8/7/2019 A Novel Rat Model with Obesity
1/14
A Novel Rat Model with Obesity-Associated RetinalDegeneration
*HHUHGG\%KDQXSUDNDVK5HGG\ 9LG\XOODWKD9DVLUHGG\ 0G1DZDMHV$0DQGDO 0UXGXOD7LUXYDOOXUX ;LDRIHL):DQJ 0RQLFD0-DEORQVNL *LULGKDUDQ1DSSDQYHHWWLODQG 5DGKD$\\DJDUL
$XWKRU$IILOLDWLRQV
)URPWKH1DWLRQDO,QVWLWXWHRI1XWULWLRQ+\GHUDEDG,QGLD 'HSDUWPHQWRI2SKWKDOPRORJ\DQG9LVXDO6FLHQFHV8QLYHUVLW\RI0LFKLJDQ$QQ$UERU0LFKLJDQDQG 'HSDUWPHQWRI2SKWKDOPRORJ\+DPLOWRQ(\H&HQWHU8QLYHUVLW\RI7HQQHVVHH.QR[YLOOH7HQQHVVHH 3UHVHQWDIILOLDWLRQ'HSDUWPHQWRI2SKWKDOPRORJ\8QLYHUVLW\RI&DOLIRUQLDDW6DQ'LHJR/D-ROOD&DOLIRUQLDDQG
'HSDUWPHQWRI2SKWKDOPRORJ\'HDQ$0F*HH(\H,QVWLWXWH8QLYHUVLW\RI2NODKRPD+HDOWK6FLHQFHV&HQWHU2NODKRPD&LW\2NODKRPD
1H[W6HFWLRQ
Abstract
385326($VWURQJDVVRFLDWLRQEHWZHHQUHWLQDOGHJHQHUDWLRQDQGREHVLW\KDVEHHQVKRZQLQKXPDQV+RZHYHUWKHPROHFXODUEDVLVRILQFUHDVHGULVNIRUUHWLQDOGHJHQHUDWLRQLQREHVLW\LVXQNQRZQ7KXVDQDQLPDOPRGHOZLWKREHVLW\DQGUHWLQDOGHJHQHUDWLRQZRXOGJUHDWO\DLGWKHXQGHUVWDQGLQJRIREHVLW\DVVRFLDWHGUHWLQDOGHJHQHUDWLRQ7KHUHWLQDODEQRUPDOLWLHVLQDQRYHOUDWPRGHO:1,12EZLWKVSRQWDQHRXVO\GHYHORSHGREHVLW\DUHGHVFULEHG
0(7+2'6+LVWRORJLFDQGLPPXQRKLVWRFKHPLFDOH[DPLQDWLRQZHUHSHUIRUPHGRQUHWLQDOVHFWLRQVRIWRPRQWKROG:1,12E UDWV DQG ILQGLQJVZHUHFRPSDUHGZLWKWKRVH RIOHDQ OLWWHUPDWH FRQWUROV 51$ IURPUHWLQDV RIPRQWKROG:1,12EDQGOHDQOLWWHUPDWHUDWVZDVXVHGIRUPLFURDUUD\DQGT573&5DQDO\VLV
5(68/767KH:1,12EUDWVGHYHORSHGVHYHUHREHVLW\ZLWKDQRQVHWDWDSSUR[LPDWHO\GD\V(YDOXDWLRQRIUHWLQDO
PRUSKRORJ\ LQWRPRQWKROG:1,12EDQGDJHPDWFKHGOHDQOLWWHUPDWHFRQWUROVUHYHDOHGSURJUHVVLYHUHWLQDOGHJHQHUDWLRQZLWKDQRQVHWEHWZHHQWRPRQWKVRIDJH ,PPXQRKLVWRFKHPLFDODQDO\VLVZLWKDQWLUKRGRSVLQDQWLFRQHRSVLQDQG36'DQWLERGLHVIXUWKHUFRQILUPHGUHWLQDOGHJHQHUDWLRQSDUWLFXODUO\URGFHOOORVVDQGWKLQQHURXWHUSOH[LIRUPOD\HULQWKHREHVHUDWUHWLQD*HQHH[SUHVVLRQE\PLFURDUUD\DQDO\VLVDQGT573&5HVWDEOLVKHGDFWLYDWLRQRIVWUHVVUHVSRQVHWLVVXHUHPRGHOLQJLPSDLUHGSKRWRWUDQVGXFWLRQDQGSKRWRUHFHSWRUGHJHQHUDWLRQLQ:1,12EUDWUHWLQD
