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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

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(1)Title. 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( 本 文(Fulltext) ). Author(s). 松崎, 芽衣. Report No.(Doctoral Degree). 博士(農学) 甲第666号. Issue Date. 2017-03-13. Type. 博士論文. Version. ETD. URL. http://hdl.handle.net/20.500.12099/56214. ※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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(7)  F¿Ž|{ 9. –B˜ ¤´. 10. –G˜ vsUt€ (1). `ÄR‰. (2). ±¬. (3). UVJ nN‰ ³q. (4). ›] pV»R)$. (5). =ˆ¼?6,;3A. (6). ʃK?6,;3AÆHPLCÇ. (7). OV?6,;3AÆPLCÇ«d?6,;3A ÆTLCÇ. (8). NMR. (9). SST F¿_À. (10) SST TÁ (11) SST F¿Ž¨ _ (12) RT-PCR (13) in situ 04<(%A. :@. (14) RNA-Seq  cDNA ;4;< {™ (15) RNA-Seqde novo $@4<+3>@. 9=¯x. (16) ž¥J¿œ zN (17) 7,@-<‚k _ (18) ¢°¯x 20. –C˜  y (1). =ˆ¼?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¿Ž¨. (7). SST

(14)  MCT ¾I] ’‹c[. (8). RNA-Seq  SST D“ UVJ š©D“ ¾I]’‹e¯x. (9). UVJ

(15) ¿œL¡. (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. .=I½Ÿ·. –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) ›]W†S‰ 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 ›] §Œ

(24) EiÃ. (3). PKC

(25) ›]'@2 <@¿S. (4). ›]

(26) ^[  PKC &3A8 X_. –Y˜ ¦a. 53. –H˜ bo. 57. –H• £o. 58. µ¸. 64. hrŠ. 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.

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(98) öȐ?+8ŸöȐ7ˉ1HνBobr et al., 1964ξ ›Ƣ7± Ƃ1Ñç#+ˏŧ͈̖"IHȂΎ8ʵ5E.0ʍ4.0H clŠ—lR νMeleagris gallopavoξ18

(99) 815 ͦΎ

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(182) UVJ 7NJõɵLɑĈ#+ŎΰɌ1ˏŧLQ—YŽ‚™q%H2

(183). ˏŧ7ͪċDžþ"I

(184) ˏŧ7ͪċˡˢǶΎƣ΋%H νMatsuzaki et al., 2014ξ Ʌƹɵ͍ı?IH. 2ǴF24.0H. I8

(185) UVJ 7NJõɵ5ˏŧ7ͪċLƞ"'HȄʢ7ʁɿ. 2Lʪļ%HC71H ). 1Ȇʤʹ18

(186) SST 8ˏŧ7ͪċ. LŸɅƹď%Hɵ͍ı?I0H2½̼Lʽ0

(187) 7ɵ͍Lĭů#

(188) )7Êʃȣ ƠL̰Ǵ%H. 2Lʚʔ2#+. 8.

(189) ˁ¬ʾ ˏŧ͈̖ˉʅȋˏŧͪċDžþłŧ7ĭůE:ˏŧ͈̖ˉ5H §;ʂʁȣȟ. ˁ›ˊ ˧̿  ηάLıAś7ÈìĢˏċɵ18

(190) ±Ƃ2Ģˏ7jQˆ—\8Ʒ&#C›̉'&

(191) ˏŧ2ęLͲ÷4ǶȂ2Œǂ5êŨ"'H+B7̾ˊȣȟŨň%H 7ǶΎ̾Ǫ7 ƬąLNj.0H7͈ˏ1G

(192) ͈ˏL̞ċɵ18žˏƯ7ˏŧ8ęˉì1›ůȂ Ύ͈̖"I

(193) ǖę?17Ύ

(194) Èɕº͛1CĢˏ˻LÔ.+??ˣǐ"IH ͈ˏ7ȂΎ 8ʵ5E.0Ƞ1

(195) Í9ŵʮ508

(196) Rf‘1 11 DZνSittman and Abplanalp, 1965ξ

(197) t–q’1 35 DZνNalbandov and Card, 1943ξ

(198) clŠ—lR1 117 DZνChristensen and Bagley, 1989ξ2őĴ"I0H ηά5H͈ˏ7Œ8

(199) ęˉ7ŧų̃ʳ̞͹ νutero-vaginal junction; UVJξE:ɞǬ͹5ƃň%Hˏŧ͈̖ˉνsperm-storage tubules; SSTξ1HνBobr et al., 1964ξ žˏƯ7ˏŧ8 SST 5Ñç#+7-ͪċLÚȦ#

