣ"'
(327) ŎΰɒLŁĠ#+ ŎΰɒL 20,400 × g
(328) 4˚C 1 10 öΎͰƶöΜ#
(329) ɒ5ı?IH L-§;ɦƢL
(330) L-Lactate Assay Kit νCayman
(331) cXƒξLʃ0͝7ǰȾ1ů#+. (12) RT-PCR RNAisoνjWyQV
(332) ̏ɄƗξLʃ
(333) _^shôɿ5EGĖΜ#+ SST Fè RNA LNJõ#+ ɐç#+ DNA L DNase IνjWyQVξ5EGö̰#+Ư
(334) ReverTra AseνȌɂ˗
(335) ŜΑƗξLʃ0͕͠îğƸL̞.+ cDNA LΆŋ2#
(336) MCT1 νgeπ5 - GAACCCTGCCTTAACCATGA - 3
(337) Olgeπ5 - GGCACA CCCCATTGTAAATC- 3 ξ
(338) MCT2νgeπ 5 - TGTGGCTGGGTCACTTATGA - 3
(339) Olgeπ 5 - CTGGCCACAATCACAATCAC- 3 ξ
(340) MCT3ν5 - CTGGGTA TGGCGTTGAACTT - 3
(341) Olgeπ5 - TAAGTAGGACCAGCCCGATG- 3 ξ? +8 MCT4νgeπ5 - AGCACCAAGCCAATAGCACT - 3
(342) Olgeπ 5 GGGTCTGGCACTCAACTTTC- 3 ξ7ɶʍʔQLʃ0 PCR L̞.+ ì ȡɚ2#0
(343) Rf S17 ijz[͵Áŧ7Qνgeπ 5 - GAC GAAGACGGTGAAGAAGG-3
(344) Olgeπ5 -CTTGGTGTCTGGGTCCACTT-3 ξ Lʃ0ĭȠ5 PCR L̞.+ vXoP_q˱2#0
(345) ͕͠îͽ˖L Milli-Q ȵ5ˮǝ#0͕͠îğƸL̞.+Ư
(346) MCT1
(347) MCT2
(348) MCT3 ?+8 MCT4 7ɶʍʔ QLʃ0ĭȠ5 PCR L̞.+˱L̵+ PCR ʂɵL
(349) ̇ďTldRLıA 1%OXe^¤1ΞȴɁċ#
(350) Ħ̬ď"I+yrL LAS 500νGE e]Oξ 5EG̴·#+Ư
(351) Image J Lʃ0yr7ůL̞.+. (13) in situ xQkQhc. 15.
(352) in situ xQkQhc5ʃH UVJ ò˝÷ɳ8
(353) Yoshimura et al.ν2000ξ7 ǰȾ5EGÊǀ#+ MCT4 7Olgeν5 - CGCAAAGGAACACTGG GCTCCCAGCAGCAGACAACCCGTTGGCCA -3 , 5 - ATTGCTTCTGCCAGGAGCA CCAAGCCGATAGCACTG -3 , 5 - AAACGCCGTGCGGGTTTTGTGCCGTGCTCAC ACACA -3 , 5 - TGTGACCAGCTGAGGGCAGTGCCTGCGATGTGCCTTTC -3 ξ8
(354) jspVYc u[Vldqe~Shνa~PmcaQ ToP~Pm[ξ5EG[33P] dATPνNEN Q~aQTek[n
(355) al gmnƒξȡ̈́#+C7LÌʃ#+ 42C 1ǷxQkQhc#
(356) Ųɕ1 30 öΎ7ɃɇL 2 Ł̞.+7-
(357) 55˚C 1 40 öΎğƸ"'+ eQr\eLɃɇ# +7-
(358) X ˨~PνźŘ~P
(359) ɗđξ5ƽå"'
(360) űÈΫƴΉνM165FCξ1 ̮Ż#+. (14) RNA-Seq ʃ cDNA Q7ȟˌ RNA-Seq ̰ȍ5ʃHeqrɶʍʔQ7ȟˌ8
(361) SureSelect strand-specific RNA library prep kitνOdqo[wd
(362) W~UtOƒξL ʃ
(363) ̼ǴǾ5ư.0̞.+ è RNAν400 ngξFνAξͻøLıA mRNA L ŁĠE:Ǯɳď#
(364) O[lwQc Dνc\Orml
(365) ʚλđξ7Ũňď1 Ǯɳď#+ mRNA L͕͠î#+7-¬ȆΈ cDNA Līǀ#+ cDNA 7ȅʿÕƳE :OptºĈL̞.+7-
(366) SureSelect Oligo Adaptor LQ^c# cDNA 5O kjͻøLºĈ#+ Qpm[eQLıA 3 /7QLʃ0O kjºĈ cDNA L PCR 5EGŖƛ#
(367) RNA-Seq ̰ȍ5Ìʃ%H cDNA Q 2#+ MiSeqνQs
(368) ɗđξLʃ0 SST E: UVJ ˎ̂ʖʅȋ cDNA Q 7c]c\ν76 bp
(369) OTrξL̞.+. 16.