&21&/86,216:1,12EUDWVGHYHORS LQFUHDVHGVWUHVVLQUHWLQDOWLVVXHDQGSURJUHVVLYHUHWLQDOGHJHQHUDWLRQDIWHUWKHRQVHWRIVHYHUHREHVLW\7KH:1,12EUDWLVWKHILUVWUDWPRGHOWRGHYHORSUHWLQDOGHJHQHUDWLRQDIWHUWKHRQVHWRIREHVLW\7KLVQRYHOUDWPRGHOPD\EHDYDOXDEOHWRROIRULQYHVWLJDWLQJUHWLQDOGHJHQHUDWLRQDVVRFLDWHGZLWKREHVLW\LQKXPDQV
3UHYLRXV6HFWLRQ1H[W6HFWLRQ
Introduction
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
8/7/2019 A Novel Rat Model with Obesity
2/14
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
VHYHUDO SRSXODWLRQV REHVLW\ LV RQH RI WKH EDVLF FRPSRQHQWV RI PHWDEROLF V\QGURPH ZKLFK LV LPSOLFDWHG LQPLFURYDVFXODUFKDQJHVLQWKHUHWLQD
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
:LVWDULVWKHROGHVWVWUDLQRIUDWVXVHGLQELRPHGLFDOUHVHDUFK7KH1DWLRQDO,QVWLWXWHRI1XWULWLRQ1,1LQ,QGLDKDVDQ LQEUHG VWRFN RI :LVWDU UDWV GDWLQJ EDFN WR WKDW LV FKULVWHQHG:1,1 :LVWDU PDLQWDLQHG DW 1,1 $
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
9LHZODUJHUYHUVLRQ
y ,QWKLVSDJHy ,QDQHZZLQGRZ
),*85(
L:1,12EUDWLQFRPSDULVRQZLWKLWVOLWWHUPDWHOHDQFRQWURODWWKHDJHRIPRQWKV$:1,12EUDW%/HDQFRQWUROLL5HODWLYHLQFUHDVHLQERG\ZHLJKWVRI:1,12EUDWFRPSDUHGZLWKOHDQFRQWURODQG:LVWDUFRQWUROV9DOXHVUHSUHVHQWPHDQr6'RIDQLPDOVLQHDFKJURXS
3UHYLRXV6HFWLRQ1H[W6HFWLRQ
8/7/2019 A Novel Rat Model with Obesity
3/14
Materials and Methods
Reagents and Antibodies
7KHGHWDLOVRIDQWLERGLHVXVHGIRUWKLVVWXG\DUHDVIROORZVUDEELWDQWLRSVLQUHGJUHHQGLOXWLRQ&KHPLFRQ7HPHFXOD&$PRXVHPRQRFORQDODQWLUKRGRSVLQGLOXWLRQ&KHPLFRQPRXVHSRVWV\QDSWLFGHQVLW\SURWHLQ36'GLOXWLRQ6LJPD$OGULFK6W/RXLV02DQWLUDEELW$OH[D)OXRUGLOXWLRQ,QYLWURJHQ0ROHFXODU3UREHV&DUOVEDG&$DQGDQWLUDEELW$OH[D)OXRUGLOXWLRQ,QYLWURJHQ0ROHFXODU3UREHV
Animals and Tissue Collection
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
Histology and Immunohistochemistry
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
