• Ei tuloksia

DENDRITIC SPINE MORPHOGENESIS

52/(2)1(8521$/,J6)$'+(6,2102/(&8/(6$1'10'$5(&(37256,17+(&16

2. DENDRITIC SPINE MORPHOGENESIS

2.1. STRUCTURE OF DENDRITIC SPINES

'HQGULWLFVSLQHVDUHGHULYHGIURP¿ORSRGLDWKHLPPDWXUHIRUPRIGHQGULWLFVSLQH 7KH\ DUH YHU\ VPDOO SURWUXVLRQV PP ZKLFK DUH H[SUHVVHG RQ QHXURQDO GHQGULWHVLQWKHEUDLQ7KH\FRQWDLQWKHSRVWV\QDSWLFFRPSRQHQWZKLFKLVLPSRUWDQW IRUH[FLWDWRU\V\QDSVHVLQWKHEUDLQ+DUULVDQG.DWHU+RZHYHUSRVWV\QDSWLF PDFKLQHU\LVODFNLQJIURPWKHLPPDWXUHGHQGULWLF¿ORSRGLD7KHVSLQHVDUHNQRZQ WREHLQYROYHGLQV\QDSWLFIRUPDWLRQDQGSODVWLFLW\'HQGULWLFVSLQHVFRQWDLQFRPSOH[

VWUXFWXUHVZLWKGLIIHUHQWWUDQVPHPEUDQHF\WRVNHOHWDODQGVFDIIROGLQJPROHFXOHV 7KHDPRXQWRI¿ORSRGLDDQGVSLQHVYDULHVGHSHQGLQJRQKRZPXFKLQIRUPDWLRQ LVSDVVLQJWKURXJKWKHV\QDSVHV8VXDOO\VSLQHVGRPLQDWHRYHU¿ORSRGLDZKHQ V\QDSWLFWUDQVPLVVLRQWDNHVSODFH'HQGULWLFVSLQHDEQRUPDOLWLHVDUHIRXQGLQPDQ\

QHXURORJLFDODQGSV\FKLDWULFGLVHDVHV&DODEUHVHHWDO

'HQGULWLFVSLQHVFDQEHGLYLGHGLQWRWKUHHGLIIHUHQWW\SHVWKHLPPDWXUHWKLQ W\SHWKHVWXEE\W\SHDQGWKHPDWXUHPXVKURRPW\SH)LJ$7KHWKLQW\SHLV FKDUDFWHUL]HGE\DVPDOOKHDGDQGDVOHQGHUQHFNZKHUHDVWKHPXVKURRPW\SHKDV DODUJHKHDGDQGDVKRUWQHFNZLWKWKHVWXEE\W\SHEHLQJDQLQWHUPHGLDWHIRUP ODFNLQJWKHQHFN6HNLQRHWDO'HSHQGLQJRQWKHVLWXDWLRQDQ\W\SHRIWKHVH VSLQHVFDQGRPLQDWHRYHUWKHRWKHUV,QWHUPHGLDWH¿ODPHQWVDQGPLFURWXEXOHVDUH QRWIRXQGLQVSLQHVEXWWKH\FDQEHIRXQGLQODUJHQXPEHUVLQWKHGHQGULWLFVKDIW +XVLHWDO

7KHDFWLQF\WRVNHOHWRQDQGDVWUXFWXUHFDOOHGSRVWV\QDSWLFGHQVLW\36'DUH WZRPDMRUVWUXFWXUDOFRPSRQHQWVRIGHQGULWLFVSLQHV)LJ%DQGLQQHXURQV DFWLQLVH[SUHVVHGDEXQGDQWO\)LODPHQWRXVDFWLQ)DFWLQLVDEOHWRIRUPGLIIHUHQW QHWZRUNVWKDWDUHWKHPDLQFRPSRQHQWVRIWKHDFWLQF\WRVNHOHWRQ&DODEUHVHHWDO 6HNLQRHWDO0RUSKRORJLFDOFKDQJHVLQVSLQHVWUXFWXUHVDUHPRGXODWHG E\DFWLYDWHGJOXWDPDWHUHFHSWRUV0DWXV