(200) Ģˏ˻LÔ.+??Èɕº͛1΋ȂΎˣǐ"IHνBakst, 1987ξ ɶ˃%<8

(201) ˏŧ 8͘ęˉ7¤1ΠȦɷƾFͪċɷƾ;7÷GǿL 2 ŁνžˏƯE: SST F7 ǦõƯξ̞. 21H Rf‘18

(202) SST FǦõ"I+ˏŧ8 UVJ 7ˎ̂ʖ˙. ˺FǦõ"IH heat shock protein 70 5EGͪċLíɅƹď%HĦ˻ƹʪļ"I0 HνHiyama et al., 2013ξ

(203) )7¹7ͬʴ50ˏŧ7ͪċLþƲ%H̩ł8ŸǴ 1H . I?15

(204) SST 5Hˏŧ7ͪċÚȦ7ʁɿũʔƬą˴Ż"I0H. νFroman, 2003; Froman and Feltmann, 2005; Froman and Feltmann, 2010ξ

(205) SST ͈̖ˏŧ5 HͪċÚȦ7öŧŠWtf‰5/08ǴF54.04 Ȇʤʹ18

(206) SST 8ˏŧ7ͪċLŸɅƹď%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 7Ȋ¾ž1ί˹#+ α8

(212) ‹qYF͎ ç#+ǀιʃίǭLŸǮ˞α1 

(213) ȵ8ȵͭȵLŸǮ˞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[w•d ™fνɗđξF͎ç#+ HBSS 10 ×ɌνW“cR‰

(218) ‡\vcR‰

(219) ɩ;ȵ˖sq ’R‰

(220) ~Sw™“”mrŸıξ5

(221) ā͝7ɦƢ54HE Milli-Q ȵ1ƘͿ#

(222) CaCl2, MgSO4•7H2O, NaHCO3 E:~Sw™“”mrLɑĈ#0Ìʃ#+. (3). UVJ NJõɵ7̦̾.  ͥʂŠeRf‘LǮΨƇȮ5EGūȞȨ"'+7-

(223) ͘ęˉLǞõ#+ ͘ęˉLŧ ų͹F̃͹50÷΍#

(224) ŧų͹2̃͹27ŕʉ5Åˮ%H UVJ LǗġ#+ UVJ LʁɿήŔȵν0.9% NaClξ¤5ġG

(225) SST Lı?4͹ÅLűÈΫƴΉž1ΕĜ#+ q’ˆ—\#+ UVJ LTmƒ—r“~lŽ™5çI

(226) 500 mg ˛ˬ/ml 54HEȶ ð PBS LĈ+Ư

(227) ̰Ăyaˆ1˙÷#

(228) ͲŰ “om[eˆYa™1ǣnj#4F ȶ1 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-q™‡̝ɾ̳ˈʘLʃ0ˏŧɦƢLˈõ#

(233) ɦƢ 2.0 × 107 sperm/ml 54HE HBSS 1ƘͿ#0OmgQ5ʃ+  ˏŧ7ͪċƹL̷Ð%H+B5ˏŧͪċOmgQL̞.+ UVJ NJõɵL 10%Ŷ΂ ıA HBSS 7r•m€L€‘elm[cŒ™”5Êǀ#

(234) Ɇċz‘~P—1Žç#+ Êǀ#+r•m€8

(235) OmgQ5ʃHā5 39C 1ªɕ#+ ͝7ǰȾ1Ǘġ#+ ˏŧƿɥɌLr•m€7 20%Ŷ΂ĈνȀ˚ˏŧɦƢ 4.0 × 106 sperm/mlξ

(236) „mq€” ™q1 39 ˚C 5Ô-4FűÈΫƴΉνM165FCρ‘QW‡Q[•ceo‰f

(237) ǯŷ đξž1̮Ż#+ 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\‘~P™5Î%H UVJ NJõɵ8

(244) Z;O—‹tR‰ym~N™ ν20 mM Z;O—‹tR‰

(245) pH 7.4ξLʃ͝7ǰȾ1̦̾#+ UVJ NJõɵ8 Superdex 200pg νGE “e]O

(246) DZ΁ƗξW‘‰ν⌀ 1.6 × 75 cmξLʃ0öΜ#+ ˾ȴ#+Z;O—‹tR‰ym~N™240 mlν2 W‘‰Ŷʷöξ1W‘‰LƜ̢ď#+ 7-

(247) UVJ NJõɵ 1 ml LW‘‰5O€‘Q#

(248) "F5Z;O—‹tR‰ym~N™140 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% Ŷ΂ɑĈ#0ͪċOmgQ5. EHe[’™t—\L̞

(253) 24 ǶΎƯ7ͪċƹε.+öʈLΚBī›#+ Superdex 200pg 7 QrŶ΂ (V0) E:öŧ΂Lˈõ%H+B7ȡɚǽ˨8

(254) Ferritin (440 kDa)

(255) Aldolase (158 kDa)