RNA Preparation
5HWLQDVGLVVHFWHGIURPWKHH\HVRI:1 ,12E UDWVDQGDJHPDWFKHGOLWWHUPDWH FRQWUROVZHUHFROOHFWHG LQ UHDJHQW7UL]RO,QYLWURJHQIRU51$LVRODWLRQ51$ZDVLVRODWHGDFFRUGLQJWRWKHPDQXIDFWXUHUVLQVWUXFWLRQV
Global Gene Expression by Microarray Analysis
)LYH PLFURJUDPV RI WRWDO UHWLQDO51$ LVRODWHG IURP WZR SRROHG UHWLQDVZDV XVHG WR V\QWKHVL]H GRXEOHVWUDQGHGF'1$ZLWKWKHXVHRIILUVWDQGVHFRQGVWUDQGF'1$V\QWKHVLVNLWV$II\PHWUL[6DQWD&ODUD&$DFFRUGLQJWRWKHPDQXIDFWXUHUVJXLGHOLQHV3XULILFDWLRQRIGVF'1$ODEHOLQJK\EULGL]DWLRQZDVKLQJDQGVWDLQLQJZHUHFDUULHGRXWDFFRUGLQJWRWKH$II\PHWUL[SURWRFRO7KHUDWJHQHH[SUHVVLRQFKLS$II\PHWUL[ZDVXVHGIRUK\EULGL]DWLRQ7KUHHLQGHSHQGHQWVDPSOHVIRUHDFK:1,12EDQGFRQWUROJURXSZHUHDQDO\]HG
*HQH FKLSV ZHUH VFDQQHG 0LFURDUUD\ 6XLWH >$II\PHWUL[@ RSHUDWHG E\ *&26 VRIWZDUH YHUVLRQ DQGH[SUHVVLRQGDWDZHUHQRUPDOL]HGZLWKWKHXVHRIUREXVWPXOWLDUUD\DYHUDJH'DWDZHUHEDVHGRQWKUHHJHQHFKLSVSHU HDFK FRQWURO DQG :1,12E JURXSV 5DWLRV RI DYHUDJH VLJQDO LQWHQVLW\ ORJWUDQVIRUPHG GDWD ZHUH WKHQFDOFXODWHGIRUWKHSUREHVHWV:1,12EUHODWLYHWROHDQFRQWURODQGZHUHFRQYHUWHGWRIROGFKDQJH7KHFRQWUDVWEHWZHHQH[SHULPHQWDODQGFRQWUROZDVH[WUDFWHGVHOHFWLQJWKRVHJHQHVZ LWKDQDGMXVWHG3RIDQGDWKUHHIROGGLIIHUHQFH
Quantitative Real-Time PCR
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
8/7/2019 A Novel Rat Model with Obesity
4/14
&W EHWZHHQ FRQWURO DQG :1,12E JURXSV DIWHU QRUPDOL]DWLRQ WR YDULRXV KRXVHNHHSLQJ JHQHVK\SR[DQWKLQHSKRVSKRULERV\OWUDQVIHUDVH+JSUWDFWLQVXFFLQDWHGHK\GURJHQDVH6GK$DQGULERVRPDOSURWHLQ/5SODVGHVFULEHG0HDQr6'YDOXHVRISHUFHQWDJHH[SUHVV LRQRYHUFRQWUROOHDQFDOFXODWHGIURPDWOHDVWWKUHHLQGHSHQGHQWVDPSOHVDUHSUHVHQWHG
3UHYLRXV6HFWLRQ1H[W6HFWLRQ
Results
Morphologic Evaluation
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
9LHZODUJHUYHUVLRQ
y ,QWKLVSDJHy ,QDQHZZLQGRZ
),*85(