(370) (15) RNA-Seq
(371) de novo OgE:pP~Nc̰ȍ cutadaptνMartin, 2011ξLʃ
(372) èrͻøFOkjͻø
(373) Æ[VoPνQV < 30ξ7ȅʿE: 76 ʌʚ7ȅʿLΕĜ#
(374) 50 bp ¼7ʣrLʥȚ%H. 25. EG
(375) rL[Om#+ ͛Ɲ
(376) ȥ¢»c]aLʃ0Rf7r ~q^w̰̽"I0HνGenBank ʒ·ʌĨπDRA000595ξνKawahara-Miki et al., 2013ξ ). 1
(377) ùB5Rfr~q^wL~Ne2#0OgL̞
(378). Tophat E: Cfflinks Lʃ0qe[qpj7pP~Nc̰ȍ L̞.+νTrapnell et al., 2012ξ ##4F
(379) ~Ne^w2#0ʃ+Rf r~q^w18ʑɽƖLțõ%H+B7rēö5m}\14. + Rf508
(380) ȥ¢»c]a7DŽ̟2Ȳ͗#0
(381) Trinity 1ʁǀ#+ε[ VoP7qe[qFOg#+r RNA-Seq ̰ȍ5Ͳ#0 H. 2ǴF54.0HνFinseth and Harrison, 2014ξ #+.0
(382) Trinity 1q. e[q7 de novo OgL̞
(383) Og#+ͻøL~Ne 2#0ʑɽƖ̰ȍL̞.+ %4K-
(384) Trinityνversion: trinityrnaseq-r20140717ξ νGrabherr et al., 2011ξLʃ0[Om#+r7 de novo OgL̞
(385) megablast 5EG SILVA rRNA pjeνQuast et al., 2103ξ2ɭī#
(386) rRNA ʅȋ7ͻøLΕ Ś#+ Trinity 7\QLʃ0 rRNA ͻøLΕĜ#+rL
(387) BowtieνLangmead et al., 2009ξLʃ0m}\#+7-
(388) RSEMνLi and Dewey, 2011ξLʃ0͕î ɵ7ŨňȲLǙů#+ SST 2 UVJ ˎ̂ʖ7͵Áŧʑɽ7Ɩ8 edgeR zm]d νRobinson et al., 2010ξLʃ0ȸů#+ ?+
(389) ̰ː˒E:Æ;˖5Ώͥ%H͵Á ŧgmq7 SST 2 UVJ ˎ̂ʖ5HʑɽƖLʨ̹%H+B
(390) PAGE ̰ȍνparametric analysis of gene set enrichmentξνKim and Volsky, 2005ξL̞.+ qe[q 7 GOνgene ontologyξOwoc8
(391) InterproScanνhttp://www.geneontology.org/ external2go/interpro2goξLʃ0̞.+. 17.