(256) Conalbumin (75 kDa)E: Ovalbumin (43 kDa)7ɛõÈ ʷ5EGÊǀ#+. (6). εͣɌÈ[•‡q\‘~P™νHPLCξ.  UVJ NJõɵ7 Superdex 200pg ÆöŧöʈνɛõÈʷ 100110mlξLò˝¨ɰ#+7 - Milli-Q ȵ5íɛ̰%H. 21ɦ˪#

(257) C-22 W‘‰νNavi C22-5ρ4.0 × 150 mm, ĸå. ˔̘ξLʃ+ HPLCνƎɄ̦Êǂ

(258) ²ͺƗξ5EGöΜ#+ ʳċʝ A L 0.1% q’ ~“V•ͼ;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 7’tO\‘dT—q1ɛõ#

(262) ȀƯ5 A : B = 100 : 0 1 5 öΎɛõ#+ 220 nm 5 HIJåLțõ#

(263) ”_™k™1̴·%H22C5

(264) }™[̮Ż"I+ɛɌLöġ# + ƱFI+öʈ8ò˝¨ɰ%H. 25EG TFA LΕĜ#

(265) Milli-Q ȵ5íɛ̰#+7. -

(266) HBSS 5 1% Ŷ΂ɑĈ#

(267) ͪċOmgQ5EHe[’™t—\L̞.+. (7). öġ[•‡q\‘~P™νPLCξE:̗ƈ[•‡q\‘~P™νTLCξ.  PLC 8

(268) PLC X‘e€”™q c’W^“ 60 F254νŅůƈĚ 1 mmρŠ“[

(269) ʚλđξ Lʃ0̞.+ YŒ}‘’Lʃ0€”™q5a—€“Le†mq#

(270) ͼ;Tl“ : Šjw™“ : H2O = 6 : 4: 1 7ɛť1ƅ΍#+7-

(271) €”™qLpc]™j™¤1¨ɰ" '+ a—€“Le†mq#+ÅˮF 1cm !27đʈ5ö

(272) €”™qFc’W ^“LǚġGX‘e̸γˉ5öġ#+ c’W^“5ƅ΍ɛťLĈ

(273) ɤ#ǣnj# 4F 30 öΎNJõ#+Ư

(274) ɛťLɧͬ#

(275) ʼ˖XeLijºH. 12. 25EGɛťLʊ.

(276) Ĝ#+ ˝ǹ?+8VQ“ɷ7ɌÈL 10 μl 7 Milli-Q 5ɛ̰#+7-

(277) HBSS 5 1%Ŷ ΂ɑĈ#

(278) ͪċOmgQ5EHe[’™t—\L̞.+  PLC 5EGƱFI+öʈ5ż#

(279) TLC öȍL̞.+ YŒ}‘’Lʃ0 TLC O“ ˆc™q c’W^“ 60 F254νŠ“[

(280) ʚλđξ5a—€“Le†mq#

(281) ͼ;Tl“ : Šjw™“ : H2O = 6 : 4: 1 7ɛť1ƅ΍#+ ƅ΍Ư

(282) €”™qLέ¨#

(283) ’—‹’ p—;ʑ̸̘̍ν5% 12MoO3ςH3PO4, 100%Tjw™“ɛɌξ5€”™qLɊɟ#+7-

(284) „mq€”™q1Ĉɯ%H. (8). 25EGʑ̍"'e†mqL̮Ż#+. NMR.  1H-NMR

(285) 13C-NMR

(286) E:¬ȥã NMR eƒ[q“νHMQC E: HMBCξ8

(287) JEOL GSX-500 Ȗʩȴêθ̤ˮν500 MHzρDZȆΞŧ

(288) ǵƎƗξLʃ

(289) ΀ȵνD2Oξ5ɛ̰ #+̸ǭLɖů#+ ƱFI+eƒ[q“p™jL̰ȍ#+7-

(290) L-§;νc\‡O“ r’mlξ7eƒ[q“p™j2ɭī#0›̉%H. (9). 2Lʨ̹#

(291) ďīɵLĭů#+. SST 7§;ů΂.  ͥʂŠeLǮΨƇȮ5EGūȞȨ"'

(292) ͘ęˉLǞõ# UVJ LǗġ#+ Ǘġ#+ UVJ ˎ̂L OCT _—zR—rνa[‘~NQ—om[dŒz—

(293) ¤Şđξ5ĎŌ#

(294) 80˚C 1ò˝#+7-

(295) [’VejmqLʃ0Ě" 30 μm 7ò˝÷ɳLÊǀ#+ ” ™b™‡Q[•kQg[c—νPALM CombiSystemρW™“nNQe

(296) ǯŷđξLʃ 0 UVJ ò˝÷ɳF SST ?+8 UVJ ʖ˙˺L÷Gõ%22C5

(297) ”™b™Lɭž #+΢ʷ2ò˝÷ɳ7Ě"F

(298) ÷Gõ#+˙˺œ7ÈʷLˈõ#+ ŁĠ#+˙˺œ 5 100 μl 7 Milli-Q ȵLɑĈ#0

(299) †’q•—„‹dsQb™Lʃ0„‹dsQf# +7-

(300) „‹dsQf#+ɛɌL 20,400 × g

(301) 4˚C 1 10 öΎͰƶöΜ#

(302) Ty†”™j Lʃ0ɒL¨Ņ#+ Milli-Q ȵLĈ0íɛ̰#+Ư

(303) L-Lactate Assay KitνCayman

(304). 13.