36' FDQ EH IRXQG XQGHU WKH SRVWV\QDSWLF PHPEUDQH RI GHQGULWLF VSLQHV .HQQHG\9DULRXVVFDIIROGSURWHLQVDQGVLJQDOOLQJPROHFXOHVFDQEHIRXQG LQWKH36'6KHQJVXFKDVVHYHUDO36''OJ$]R3'=SURWHLQVDQG PDQ\PHPEUDQHDVVRFLDWHGJXDQ\ODWHNLQDVHV0$*8.V.LPHWDO36' ZKLFKLVDW\SLFDO3'=SURWHLQLVDEXQGDQWO\H[SUHVVHGLQWKH36'&KRHWDO .LVWQHUHWDO

$IWHUVSLQHPRUSKRJHQHVLVKDVEHJXQWKHUHLVV\QDSWLFDVVHPEO\RI36' 2NDEHHWDO6WXGLHVRQ36'NQRFNRXWPLFHLQGLFDWHWKDWWKH\H[KLELW QRDEQRUPDOLWLHVLQVSLQHVWUXFWXUHV0LJDXGHWDO,QFRQWUDVWWKHDFWLQ F\WRVNHOHWRQLVLQYROYHGLQFRQWUROOLQJVSLQHPRUSKRORJ\DVWKHGLVUXSWLRQRIWKHDFWLQ

F\WRVNHOHWRQFDQLQWHUIHUHZLWKVSLQHPRUSKRJHQHVLVDQGGLVDVVHPEO\RIV\QDSWLF HOHPHQWV$OOLVRQHWDO=KDQJDQG%HQVRQ7KHUHIRUH36'PD\KDYH DUROHLQVSLQHPRUSKRJHQHVLVEXWWKHDFWLQF\WRVNHOHWRQKDVDPRUHGRPLQDQWUROH

2.2. DENDRITIC SPINE DEVELOPMENT

$WWKHEHJLQQLQJRIV\QDSWRJHQHVLVGHQGULWLFVKDIWVFRQWDLQQXPHURXV¿ORSRGLD 7KH\DUHORQJDQGWKLQSURWUXVLRQVDQGWUDQVLHQWLQWKHLUQDWXUHDQGWKH\FRQWDLQ OHVV)DFWLQWKDQVSLQHV)LORSRGLDSUHVHQWLQJURZWKFRQHVDUHXVXDOO\LQYROYHG LQD[RQDODQGGHQGULWLFRXWJURZWK)LORSRGLDLQGHQGULWLFVKDIWVSDUWLFLSDWHLQ V\QDSWRJHQHVLV3RUWHUD&DLOOLDXHWDO

Fig 6. Structure of dendritic filopodia and spines, and a model of an asymmetric synapse. (A) Filopodia are the precursors of dendritic spines, and they develop into three types of dendritic spines: thin type, stubby type and mushroom type spines. Gray disks: PSD structure and red circles: F-actin.

(B) The synapse is formed between the presynaptic terminal and the dendritic spine. Synaptic vesicles are found in the presynaptic terminal. Two structural elements: actin cytoskeleton and PSD are important for the function of the dendritic spine. PSD contains the neurotransmitter receptors.

Actin-binding proteins, including drebrin, are stabilizing the dendritic spine architecture. Modified from Sekino et al, 2007.

'(1'5,7,&63,1(0253+2*(1(6,6

'XULQJV\QDSVHIRUPDWLRQWKHVSLQHVVWDUWWRGRPLQDWHRYHU¿ORSRGLD0DWXV 8VXDOO\ZKHQ¿ORSRGLDDUHFRQYHUWHGWRVWDEOHVSLQHVWKHUHPD\EHDWUDQVLWLRQ VWDJHLQYROYHG'DLOH\DQG6PLWK'XQDHYVN\HWDO7ZRW\SHVRI

¿ORSRGLDFDQEHIRXQGFOXVWHUW\SHDQGGLIIXVHW\SH7DNDKDVKLHWDO7KH FODVVL¿FDWLRQGHSHQGVRQKRZGUHEULQELQGLQJ)DFWLQ'%)DFWLQLVFOXVWHUHG 'UHEULQLVXVXDOO\KLJKO\H[SUHVVHGLQWKHEUDLQ+D\DVKLHWDODQGLWFRQWUROV VSLQHPRUSKRJHQHVLV0L]XLHWDO7DNDKDVKLHWDO