(392) (16) ˛ˬÆ;˖7țõ ͥʂRf5 180 mg/kg BW 7 HypoxiprobeTM-1νďīɵĮπpimonidazole HCl
(393) sl ~NcOQjscs
(394) algmnƒξLΠ˽ɀž#
(395) 2 ǶΎƯ5ǮΨƇȮ#0 UVJ LǗġ#+ _q˱2#0
(396) ʁɿήŔȵLĭΠ ˽ɀž#+˱L̵+ Ǘġ#+ UVJ L OCT _zRr5ĎŌ#
(397) 80˚C 1ò˝" '+Ư
(398) [VejmqLʃ0Ě" 10 μm 7ò˝÷ɳLÊǀ#+ ò˝÷ɳLȶðO gq1Ņů#+7-
(399) ɑº7̼ǴǾ5ư
(400) LJ pimonidazole LJÈLʃ+æʎ˛ˬď ũȒ̍L̞.+ 1 μg/ml 7 DAPI 1ȖȒ̍#+Ư
(401) ̛åΫƴΉνBX51
(402) Vze
(403) ǯŷđξLʃ0̮Ż#+. (17) q_rOɅƹɖů SST 7q_rOɅƹɖů8
(404) JC-1νďīɵĮπ5,5 ,6,6 -tetrachloro-1,1 ,3,3 -tetraethylbenzimidazolylcarbocyanine iodide
(405) a~PmcaQToP~Pm[
(406) algmnƒξLʃ0̞.+ %4K-
(407) ͥʂeLǮΨƇȮ#+7- UVJ LǗġ#0l5ġG
(408) 50 μl 7 HBSS LĈ̰ĂyaLʃ0˙÷#+ JC-1 LıA HBSS L 500 μl ĈνȀ˚ɦƢ 125 nMξ
(409) 37˚C 1 20 öΎQYq#0̍ ˖LpP\#+7-
(410) 800 × g
(411) Ųɕ1 2 öΎͰƶöΜ#0 UVJ ˛ˬɳLȹɣ" ' 1 ml HBSS 1Ƀɇ#+ _q˱2#0
(412) ɃɇǶ5q_rOì̂7˾ê Ƭă1H carbonyl cyanide m-chlorophenylhydrazoneνCCCPρc\OrmlξL 10 μM ɑĈ#+˱L̵+ 800 × g
(413) Ųɕ1 2 öΎͰƶöΜ#+7-
(414) ȹɣ#+ UVJ ˛ˬɳLeQr\e5Rq#
(415) ̛åΫƴΉνBX51ξLʃ0̮Ż#+ ʈá LΫƴΉʃpdjWνDP70
(416) Vzeξ5EG̴·#+Ư
(417) Image J Lʃ0. 18.
(418) ̛åƨƢLůď#
(419) 590nm E: 530nm 5H̛åƨƢ7ȲνFI ȲπF590/F530ξL ˈõ#+ FI Ȳ7ˈõ8
(420) SST E: UVJ ʖ˙˺5/0̞.+. (18) ˠ̳̰ȍ űγ˱Ύ7ȁƼƖ8
(421) öǨöȍȾE: Tukey 7śțůȾ5EGúů#+ ˏŧ7 ͪċOmgQL̞.+űγ5/08
(422) Mann-Whitney 7 U țůȾ5EGˠ̳ʔ4ȁƼ ƖLúů#+. 19.
(423) ˁˊ ˝ȏ (1). ^ɧͬ[q\~P5EH UVJ NJõɵ7öΜ. UVJ NJõɵFͪċDžþłŧLöΜ%H+B
(424) ?&^ɧͬ[q\~P5E HöʈL̞.+ UVJ NJõɵL Superdex 200pg WLʃ+^ɧͬ[q\~ P5Î%H2
(425) ɛõÈʷ 45 mlν}[ 1ρöŧ 1,000 kDaξ
(426) 70 mlν}[ 2 E : 3ρ50 kDaξ
(427) 100-120 mlν}[ 4 E: 5ρ10 kDa ¼ξº͛5¥̩4}[ț õ"I+νFig. 2-1ξ ƱFI+Īöʈ5/0ˏŧͪċOmgQL̞
(428) 24 ǶΎƯ7ˏ ŧ7ͪċƹL̮Ż#+2. J
(429) }[ 4 LıAɛõÈʷ 100-115 ml 7öʈLɑĈ#+. ˏŧ8
(430) ͪċDžþ"I+ (pj8ʪ#04) 7öʈL
(431) HPLC 5EG"F5 öΜ#+. (2). HPLC 5EH UVJ NJõɵ7öΜ. UVJ NJõɵ7 Superdex 200pg }[ 4 L
(432) C-22 WLʃ+ HPLC 5EGöΜ# + ɛõÈʷ 0.7 ml F 2.0 ml 5˖͢G7}[țõ"I++B
(433) ń5ʪ#+ 11 ö ʈLöġ#+νFig. 2-2, Aξ. IF7öʈLò˝¨ɰ-í̜̰5EGɦ˪#+Ư
(434) ͪċ. OmgQ5Î%H2
(435) öʈ 2 5ˏŧ7ͪċDžþɅƹ̹BFI+νFig. 2-2, Bξ C-22 W5ȩ3Ôǐ"I4˖͢Göʈ58ȝƹεɵ͍ı?IH2ªƻ"I++ B
(436) Φʝ˒7 PLC 5EHöΜL̸@+. (3). PLC 5EH UVJ NJõɵ7öΜ. C-22 7ͪċDžþɅƹöʈL PLC 5Î#
(437) cWLöġ#0NJõ#+Ư
(438) TLC 5E. 0öΜLʨ̹#+ TLC öȍ18p;5EHʑ̍@FI
(439) öʈ 7 E : 8 50ʡĘ1ʪ#+Ė7emqțõ"I+νFig. 2-3ξ Īöʈ5/0ˏ ŧ7ͪċOmgQL̞.+2. J
(440) öʈ 7 E: 8 5ͪċDžþɅƹ̹BFI+νFig.. 20.