(305) ˆcX—ƒξLʃ

(306) ̼ǴǾ5ư.0§;ů΂L̞.+ %4K-

(307) ͽ˖ɛɌ

(308) ̥ͽ˖ ɛɌ

(309) ̛åŏ͍LıAOmgQym~N™5a—€“ɛɌLɑĈ#

(310) 20 öΎŲɕ1ğ Ƹ"'+ r‘‹—rYŒ}‘’1ğƸɌ 4 μl L̳G2G

(311) ̛åΫƴΉνBX51ξE : Image JνOŠ’Wņʽ̠ʁʤʹǂ

(312) Š’™‘—rƒξLʃ0r‘‹—rYŒ}‘ ’ì7ğƸɌʑ%H̛åLů΂#+ ”™b™‡Q[•kQg[c—5EG÷Gõ #+˙˺œ7ÈʷF

(313) SST ?+8 UVJ ʖ˙˺5H L-§;ɦƢLˈõ#+. (10) SST 7ĖΜ  SST 7ĖΜ8

(314) _‘^s™hôɿLʃ+ǰȾνKing et al., 1999ξ5EG̞.+ ͥ ʂŠeLǮΨƇȮ5EGūȞȨ"'

(315) ͘ęˉLǞõ# UVJ LǗġ#+ űÈΫƴΉ νM165FCξž1 SST Ũň%H͹ÅLq’ˆ—\#+7-

(316) SST LıA UVJ ˎ̂L lŽ™5Ǘġ#+ _‘^s™hɛɌν10 mg/ml _‘^s™hνĸå˔̘ƔȜξL ıA HBSSξL 1 mg ˛ˬ/μl ɑĈ#̰Ăyaˆ1˙÷#+7-

(317) _‘^s™hɛɌL" F5 500 μl Ĉ

(318) ǣnj#4F 37C 1 20 öΎQ—YŽ‚™q#+ _‘^s™hô ɿƯ

(319) ȶð#+ğƸÚȦɌν10 mg/ml ɴ̝ɒO“ˆ—LıA Ca2+/Mg2+Ÿı HBSSξ L 5 ml ɑĈ#0ͽ˖ğƸLÚȦ"'+ ğƸɵLcŒ™”5ʳ#

(320) ‡Q[•}ƒmq LʃűÈΫƴΉž1ĖΜ"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\“_™eŸı HBSSξ?+8 2-DOGal Ɍν1 mM 2-pVYcX‘[q™eLıA\“_™eŸı HBSSξ¤50 39C 1Ŏ. 14.

(325) ΰ#+Ư

(326) Πˮ#0 SST Lȹɣ"'

(327) ŎΰɒLŁĠ#+ ŎΰɒL 20,400 × g

(328) 4˚C 1 10 öΎͰƶöΜ#

(329) ɒ5ı?IH L-§;ɦƢL

(330) L-Lactate Assay Kit νCayman

(331) ˆcX—ƒξLʃ0͝7ǰȾ1ů΂#+. (12) RT-PCR  RNAisoνjW‘yQV

(332) ̏ɄƗξLʃ

(333) _‘^s™hôɿ5EGĖΜ#+ SST  Fè RNA LNJõ#+ ɐç#+ DNA L DNase IνjW‘yQVξ5EGö̰#+Ư

(334) ReverTra AseνȌɂ˗

(335) ŜΑƗξLʃ0͕͠îğƸL̞.+ cDNA LΆŋ2#

(336) MCT1 νg—eπ5 - GAACCCTGCCTTAACCATGA - 3

(337) O—lg—eπ5 - GGCACA CCCCATTGTAAATC- 3 ξ

(338) MCT2νg—eπ 5 - TGTGGCTGGGTCACTTATGA - 3

(339) O—lg—eπ 5 - CTGGCCACAATCACAATCAC- 3 ξ

(340) MCT3ν5 - CTGGGTA TGGCGTTGAACTT - 3

(341) O—lg—eπ5 - TAAGTAGGACCAGCCCGATG- 3 ξ? +8 MCT4νg—eπ5 - AGCACCAAGCCAATAGCACT - 3

(342) O—lg—eπ 5 GGGTCTGGCACTCAACTTTC- 3 ξ7ɶʍʔ€‘Q‡™Lʃ0 PCR L̞.+ ì͹ ȡɚ2#0

(343) Rf‘ S17 ’ i™‰j—z[͵Áŧ7€‘Q‡™νg—eπ 5 - GAC GAAGACGGTGAAGAAGG-3

(344) O—lg—eπ5 -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€‘ Q‡™Lʃ0ĭȠ5 PCR L̞.+˱L̵+ PCR ʂɵL

(349) ̇ďTldR‰LıA 1%OX•™e^“¤1ΞȴɁċ#

(350) Ħ̬ď"I+y—rL LAS 500νGE “e]Oξ 5EG̴·#+Ư

(351) Image J Lʃ0y—r7ů΂L̞.+. (13) in situ xQ’kQh™c—. 15.