'LIIXVHW\SH¿ORSRGLDFDQEHIRXQGLQWKHHDUO\SRVWQDWDOEUDLQDQGWKHDPRXQWLV KLJKDWWKLVVWDJH'UHEULQ(LVH[SUHVVHG¿UVWWKLVEHLQJIROORZHGE\GUHEULQ$$RNL HWDO:KHQGUHEULQIRUPVFOXVWHUVZLWK)DFWLQWKHFOXVWHUW\SH¿ORSRGLD GHYHORS36'FOXVWHUVIRUPQH[WZLWKFRQFRPLWDQWIRUPDWLRQRIVSLQHV'UHEULQ LVDOVRWUDQVORFDWHGIURPVSLQHVWRGHQGULWHVZKHQ10'$5VEHFRPHDFWLYDWHG 6HNLQRHWDO

$W WKH EHJLQQLQJ RI V\QDSWRJHQHVLV ¿ORSRGLD DUH H[SUHVVHG RQ QRQVSLQ\

QHXURQV:RQJDQG:RQJ$VWKHQHXURQDOPDWXUDWLRQFRQWLQXHVWKH QXPEHURIFOXVWHUW\SH¿ORSRGLDLQFUHDVHVDQGWKH\KDYHPRUHSUHV\QDSWLFFRQWDFW VLWHV7DNDKDVKLHWDO$VWKHFOXVWHUW\SH¿ORSRGLDODFN36'FOXVWHUV WKH\DUHEHOLHYHGWREHWKHWUDQVLWLRQDO¿ORSRGLDW\SHEHIRUHWXUQLQJLQWRVSLQHV 6HNLQRHWDO1RQHWKHOHVVRQO\VRPHSRUWLRQRI¿ORSRGLDDUHWXUQHGLQWR VSLQHV=LYDQG6PLWK7KHUHIRUHPDQ\IDFWRUVVXFKDV&$0VDQGVLJQDOOLQJ PROHFXOHVDUHWKRXJKWWREHLQYROYHGLQFRQWUROOLQJV\QDSWRJHQHVLVDWGLIIHUHQW GHYHORSPHQWDOVWDJHV

7KHDPRXQWRIVSLQHVLVKLJKHVWLQWKHODWHUGHYHORSPHQWDOVWDJHVZKHQV\QDSWLF SODVWLFLW\LVDOVRKLJK'HQGULWLFVSLQHIRUPDWLRQFRQWLQXHVDWLWVPD[LPXPOHYHO XQWLOWKHWKLUGSRVWQDWDOZHHNLQURGHQWKLSSRFDPSXV7KHVSLQHGHQVLW\UHPDLQV DWDVWDEOHOHYHOLQDGXOWKRRG=KDQJDQG%HQVRQ+RZHYHUPXOWLSKRWRQ PLFURVFRSLF VWXGLHV KDYH VKRZQ WKDW VSLQH DQG V\QDSVH IRUPDWLRQ FRQWLQXHV WKURXJKRXWOLIH7UDFKWHQEHUJHWDO7KHUHIRUHWKHDGXOWEUDLQLVDEOHWR IRUPQHZV\QDSWLFFRQQHFWLRQVDOOWKHWLPH

2.3. REGULATION OF DENDRITIC SPINE DEVELOPMENT

7KHIRUPDWLRQRIFRQWDFWVEHWZHHQFHOOVDWV\QDSVHVUHVHPEOHVWKHFRQWDFWIRUPDWLRQ EHWZHHQQRQQHXURQDOFHOOV+RZHYHUWKHV\QDSWLFFRQWDFWVDUHGLIIHUHQWEHFDXVH ERWK WKH SUH DQG SRVWV\QDSWLF HOHPHQWV KDYH SRODUL]HG DV\PPHWULF IHDWXUHV 6HYHUDODGKHVLRQPROHFXOHVVXFKDV1&$0V1FDGKHULQVDQGSURWRFDGKHULQVDUH LQYROYHGLQFRQWUROOLQJV\QDSVHIRUPDWLRQ7DNHLFKLDQG$EH:DVKERXUQHHW DO&DODEUHVHHWDO2WKHUPROHFXOHVDUHDOVRLQYROYHGVXFKDVQHXUH[LQV QHXUROLJLQV(SKUHFHSWRUVDQGWKHLUHSKULQOLJDQGV&DODEUHVHHWDO