(441) 2-3ξ öʈ 7 E: 8 Lī#0ò˝¨ɰ#+2. J
(442) VQɷ7ɌÈ24.+ 7. öʈ5ı?IHɵ͍L
(443) NMR e[q5EG̰ȍ#+. (4). NMR 5EHˏŧͪċDžþłŧ7ȟͤȸů. öΜ#+ˏŧͪċDžþöʈ5/0
(444) Īʵ NMR e[q7ɖůL̞.+ 1H-NMR e[qE: 1ț˘#+2. 13. C-NMR e[q7]Wc~qÙLe[qpje. J
(445) Z;
(446) §;E: D-\_e7]Wc~qÙ)I*I. ̉#+νpj8ʪ#04ξ %4K-
(447) ˏŧͪċDžþöʈ58Z;
(448) §;E: D-\_eı?I0+. (5). SST 5H§;7ů. SST 5H§;7ŨňL̾<H+B
(449) bQ[kQg[cLʃ0 ÷Gõ#+ SST ʖ5ı?IH§;Lɖů#+ UVJ ò˝÷ɳLΫƴΉ1̮Ż% H2 SST E: UVJ ˎ̂ʖ̮Ż"I++BνFig. 2-4, Bξ
(450) SST 7ĵŃ5b Lɭž#å}gmqLʃ0 SST LŁĠ#+νFig. 2-4, Cξ ĭȠ5#0 UVJ ˎ̂ ʖCŁĠ#
(451) IF7NJõɵ5ı?IH§;ɦƢLɖů#+2. J
(452) SST ʖNJõɵ7. §;ɦƢ8˓ 13 mM 1G
(453) UVJ ˎ̂ʖNJõɵ7§;ɦƢν˓ 3 mMξ2Ȳ͗#0 ȁƼ5εÙLʪ#+νFig. 2-4, Aξ. (6). ĖΜ#+ SST 7§;ʂʁ˻. Ǘġ#+ UVJ L_^shôɿ#űÈΫƴΉ1̮Ż#+2. J
(454) ĖΜ"I+ SST. ̮Ż"I+νFig. 2-5, Aξ ĖΜ#+ SST L HBSS 5ŁĠ#
(455) 39˚C 1QYq #4F§;ɦƢLɖů%H2
(456) ŎΰɌ¤7§;˜Ƕʔ5ŖĈ#+νFig. 2-5, Bξ 2-DOG Ɍ¤1 SST LQYq#̰ː˒7ͽ˖LΓŴ%H2
(457) §;ɦƢ7dz8. 21.
(458) Džþ"I+νFig. 2-5, Bξ HBSS ¤1 1 ǶΎŎΰ#+˱7§;ɦƢL 100%2%H2
(459) 2-DOG Ɍ¤1Ŏΰ#+˱7§;ɦƢ8ȁƼ5ÆÙLʪ#+ν˓ 20%ξ ?+
(460) 2-pV Yc\_e2άÄ7ȟͤ1G̰ː˒7ͽ˖LΓŴ#4 2-pVYcX[q eLıA 2-DOGal Ɍ¤1Ŏΰ#+˱7§;ɦƢ8 2-DOG ˱EGȁƼ5ε
(461) HBSS ˱ 27Ύ5ȁƼƖ8@FI4.+νFig. 2-5, Cξ. (7). SST 5H MCT ͵Áŧ7ʑɽE:ƃň. _^shôɿ5EGĖΜ#+ SST F RNA LNJõ#
(462) RT-PCR 5EG MCT1, MCT2, MCT3 E: MCT4 ͵Áŧ7ʑɽ̰ȍL̞.+2. J
(463) MCT2 E: MCT4 7. 5£˵2C˓ 900 bp 7yr̮Ż"I+νFig. 2-6ξ ͕͠îL̞.04˱ν π ξ18&ICyr8@FI4.+ in situ xQkQhc5EG MCT4 mRNA 7ƃňLʨ̹#+2. J
(464) Ol. geLʃ+˱50c\sțõ"I+νFig. 2-7, Aξ
(465) ge Lʃ+˱18c\s8țõ"I4.+νFig. 2-7, Bξ űÈΫƴΉ1 X ˨~ PE:eQrL̮Ż%H2
(466) SST ʖE: UVJ ˎ̂ʖ5c\s@F I+νFig. 2-7, C E: Dξ. (8). RNA-Seq Lʃ+ SST ʖ2 UVJ ˎ̂ʖ7͵ÁŧʑɽƖ̰ȍ. SST ś7§;Lʂʁ%HWtfLªɖ%H+B
(467) RNA-Seq Lʃ+͵Áŧ ʑɽ7ˤ˯ʔ̰ȍL̞.+ µŁ7űγ1ƱFI+pjgmq8
(468) DDBJ Sequence Read Archiveνhttp://trace.ddbj.nig.ac.jp/dra/index.htmlξ5ʒ·#+νʒ·ʌĨπDRA003919ξ YiYshDe~[qYsh
(469) }|;p{r^shYsh νPDKξ
(470) §;˾ȵ˖ͽ˖νLDHξ2.+̰ː˒7ͽ˖7ʑɽ8
(471) SST 2 UVJ ˎ̂ ʖ27Ύ1ȁƼƖ8@FI&
(472) \_eqejνGLUTξ5/0Cȁ. 22.