(352)  in situ xQ’kQh™c—5ʃH UVJ ò˝÷ɳ8

(353) Yoshimura et al.ν2000ξ7 ǰȾ5EGÊǀ#+ MCT4 7O—lg—e€•™ν5 - CGCAAAGGAACACTGG GCTCCCAGCAGCAGACAACCCGTTGGCCA -3 , 5 - ATTGCTTCTGCCAGGAGCA CCAAGCCGATAGCACTG -3 , 5 - AAACGCCGTGCGGGTTTTGTGCCGTGCTCAC ACACA -3 , 5 - TGTGACCAGCTGAGGGCAGTGCCTGCGATGTGCCTTTC -3 ξ8

(354) j™ˆs“pVYc’ u[”Vld“q‘—e~S‘™hνa™‹~PmcŒ™aQ T—oP~Pm[ξ5EG[33P] dATPνNEN ‘Q~aQT—e€•k[n

(355) ‡alŽ™ gmnƒξȡ̈́#+C7LÌʃ#+ 42C 1›ǷxQ’kQh™c—#

(356) Ųɕ1 30 öΎ7ɃɇL 2 Ł̞.+7-

(357) 55˚C 1 40 öΎğƸ"'+ e‘Qr\‘eLɃɇ# +7-

(358) X ˨~P“‰νźŘ~P“‰

(359) ɗđξ5ƽå"'

(360) űÈΫƴΉνM165FCξ1 ̮Ż#+. (14) RNA-Seq ʃ cDNA ‘Q‘’7ȟˌ  RNA-Seq ̰ȍ5ʃHeq‘—rɶʍʔ‘Q‘’7ȟˌ8

(361) SureSelect strand-specific RNA library prep kitνOd”—q˜o[w•d™

(362) W’~U“tOƒξL ʃ

(363) ̼ǴǾ5ư.0̞.+ è RNAν400 ngξF†’νAξͻøLıA mRNA L ŁĠE:Ǯɳď#

(364) O[lw‡Qc— Dνc\‡O“r’ml

(365) ʚλđξ7Ũňď1 Ǯɳď#+ mRNA L͕͠î#+7-¬ȆΈ cDNA Līǀ#+ cDNA 7ȅʿÕƳE :Opt—ºĈL̞.+7-

(366) SureSelect Oligo Adaptor L‘Q^™c—# cDNA 5O k€j™ͻøLºĈ#+ Q—pm[e€‘Q‡™LıA 3 /7€‘Q‡™Lʃ0O k€j™ºĈ cDNA L PCR 5EGŖƛ#

(367) RNA-Seq ̰ȍ5Ìʃ%H cDNA ‘Q‘’ 2#+ MiSeqνQ“ˆs

(368) ɗđξLʃ0 SST E: UVJ ˎ̂ʖʅȋ cDNA ‘Q ‘’7c™]—c—\ν76 bp

(369) ƒOT—rξL̞.+. 16.

(370) (15) RNA-Seq

(371) de novo Og—’E:pP~N”—cŒ“̰ȍ  cutadaptνMartin, 2011ξLʃ

(372) 蒙rͻøFOk€j™ͻø

(373) Æ[V’oPνQV < 30ξ7ȅʿE: 76 ʌʚ7ȅʿLΕĜ#

(374) 50 bp ¼ž7ʣ’™rLʥȚ%H. 25. EG

(375) ’™rL[’™—Om€#+ ͛Ɲ

(376) ȥ¢»c™]—a™Lʃ0Rf‘7r‘ ~q^w‰̰̽"I0HνGenBank ʒ·ʌĨπDRA000595ξνKawahara-Miki et al., 2013ξ ). 1

(377) ùB5Rf‘r‘~q^w‰L’~N”—e2#0Og—’L̞

(378). Tophat E: Cfflinks Lʃ0q‘—e[’€q™‰p™j7pP~N”—cŒ“̰ȍ L̞.+νTrapnell et al., 2012ξ ##4F

(379) ’~N”—e^w‰2#0ʃ+Rf ‘r‘~q^w‰18ʑɽƖLțõ%H+B7’™rēö5‡m}—\14. + Rf‘508

(380) ȥ¢»c™]—a™7DŽ̟2Ȳ͗#0

(381) Trinity 1ʁǀ#+ε[ V’oP7q‘—e[’€q™‰FOg—“#+’™r RNA-Seq ̰ȍ5Ͳ#0 H. 2ǴF54.0HνFinseth and Harrison, 2014ξ #+.0