&DGKHULQVEHORQJWRWKHIDPLO\RIFDOFLXPGHSHQGHQWKRPRW\SLF&$0VDQG WKH\DUHH[SUHVVHGE\DOPRVWDOOFHOOV)RUH[DPSOH1FDGKHULQLVIRXQGLQQHXURQV 7DNHLFKL DQG $EH :KHQ V\QDSVH IRUPDWLRQ EHJLQV 1FDGKHULQV DUH ORFDWHGRQSUHDQGSRVWV\QDSWLFVLGHVLQWKHQHZO\IRUPHGV\QDSVHPDNLQJVWDEOH FRQQHFWLRQV$FWLQELQGLQJSURWHLQVVXFKDVDDFWLQLQDQGFR¿OLQOLQNWKHFDGKHULQ EFDWHQLQFRPSOH[LQGLUHFWO\WRWKHDFWLQF\WRVNHOHWRQ1HXKRIIHWDO,WKDV EHHQVKRZQWKDWEFDWHQLQKDVDQLPSRUWDQWUROHLQ1FDGKHULQPHGLDWHGDGKHVLRQ GXULQJV\QDSVHIRUPDWLRQ0XUDVHHWDOG&DWHQLQVDUHDOVRELQGLQJSDUWQHUV IRUFDGKHULQVDQGWKH\DUHLQYROYHGLQVSLQHUHJXODWLRQ.RVLNHWDO

7KHSURWRFDGKHULQVEHORQJWRDVXEJURXSRIWKHFDGKHULQVXSHUIDPLO\DQGD DQGJSURWRFDGKHULQVDUHDEXQGDQWO\H[SUHVVHGLQQHXURQVDQGWKH\FDQEHIRXQG LQV\QDSVHV3KLOOLSVHWDO6WXGLHVRQJSURWRFDGKHULQNQRFNRXWPLFHKDYH UHYHDOHGWKDWWKHDPRXQWRIV\QDSVHVLVGLPLQLVKHGLQWKHVSLQDOFRUGVRIWKHVH DQLPDOVLPSO\LQJWKDWFDGKHULQVDUHUHTXLUHGIRUV\QDSWLFGHYHORSPHQW:HLQHU HWDO

6HYHUDO&$0VRIWKH,J6)ZRUNWRJHWKHUWRMRLQFHOOVQHDUWRHDFKRWKHUWKURXJK WKHLUH[WUDFHOOXODUGRPDLQV&$0VLQFOXGLQJ1&$06\Q&$0DQGQHFWLQDUH LQYROYHGLQUHJXODWLQJDGKHVLRQLQWKHV\QDSVHWRJHWKHUZLWKLQWHJULQVWKXVEHLQJ DEOHWRKDYHDQHIIHFWRQGHQGULWLFVSLQHGHYHORSPHQWDQGPRUSKRORJ\%HQVRQHW DO&KDYLVDQG:HVWEURRN:DVKERXUQHHWDO

1HXUH[LQVDQGQHXUROLJLQVDUHLQYROYHGLQVSLQHDQGV\QDSVHIRUPDWLRQDV ZHOO 6FKHLIIHOH 1HXUH[LQV DUH XVXDOO\ ORFDWHG RQ WKH SUHV\QDSWLF VLGH DQGWKH\SURPRWHSRVWV\QDSWLFGLIIHUHQWLDWLRQZKHUHDVQHXUROLJLQVUHVLGHRQWKH SRVWV\QDSWLFVLGHDQGDUHLQYROYHGLQSUHV\QDSWLFGLIIHUHQWLDWLRQ7KH\IXQFWLRQDW ERWKDŽDPLQREXW\ULFDFLG*$%$HUJLFLQKLELWRU\DQGJOXWDPDWHUJLFH[FLWDWRU\

V\QDSVHV*UDIHWDO&DODEUHVHHWDO7KH36'DVVRFLDWLRQZLWK QHXUROLJLQVPD\UHJXODWHQHXUROLJLQGLVWULEXWLRQDQGIXQFWLRQDODFWLYLW\&DODEUHVH HWDO