(473) ƼƖ8țõ"I4.+νFig. 2-8ξ SST qe[q7pjgmqLʃ 0 KEGG PATHWAY m}\̰ȍνhttp://www.genome.jp/kegg/pathway.htmlξ
(474) E: SST 2 UVJ ˎ̂ʖ7 GO gmqLʃ+͵ÁŧʑɽƖ̰ȍL̞.+
(475) ̰ː˒7° ͧLʪ%˝ȏ8ƱFI4.+νTable 1ξ ǰ
(476) SST 50
(477) Mě͵Áŧ1H c-Fos E: c-Jun 7ʑɽ UVJ ˎ̂ʖ2Ȳ͗#0̔#dz#0H. 2K.. +νFig. 2-8ξ. (9). UVJ 5H;˖Î˞. RNA-Seq ̰ȍ5EG SST Æ;˖ɷƾ5I0HĦ˻ƹʪļ"I++B
(478) Æ ; ˖ 7 ˛ ˬ 5 ̕ ʷ % H HypoxiprobeTM-1 L ʃ 0 SST 7 ; ˖ Î ˞ L ̾ < + HypoxiprobeTM-1 LΠ˽ɀž#+˱18
(479) SST ʖ5 HypoxiprobeTM-1 7c\s̮Ż "I+
(480) UVJ ˎ̂ʖ508c\s̮Ż"I4.+νFig. 2-9, Aξ ?+
(481) ɛť7@Lɀž#+_q˱18
(482) SST ʖ
(483) UVJ ˎ̂ʖ7&ICc\s 8@FI4.+νFig. 2-9, Bξ %4K-
(484) SST ʖ8Æ;˖ɷƾ5I0H. 2. K.+. (10) SST ʖ˙˺7q_rOɅƹ SST ʖ7q_rOɅƹL̾<H+B
(485) JC-1 Lʃ0 UVJ LȒ̍#+ SST ʖ8˦̛̍åƨ
(486) ͏̛̍å8=2M3@FI4.+
(487) UVJ ˎ̂ʖ8͏̛̍ åƨ̮Ż"I+νFig. 2-10, Aξ CCCP LɑĈ%H2
(488) SST ʖ
(489) UVJ ˎ̂ʖ2 C5͏̛̍å=2M3@FI44.+νFig. 2-10, Bξ Image J Lʃ0 530 nmν˦ ̍ξE: 590 nmν͏̍ξ5H̛åƨƢLůď#
(490) FI ȲLˈõ#+2. J
(491) SST. ʖ7 FI Ȳ8 UVJ ˎ̂ʖ2Ȳ͗#0ȁƼ5Æ
(492) %4K-q_rOɅƹÆ. 23.
(493) . 2ʪ"I+νFig. 2-10, Cξ CCCP LĈ+27 SST ʖE: UVJ ˎ̂ʖ. 7 FI Ȳ8 CCCP LɑĈ#04 SST ʖ7 FI Ȳ2Ȳ͗#0ȁƼƖ8@FI4.+. 24.
(494) Absorbance at 280 nm (A.U.). 300 250 200 150 100 50 0 20. 40. 60. 80. 100. 120. 140. 160. Elution vol. (ml).