(382) Trinity 1q. ‘—e[’€q™‰7 de novo Og—’L̞

(383) Og—’#+ͻøL’~N”—e 2#0ʑɽƖ̰ȍL̞.+ %4K-

(384) Trinityνversion: trinityrnaseq-r20140717ξ νGrabherr et al., 2011ξLʃ0[’™—Om€#+’™r7 de novo Og—’L̞

(385) megablast 5EG SILVA rRNA p™j‚™eνQuast et al., 2103ξ2ɭī#

(386) rRNA ʅȋ7ͻøLΕ Ś#+ Trinity 7€‘\Q—Lʃ0 rRNA ͻøLΕĜ#+’™rL

(387) BowtieνLangmead et al., 2009ξLʃ0‡m}—\#+7-

(388) RSEMνLi and Dewey, 2011ξLʃ0͕î ɵ7ŨňȲLǙů#+ SST 2 UVJ ˎ̂ʖ7͵Áŧʑɽ7Ɩ8 edgeR zm]™d νRobinson et al., 2010ξLʃ0ȸů#+ ?+

(389) ̰ː˒E:Æ;˖5Ώͥ%H͵Á ŧgmq7 SST 2 UVJ ˎ̂ʖ5HʑɽƖLʨ̹%H+B

(390) PAGE ̰ȍνparametric analysis of gene set enrichmentξνKim and Volsky, 2005ξL̞.+ q‘—e[’€q™ ‰7 GOνgene ontologyξOwo™c—8

(391) InterproScanνhttp://www.geneontology.org/ external2go/interpro2goξLʃ0̞.+. 17.

(392) (16) ˛ˬÆ;˖7țõ  ͥʂRf‘5 180 mg/kg BW 7 HypoxiprobeTM-1νďīɵĮπpimonidazole HCl

(393) sl Ž‘“˜~N“‡cO˜Q—j™scs“

(394) ‡alŽ™gmnƒξLΠ˽ɀž#

(395) 2 ǶΎƯ5ǮΨƇȮ#0 UVJ LǗġ#+ _—q•™“˱2#0

(396) ʁɿήŔȵLĭ΂Π ˽ɀž#+˱L̵+ Ǘġ#+ UVJ L OCT _—zR—r5ĎŌ#

(397) 80˚C 1ò˝" '+Ư

(398) [’VejmqLʃ0Ě" 10 μm 7ò˝÷ɳLÊǀ#+ ò˝÷ɳLȶðO gq—1Ņů#+7-

(399) ɑº7̼ǴǾ5ư

(400) LJ pimonidazole LJÈLʃ+æʎ˛ˬď ũȒ̍L̞.+ 1 μg/ml 7 DAPI 1ȖȒ̍#+Ư

(401) ̛åΫƴΉνBX51

(402) V’—ze

(403) ǯŷđξLʃ0̮Ż#+. (17) ˆq_—r’OɅƹɖů  SST 7ˆq_—r’OɅƹɖů8

(404) JC-1νďīɵĮπ5,5 ,6,6 -tetrachloro-1,1 ,3,3 -tetraethylbenzimidazolylcarbocyanine iodide

(405) a™‹~PmcŒ™aQT—oP~Pm[

(406) ‡alŽ™gmnƒξLʃ0̞.+ %4K-

(407) ͥʂŠeLǮΨƇȮ#+7- UVJ LǗġ#0lŽ™5ġG

(408) 50 μl 7 HBSS LĈ̰ĂyaˆLʃ0˙÷#+ JC-1 LıA HBSS L 500 μl ĈνȀ˚ɦƢ 125 nMξ

(409) 37˚C 1 20 öΎQ—YŽ‚™q#0̍ ˖L•™pP—\#+7-

(410) 800 × g

(411) Ųɕ1 2 öΎͰƶöΜ#0 UVJ ˛ˬɳLȹɣ" ' 1 ml HBSS 1Ƀɇ#+ _—q•™“˱2#0

(412) ɃɇǶ5ˆq_—r’Oì̂7˾ê Ƭă1H carbonyl cyanide m-chlorophenylhydrazoneνCCCPρc\‡O“r’mlξL 10 μM ɑĈ#+˱L̵+ 800 × g

(413) Ųɕ1 2 öΎͰƶöΜ#+7-

(414) ȹɣ#+ UVJ ˛ˬɳLe‘Qr\‘e5‡R—q#

(415) ̛åΫƴΉνBX51ξLʃ0̮Ż#+ ʈá LΫƴΉʃpdj“WŠ‘νDP70

(416) V’—zeξ5EG̴·#+Ư

(417) Image J Lʃ0. 18.