6HYHUDOH[WUDFHOOXODUO\VHFUHWHGPROHFXOHVLQFOXGLQJEUDLQGHULYHGQHXURWURSKLF IDFWRU%'1)DQGWKURPERVSRQGLQVSDUWLFLSDWHLQV\QDSWRJHQHVLVWRR-LHWDO

&KULVWRSKHUVRQHWDO&DODEUHVHHWDO$PHPEUDQHSURWHLQ V\QGHFDQLVDOVRLQYROYHG(WKHOODQG<DPDJXFKL6WUXFWXUHVUHVHPEOLQJ VSLQHVFDQDOVREHIRXQGLQQHXURQVZKLFKQRUPDOO\ODFNVSLQHVDQGWKHVHDUHWKRXJKW WREHPDGHXSRILQWUDFHOOXODUHOHPHQWV)LQDOO\ELQGLQJRIHSKULQVWRWKHLUW\URVLQH NLQDVHUHFHSWRUV(SKVDOVRPRGXODWHVVSLQHPRUSKRORJ\E\DFWLYDWLQJYDULRXV VLJQDOOLQJSDWKZD\VLQYROYLQJWKH5KRIDPLO\*73DVHV&DODEUHVHHWDO

'(1'5,7,&63,1(0253+2*(1(6,6

2.4. REGULATION OF DENDRITIC SPINE MOTILITY AND STABILIZATION

7KHDFWLQF\WRVNHOHWRQLVLQYROYHGLQUHJXODWLQJVSLQHPRWLOLW\VWDELOLW\DQGVKDSH

&LQJRODQLDQG*RGD&DGKHULQVQHXUROLJLQVDQG(SKUHFHSWRUVLQGLUHFWO\

OLQN WKH DFWLQ F\WRVNHOHWRQ WR WKH QHXURWUDQVPLWWHU UHFHSWRUV 'UHEULQ FR¿OLQ SUR¿OLQJDQGJHOVROLQDUHLQYROYHGLQFRQWUROOLQJDFWLQSRO\PHUL]DWLRQ*OXWDPDWH UHFHSWRUVLQFOXGLQJ10'$5VSDUWLFLSDWHLQVSLQHRXWJURZWKUHJXODWLRQ0DWXV 7KH&D0.VLJQDOOLQJSDWKZD\FDQEHDFWLYDWHGE\10'$5VDQGWKLVOHDGV WRF\WRVNHOHWDOFKDQJHVEXWLWPD\DOVROHDGWRWKHIRUPDWLRQRIGHQGULWLFVSLQHVDQG V\QDSVHV7KXVWKH&D0.SDWKZD\LVLQYROYHGLQFRQWUROOLQJVSLQHPRUSKRJHQHVLV

&LDQLDQG6DOLQDV

5KRIDPLO\*73DVHVEHORQJWRWKH5DVVXSHUIDPLO\RIVPDOO*73DVHV7KH\

UHJXODWHWKHG\QDPLFVRIWKHDFWLQF\WRVNHOHWRQ+DOODQG1REHV7KH5KR

*73DVHV5DF5KR$DQG&GFDUHNQRZQWRLQÀXHQFHGHQGULWLFVSLQHIRUPDWLRQ DQGVKDSH1HZH\HWDO6WXGLHVRQQHXURQVH[SUHVVLQJFRQVWLWXWLYHO\DFWLYH 5DFVKRZWKDWWKHVHQHXURQVIRUPVPDOOVSLQHV/XRHWDOZKHUHDVLQ QHXURQVH[SUHVVLQJFRQVWLWXWLYHO\DFWLYH5KR$WKHVSLQHGHQVLW\DQGOHQJWKLV GLPLQLVKHG7DVKLURHWDO7KHIXQFWLRQRI&GFLQVSLQHPRUSKRJHQHVLV LVQRWNQRZQ6WXGLHVKDYHVKRZQWKDWFRQVWLWXWLYHO\DFWLYHRUGRPLQDQWQHJDWLYH IRUPVRI&GFGRQRWFKDQJHVSLQHGHQVLW\RUOHQJWK7DVKLURHWDO2QWKH RWKHUKDQGDQRWKHUVWXG\GHPRQVWUDWHGWKDW&GFPD\EHLQYROYHGLQGHQGULWLF VSLQHGHYHORSPHQW,ULHDQG<DPDJXFKL