(495) à àq½ÉëĎÞ¢éĐą÷êČÿãĒÐÙÜOjѬ¹ Oj
(496) ຠæČćē
(497) Ĕàq½ÉëĎh¢ÐÙÛrÆÉ»fa à&Æ .ÆÉÑʺon´2^ē ĔÐfƺ5¡!âõðäÐÆÉ»on`HàS ÇÜràûĒÍ|ÆÉ».
(498) A. B Motility score. 4 3 2. *. 1 0 H2O 1. 2. 3. 4. 5. 6. 7. 8. 9. 10 11. Fraction No..
(499) yæČćàq½É ÐÙÜ OjѬ¹ on`Hrrà yæČćͬÆÉ»&ÆÉràûĒÀÙÓQ6Í|ÆÉ» rÂ5Ñ¡!пÜD²àÕÉ»(rà!+×Ð=5à"
(500) ÎÏÜÙ¾cƺ5Ñ¡!HàÑ\«ÍÆÉ»rÀÙÓÐon` HÂØÚÝÉ»Ð<ÆÌSI?¼Ûē#Ĕ».
(501) S 1 2. 3 4. 5 6 7. 8 9. 10 11 12 13Fr. #. 1 4 4. 4 4. 4 4 1. 1 4. 4 4. 4 4 motility score.
(502) Ð ÐÙÜOjѬ¹ on`HrÑ TàËÉ»Ñ(rà ÐÆºčđĉčĀöđ¥ÐÙÛ,ÆÉ»z $Í|ÆÉrк5Ð<ÇÜC½¡!L. UÂ;ÅÝÉ»ÑrÀÙÓē
(503) . Ĕà)ƺÐÙÛ¬ÆÉ»(ràcÆÉRÑ5Ñ¡!Hà\Ð|ÆÉ»z$Òo n`HjÑïĄõ÷à|ÆÌ½Ü»
(504) rÀÙÓÑb)j».
(505) A18. B. Lactic acid (mM). 15 12 9. C. 6. *. 3 0 SST s. Epithelial cell.
(506) Ð ÐÀÄÜ¥Ñ9§¹ ÑiàKƺďĒíĒąäéĐóäðéîċđàq½Ì wÖÉÒ w ē± wĔàÛÆÌ¥à9§ÆÉ» wÀÙÓ± wÑOjÐ+ÖÝÜ ¥à9§ÆÉ»:¶à-½ºA0¸Í|ÆÉ» wÑ¥§Ò± wÑ ¥§Î]ÆÌSIзÁËÉēĔ»ďĒíĒąäéĐóäðéîċđÐÙÛ à ÛÇēĔÎFēĔÑ wià|ÆÉē
(507) μ Ĕ».
(508) Lactic acid (μmol/mg protein). B. A . Arbitary unit % of control. C. 3 Glc 2-DOG 2. *. 120. *. 0 0. 140. *. 1. 30 60 90 120 Incubation time (min). a. a. 100 80 60. b. 40 20 0 Glc. 2-DOG. 2-DOGal.
(509) Ñ Ñ#¬ÀÙÓ¥poe9¹
(510)
(511) àìČëùĒñnÐÙÛ#¬ÆºÍäđèĊĂĒ÷ÆÏÂÚRvÐ¥§àe9ÆÉ» ©ÑìČëùĒñnÐÙÛ
(512)
(513) Â#¬ÅÝÉēμĔ»#¬ÆÉ
(514)
(515) à êĎìĒïēĔÖÉÒ öåèîêĎìĒïē Ĕà+×
(516)
(517) Í1µÆº1µdÑ ¥gBàRvÐ9§ÆÉ»:¶à-½º¥gBàA0
(518) ÍÆÉ»*R©Ñ Ð<ÆÌSI?¼Ûē Ĕ»#¬ÆÉ
(519)
(520) à êĎìĒïēĔº öåèî êĎìĒïē ĔÖÉÒöåèîçČé÷Ēïē Ĕà+×
(521)
(522) Í1µÆºR©FÑ ¥gBàe9ÆÉ»:¶à-½º¥gBàA0
(523) ÍÆÉēs'©ÍSI?¼Û ē ĔĔ».
(524) MCT1. MCT2. MCT3. MCT4. − + − + − + − +. S17 M. − +. £ £
(525) 5ÑumT¹
(526) ÁÚ àOƺ àËÉÑʺºººÀÙÓ ÑksvāČ äąĒàq½Ìà3@ÆÉ» ēďĒđĕĔÒº
(527) ÁÚ àOÆÉÑÊ à ßȺ*YÐ àËÉ»-Ñ:¶à½º vÏUà|ÆÉ»5§ąĒæĒ» ÀÙÓÑďĒđÐ ÑûđøÂ{ÅÝÉ».