(418) ̛åƨƢLů΂ď#

(419) 590nm E: 530nm 5H̛åƨƢ7ȲνFI ȲπF590/F530ξL ˈõ#+ FI Ȳ7ˈõ8

(420) SST E: UVJ ʖ˙˺5/0̞.+. (18) ˠ̳̰ȍ  űγ˱Ύ7ȁƼƖ8

(421) öǨöȍȾE: Tukey 7ś΀țůȾ5EGúů#+ ˏŧ7 ͪċOmgQL̞.+űγ5/08

(422) Mann-Whitney 7 U țůȾ5EGˠ̳ʔ4ȁƼ ƖLúů#+. 19.

(423) ˁœˊ ˝ȏ (1). ^“ɧͬ[•‡q\‘~P™5EH UVJ NJõɵ7öΜ.  UVJ NJõɵFͪċDžþłŧLöΜ%H+B

(424) ?&^“ɧͬ[•‡q\‘~P™5E HöʈL̞.+ UVJ NJõɵL Superdex 200pg W‘‰Lʃ+^“ɧͬ[•‡q\‘~ P™5Î%H2

(425) ɛõÈʷ 45 mlν}™[ 1ρöŧ΂ 1,000 kDaξ

(426) 70 mlν}™[ 2 E : 3ρ50 kDaξ

(427) 100-120 mlν}™[ 4 E: 5ρ10 kDa ¼žξº͛5¥̩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+ (p™j8ʪ#04) 7öʈL

(431) HPLC 5EG"F5 öΜ#+. (2). HPLC 5EH UVJ NJõɵ7öΜ.  UVJ NJõɵ7 Superdex 200pg }™[ 4 L

(432) C-22 W‘‰Lʃ+ 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 W‘‰5ȩ3Ôǐ"I4˖͢Göʈ58ȝƹεɵ͍ı?IH2ªƻ"I++ B

(436) Φʝ˒7 PLC 5EHöΜL̸@+. (3). PLC 5EH UVJ NJõɵ7öΜ.  C-22 7ͪċDžþɅƹöʈL PLC 5Î#

(437) c’WLöġ#0NJõ#+Ư

(438) TLC 5E. 0öΜLʨ̹#+ TLC öȍ18’—‹’p—;5EHʑ̍@FI

(439) öʈ 7 E : 8 50ʡĘ1ʪ#+Ė›7e†mqțõ"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) VQ“ɷ7ɌÈ24.+ 7. öʈ5ı?IHɵ͍L

(443) NMR eƒ[q“5EG̰ȍ#+. (4). NMR 5EHˏŧͪċDžþłŧ7ȟͤȸů.  öΜ#+ˏŧͪċDžþöʈ5/0

(444) Īʵ NMR eƒ[q“7ɖůL̞.+ 1H-NMR eƒ[q“E: 1ț˘#+2. 13. C-NMR eƒ[q“7]ˆW“c~qÙLeƒ[q“p™j‚™e. J

(445)

(446) §;E: D-\“_™e7]ˆW“c~qÙ)I*I›. ̉#+νp™j8ʪ#04ξ %4K-

(447) ˏŧͪċDžþöʈ58Z;

(448) §;E: D-\“_™eı?I0+. (5). SST 5H§;7ů΂.  SST 5H§;7Ũň΂L̾<H+B

(449) ”™b™‡Q[•kQg[c—Lʃ0 ÷Gõ#+ SST ʖ5ı?IH§;΂Lɖů#+ UVJ ò˝÷ɳLΫƴΉž1̮Ż% H2 SST E: UVJ ˎ̂ʖ̮Ż"I++BνFig. 2-4, Bξ

(450) SST 7ĵŃ5”™b™ 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 ˎ̂ʖNJõɵ7§;ɦƢν˓ 3 mMξ2Ȳ͗#0 ȁƼ5εÙLʪ#+νFig. 2-4, Aξ. (6). ĖΜ#+ SST 7§;ʂʁ˻.  Ǘġ#+ UVJ L_‘^s™hôɿ#űÈΫƴΉž1̮Ż#+2. J

(454) ĖΜ"I+ SST. ̮Ż"I+νFig. 2-5, Aξ ĖΜ#+ SST L HBSS 5ŁĠ#

(455) 39˚C 1Q—YŽ‚™q #4F§;ɦƢLɖů%H2

(456) ŎΰɌ¤7§;΂˜Ƕʔ5ŖĈ#+νFig. 2-5, Bξ 2-DOG Ɍ¤1 SST LQ—YŽ‚™q#̰ː˒7ͽ˖LΓŴ%H2

(457) §;ɦƢ7dz8. 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.  _‘^s™hôɿ5EGĖΜ#+ 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 7y—r̮Ż"I+νFig. 2-6ξ ͕͠îL̞.04˱ν” ™—π ξ18&ICy—r8@FI4.+  in situ xQ’kQh™c—5EG MCT4 mRNA 7ƃňLʨ̹#+2. J