7KH 5KR *73DVHV PD\ FRQWURO WKH G\QDPLFV RI WKH DFWLQ F\WRVNHOHWRQ E\

UHJXODWLQJVLJQDOOLQJSKRVSKROLSLGVLQFOXGLQJSKRVSKDWLG\OLQRVLWROELVSKRVSKDWH 3,3ZKLFKLVDEOHWRLQGXFHDFWLQSRO\PHUL]DWLRQ:HHUQLQNHWDO7KH SURWHLQP\ULVWR\ODWHGDODQLQHULFK&NLQDVHVXEVWUDWH0$5&.6ELQGV3,3LQD SKRVSKRU\ODWLRQGHSHQGHQWZD\DQGWKXVLWKDVDQLPSRUWDQWUROHLQPDLQWDLQLQJ WKHGHQGULWLFVSLQHPRUSKRORJ\&DODEUHVHDQG+DOSDLQ0$5&.6LVDOVR DEOHWRELQGWRDFWLQ¿ODPHQWVDQG&DFDOPRGXOLQ7KHUHIRUHLWLVFRQVLGHUHG WRPHGLDWH3.&DQGFDOPRGXOLQGHSHQGHQWVSLQHIXQFWLRQV0RUHRYHUWZRRWKHU

*73DVHV5DVDQG5DSSDUWLFLSDWHLQUHJXODWLQJWKHGHQGULWLFVSLQHPRUSKRORJ\

&DODEUHVHHWDO

2.5. DENDRITIC SPINE PATHOLOGY IN NEUROLOGICAL DISEASES

6SLQHGLVUXSWLRQLVDW\SLFDOIHDWXUHIRXQGLQPDQ\SV\FKLDWULFDQGQHXURORJLFDO GLVRUGHUV%ODQSLHGDQG(KOHUV&DODEUHVHHWDOLQFOXGLQJVWURNH HSLOHSV\WUDXPDGHPHQWLDPDMRUGHSUHVVLRQQRUPDODJLQJVFKL]RSKUHQLDDQG

DGGLFWLRQ)LDODHWDO*ODQW]DQG/HZLV1LPFKLQVN\HWDO6ZDQQ HWDO6SLQHPDOIRUPDWLRQVXVXDOO\DIIHFWV\QDSWLFIXQFWLRQV

&KDQJHVLQVSLQHVWUXFWXUHDQGV\QDSWLFSODVWLFLW\PD\EHUHODWHGWRWKHFRJQLWLYH GH¿FLWVREVHUYHGLQGLIIHUHQWQHXURORJLFDOGLVHDVHV,QVFKL]RSKUHQLDWKHDPRXQW RIVSLQHVLVVLJQL¿FDQWO\UHGXFHGLQSUHIURQWDOFRUWLFDOS\UDPLGDOQHXURQV*ODQW]

DQG/HZLV7KHVDPHSKHQRPHQRQLVVHHQLQKXPDQHSLOHSV\6ZDQQHW DODQGLVPLPLFNHGLQDQLPDOPRGHOVRIHSLOHSV\,VRNDZD-LDQJ HWDO,Q$'DQG3DUNLQVRQ¶VGLVHDVHZKLFKDUHFKURQLFQHXURGHJHQHUDWLYH GLVHDVHVWKHQXPEHURIVSLQHVLVUHGXFHGRURWKHUFKDQJHVFDQEHIRXQGLQGHQGULWHV 6HONRH'DWDXVLQJDQ$'DQLPDOPRGHOVKRZHGWKDWZKHQSRVWV\QDSWLF DFWLQUHJXODWRU\PDFKLQHU\LVGLVUXSWHGV\QDSWLFIXQFWLRQVDUHGLVWXUEHGWKLVEHLQJ UHÀHFWHGLQFRJQLWLYHGH¿FLWV

7KHPDMRUKHUHGLWDU\PHQWDOUHWDUGDWLRQIRUPVIUDJLOH;V\QGURPHDQG'RZQ V\QGURPHDUHFKDUDFWHUL]HGE\FKDQJHVLQVSLQHPRUSKRORJ\7KHDPRXQWRIPDWXUH VSLQHVLVGHFUHDVHGDQGWKHUHLVDJUHDWHUQXPEHURI¿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

'(1'5,7,&63,1(0253+2*(1(6,6