(528) A. B. C. D D.
(529) Ð ÐÀÄÜ Ñum¹ ÑiàKƺZāĐĒĀàq½É
(530)
(531) úäĀčóäñĒîċđÐÙÛ Ñ>/à{ÆÉ»-Ñ:¶à½º vÏUà|ÆÉ»âđôðđïāĐĒĀàq ½ÌúäĀčóäñĒîċđàËÉ»ðđïāĐĒĀàq½ÌúäĀčóäñĒîċđàËÉ» ïČäøà ÿãĎćЮƺ:³G¨Í;ÆÉ»ïČäøà:³G¨Í;ÆÉ ē μ Ĕ».
(532) ΠαwÐÀÄÜ£
(533) 5Ñum?T¹ α wÐÀÄÜ£
(534) 5um?TàËÉ»ăõéïāĐõ÷ÒºÂáºþĎýđ¥ öüøĐëùĒñèùĒñÀÙÓ¥^¤ē
(535) $ ĔººÀÙÓêĎìĒï ÷ČđïĄĒòĒē ĔЪÇÜ£
(536) 5Ñum§ē &%"
(537) '&#ĔàÆÌ½Ü»xÒº ± wēĔÎ ÐÀÄÜ£
(538) 5Ñum?à<Q4PÆÌ|ÆÌ½Üē Ĕ» ē
(539) ĔÐÙÛ Î± wÑ à]ÆÉ» !£
(540) 5Qº!ïìâº!» ÐÀ½Ìº%Âá£
(541) 5ÑumÂÆÃ3ÆÌ½É».
(542) A . B.
(543) ÐÀÄÜ¥¹ t"8VÐÙÛº¥lJÑ àXÆÉ» à°_=ÆÉ ĈïÑ
(544) »on´2^à°_=ÆÉĈïÑ
(545) »NNÑîêùĎàͺ Wà VÐÙÛ¯Í|ÆÉ». àz$ͺ±. wà ¦z$Í|ÆÉē μĔ» MÐÀ½Ìº. ÔÑ~Â;ÅÝÉ».
(546) A. B. C 1 FI ratio (590/530 nm) . a. −CCCP. 0.8. +CCCP. 0.6. b. 0.4. b. b. 0.2 0 Epi. SSTs. ÀÙÓ±wÑĆ÷ìđøčâ`H¹ VÐÙÛºE Ñ7/ēĔÖÉÒ±7/ēĔÐÀÄܺ
(547) ÀÙÓ±
(548) wÑ Ć÷ìđøčâ àÕÉ»
(549) àz$ͺ±
(550) wà ¦z$Í|ÆÉē Ĕ»
(551) ē[ĔÀÙÓ±
(552) wēĔÐÀÄÜÑ]ēĔàe9ÆÉ»¹ -Ñ:¶à½ºA0¸! Í|ÆÉēs'©ÐSI?¼Ûē ĔĔ»¹ ±7/ē ĔÍÒ±
(553) wÎ]ÆÌ
(554) ÑĆ÷ìđøčâ`HÒSIÐú¹ àcÇÜÎē Ĕ±
(555) wÑĆ÷ìđøčâ`HÒ
(556) Î*}BÖÍÆÉ» .
(557) GO:0004672. GO:0005622. GO:0017048. GO:0006468. GO:0016459. GO:0003774. GO:0007018. GO:0003777. GO:0008289. GO:0005524. GO:0006820. GO:0005576. GO:0006412. GO:0003735. GO:0005840. GO:0005515. GO_id. MF calcium channel activity. MF protein kinase activity. CC intracellular. MF Rho GTPase binding. BP protein phosphorylation. CC myosin complex. MF motor activity. BP microtubule-based movement. MF microtubule motor activity. MF lipid binding. MF ATP binding. BP anion transport. CC extracellular region. BP translation. MF structural constituent of ribosome. CC ribosome. MF protein binding. GO_name. 10. 415. 367. 12. 423. 58. 58. 39. 38. 21. 929. 20. 110. 99. 99. 98. 2354. Number of sequences. -4.269214999. -4.332896983. 4.357143873. -4.468511328. -4.718625472. -4.804414293. -4.804414293. -4.827499659. -4.870426596. 5.334001178. -5.361703161. -5.426567452. 6.049948334. 6.373236731. 6.527433457. 6.548828012. -7.794049752. Z score. 1.96162E-05. 1.4716E-05. 1.31771E-05. 7.87658E-06. 2.37444E-06. 1.55205E-06. 1.55205E-06. 1.38258E-06. 1.11358E-06. 9.60719E-08. 8.24409E-08. 5.74481E-08. 1.44892E-09. 1.8508E-10. 6.69063E-11. 5.79903E-11. 6.43929E-15. P-value. 0.000306936. 0.000244654. 0.000233674. 0.000149655. 4.86E-05. 3.44E-05. 3.44E-05. 3.44E-05. 3.29E-05. 3.19E-06. 3.13E-06. 2.55E-06. 7.71E-08. 1.23E-08. 5.93E-09. 5.93E-09. 1.71E-12. False discovery rate. "#'$
(558) . GO:0005262.