(464) O—l. g—e€•™Lʃ+˱50c\s“țõ"I+νFig. 2-7, Aξ

(465) g—e€• ™Lʃ+˱18c\s“8țõ"I4.+νFig. 2-7, Bξ űÈΫƴΉž1 X ˨~ P“‰E:e‘QrL̮Ż%H2

(466) SST ʖE: UVJ ˎ̂ʖ5c\s“@F I+νFig. 2-7, C E: Dξ. (8). RNA-Seq Lʃ+ SST ʖ2 UVJ ˎ̂ʖ7͵ÁŧʑɽƖ̰ȍ.  SST ś΂7§;Lʂʁ%HŠWtf‰Lªɖ%H+B

(467) RNA-Seq Lʃ+͵Áŧ ʑɽ7ˤ˯ʔ̰ȍL̞.+ µŁ7űγ1ƱFI+p™jgmq8

(468) DDBJ Sequence Read Archiveνhttp://trace.ddbj.nig.ac.jp/dra/index.htmlξ5ʒ·#+νʒ·ʌĨπDRA003919ξ YiYs™hD„e„~“[qYs™h

(469) }“|—;p{r•^s™hYs™h νPDKξ

(470) §;˾ȵ˖ͽ˖νLDHξ2.+̰ː˒7ͽ˖7ʑɽ΂8

(471) SST 2 UVJ ˎ̂ ʖ27Ύ1ȁƼƖ8@FI&

(472) \“_™eq‘—e†™j™νGLUTξ5/0Cȁ. 22.

(473) ƼƖ8țõ"I4.+νFig. 2-8ξ SST q‘—e[’€q™‰7p™jgmqLʃ 0 KEGG PATHWAY ‡m}—\̰ȍνhttp://www.genome.jp/kegg/pathway.htmlξ

(474) E: SST 2 UVJ ˎ̂ʖ7 GO gmqLʃ+͵ÁŧʑɽƖ̰ȍL̞.+

(475) ̰ː˒7° ͧLʪ%˝ȏ8ƱFI4.+νTable 1ξ ›ǰ

(476) SST 50

(477) Mě͵Áŧ1H c-Fos E: c-Jun 7ʑɽ UVJ ˎ̂ʖ2Ȳ͗#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\s“̮Ż "I+

(480) UVJ ˎ̂ʖ508c\s“̮Ż"I4.+νFig. 2-9, Aξ ?+

(481) ɛť7@Lɀž#+_—q•™“˱18

(482) SST ʖ

(483) UVJ ˎ̂ʖ7&ICc\s“ 8@FI4.+νFig. 2-9, Bξ %4K-

(484) SST ʖ8Æ;˖ɷƾ5I0H. 2. K.+. (10) SST ʖ˙˺7ˆq_—r’OɅƹ  SST ʖ7ˆq_—r’OɅƹL̾<H+B

(485) JC-1 Lʃ0 UVJ LȒ̍#+ SST ʖ8˦̛̍åƨ

(486) ͏̛̍å8=2M3@FI4.+

(487) UVJ ˎ̂ʖ8͏̛̍ åƨ̮Ż"I+νFig. 2-10, Aξ CCCP LɑĈ%H2

(488) SST ʖ

(489) UVJ ˎ̂ʖ2 C5͏̛̍å=2M3@FI44.+νFig. 2-10, Bξ Image J Lʃ0 530 nmν˦ ̍ξE: 590 nmν͏̍ξ5H̛åƨƢLů΂ď#

(490) FI ȲLˈõ#+2. J

(491) SST. ʖ7 FI Ȳ8 UVJ ˎ̂ʖ2Ȳ͗#0ȁƼ5Æ

(492) %4K-ˆq_—r’OɅƹÆ. 23.

(493) . 2ʪ"I+νFig. 2-10, Cξ CCCP LĈ+27 SST ʖE: UVJ ˎ̂ʖ. 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) ຠ  æČćē

(497) Ĕàq½ÉëĎh¢ÐÙÛrÆÉ»fa à&Æ .ÆÉÑʺon´2^ē ĔÐf™Æº€5¡!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.. 

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(500)  ÎÏÜÙ¾cƺ€5Ñ¡!HàÑ\«ÍšÆÉ»rÀÙÓÐon` H›ØÚÝÉ»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. 

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(504) rÀÙÓÑb)j».

(505) A18. B. Lactic acid (mM). 15 12 9. C. 6. *. 3 0 SST s. Epithelial cell.  

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(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.  

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(524) MCT1. MCT2. MCT3. MCT4. − + − + − + − +. S17 M. − +.  £ £

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(528) A. B. C. D D. 

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(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  V“ÐÙÛº‘E Ñ7/ēĔÖÉÒ±7/ēĔÐÀÄܺ

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(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.

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