(559) GO:0005198. GO:0016791. GO:0031683. GO:0019001. GO:0005509. GO:0008146. GO:0008236. GO:0005179. GO:0004871. GO:0004867. GO:0015074. GO:0008017. GO:0005737. GO:0030036. GO:0004198. GO:0016311. GO:0005856. GO:0070588. GO:0005882. GO:0007156. MF structural molecule activity. MF phosphatase activity. MF G-protein beta/gamma-subunit complex binding. MF guanyl nucleotide binding. MF calcium ion binding. MF sulfotransferase activity. MF serine-type peptidase activity. MF hormone activity. MF signal transducer activity. MF serine-type endopeptidase inhibitor activity. BP DNA integration. MF microtubule binding. CC cytoplasm. BP actin cytoskeleton organization. MF calcium-dependent cysteine-type endopeptidase activity. BP dephosphorylation. CC cytoskeleton. BP calcium ion transmembrane transport. CC intermediate filament. BP homophilic cell adhesion via plasma membrane adhesion. 60. 25. 15. 15. 269. 34. 21. 13. 46. 18. 13. 36. 221. 18. 13. 20. 42. 10. 31. 53. 2.801202112. -2.831161577. -2.853316217. -2.853316217. -2.864577588. 2.892907378. 2.976241845. 3.000344938. -3.065659791. 3.072754654. -3.091820668. -3.130481343. 3.149034063. -3.152004905. -3.162832965. -3.210644436. -3.603256105. -4.036394977. 4.116758099. -4.232014107. 0.005091262. 0.004637929. 0.004326555. 0.004326555. 0.004175658. 0.003816938. 0.002918046. 0.00269674. 0.002171903. 0.002120928. 0.00198933. 0.001745201. 0.001638111. 0.001621535. 0.00156242. 0.001324377. 0.000314256. 5.42788E-05. 3.84239E-05. 2.31608E-05. 0.035857906. 0.034269142. 0.032881818. 0.032881818. 0.032881818. 0.031728297. 0.025038717. 0.023911095. 0.019921593. 0.019921593. 0.019598584. 0.017854749. 0.017429501. 0.017429501. 0.017429501. 0.016012922. 0.003980576. 0.000721908. 0.000537935. 0.000342265.
Outline
関連したドキュメント
In addition, more than 50% of fluorescence positive cells exhibited shrinkage and rounding even in the absence of anti-Fas antibodies (about 56, 65, and 56% of PKR-, dN-,
however, expression of OCTN2 on apical membranes of intestinal epithelial cells was.. reduced in pdzk1 -/- mice, compared with wild-type mice, with a concomitant
Pim-3, a proto-oncogene with serine ⁄ threonine kinase activity, is aberrantly expressed in human pancreatic cancer and phosphorylates Bad to block Bad-mediated apoptosis in
熱力学計算によれば、この地下水中において安定なのは FeSe 2 (cr)で、Se 濃度はこの固相の 溶解度である 10 -9 ~10 -8 mol dm
For staggered entry, the Cox frailty model, and in Markov renewal process/semi-Markov models (see e.g. Andersen et al., 1993, Chapters IX and X, for references on this work),
A lemma of considerable generality is proved from which one can obtain inequali- ties of Popoviciu’s type involving norms in a Banach space and Gram determinants.. Key words
Given a compact Hausdorff topological group G, we denote by O(G) the dense Hopf ∗-subalgebra of the commutative C ∗ -algebra C(G) spanned by the matrix coefficients of
Here we shall supply proofs for the estimates of some relevant arithmetic functions that are well-known in the number field case but not necessarily so in our function field case..