5HSUHVHQWDWLYHKLVWRORJ\RIFRQWUROUDWUHWLQD$&(DQG:1,12EUDWUHWLQD%')DWYDULRXVDJHV$PRQWKROGFRQWURO%PRQWKROG:1,12E&PRQWKROGFRQWURO'PRQWKROG:1,12E(PRQWKROGFRQWURO)PRQWKROG:1,12E+LVWRJUDPVVXPPDUL]LQJWKHSURJUHVVLRQRIUHWLQDOGHJHQHUDWLRQLQFHQWUDOLLDQGSHULSKHUDOLLLUHWLQDRI:1,12EUDWVFRPSDUHGZLWKOHDQFRQWUROV9DOXHVUHSUHVHQWPHDQr6'RIDWOHDVWVHYHQLQGHSHQGHQWREVHUYDWLRQV26RXWHUVHJPHQWV,6LQQHUVHJPHQWV,1/LQQHUQXFOHDUOD\HU,3/LQQHUSOH[LIRUPOD\HU*&JDQJOLRQFHOOOD\HU
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
8/7/2019 A Novel Rat Model with Obesity
5/14
,QDGGLWLRQWRDQDO\]LQJ:1,12EUDWVDQGOHDQOLWWHUPDWHFRQWUROVZHHYDOXDWHGWKHUHWLQDOKLVWRORJ\RIWRPRQWKROG:1,1SDUHQWVWUDLQ UDWV 7KHVH UDWV GLGQRWVKRZVLJQLILFDQW FKDQJHV LQUHWLQDOPRUSKRORJ\GDWDQRWVKRZQVXJJHVWLQJWKDWWKHUHWLQDOGHJHQHUDWLRQREVHUYHG LQWKH:1,12EUDWLVQHLWKHUVSRUDGLFQRUDQRQVSHFLILFREVHUYDWLRQLQWKLVVWUDLQRIUDWV
Immunohistochemistry
,PPXQRKLVWRFKHPLFDODQDO\VLVRI:1,12EDQGOHDQOLWWHUPDWHFRQWUROUDW UHWLQDVZDV FDUULHGRXWXVLQJURGDQGFRQH SKRWRUHFHSWRU VSHFLILF PDUNHU DQWLERGLHV DQG DQWL36' DQWLERGLHV ,PPXQRVWDLQLQJ ZLWK UKRGRSVLQDQWLERGLHVVKRZHGUHGXFWLRQLQWKHEULJKWQHVVRIWKHUKRGRSVLQVSHFLILFVLJQDOLQWKH:1,12EUDWUHWLQDFRPSDUHGZLWKWKDWRIOHDQFRQWUROVDWPRQWKVGDWDQRWVKRZQDQGPRQWKV)LJV$%&',PPXQRVWDLQLQJZLWKFRQHRSVLQ0:DQWLERGLHV LQGLFDWHGQRVLJQLILFDQWDOWHUDWLRQ LQWKHQXPEHURIFRQHV LQWKH:1,12EUDWUHWLQD)LJV()*+ VWDLQLQJWKHUHWLQDOVHFWLRQVZLWKVKRUWHUZDYHOHQJWKFRQHRSVLQ6:JDYHVLPLODUUHVXOWVGDWDQRWVKRZQ7KHQXPEHURIFRQHVLQWKHUHWLQDVRI:1,12EDQGOHDQFRQWUROUDWVZDVIRXQGWREHVLPLODUDWDJHVWRPRQWKV)LJLL
9LHZODUJHUYHUVLRQ
y ,QWKLVSDJHy ,QDQHZZLQGRZ
),*85(
,PPXQRKLVWRFKHPLFDOHYDOXDWLRQRIURGDQGFRQHSKRWRUHFHSWRUPDUNHUVLQ:1,12EUDWFRPSDUHGZLWKFRQWUROVDWPRQWKVL5HWLQDOVHFWLRQVRI$&(*FRQWURODQG%')+:1,12EUDWVZHUHODEHOHGZLWK$'UKRGRSVLQDQG(+FRQHRSVLQ1XFOHLDUHODEHOHGZLWK'$3,6FDOHEDUPLL+LVWRJUDPGHPRQVWUDWHVWKHQXPEHURIFRQHVLQ:1,12EDQGFRQWUROUDWUHWLQDV9DOXHVUHSUHVHQWPHDQr6'RIWKUHHLQGHSHQGHQWREVHUYDWLRQV3
7RIXUWKHUHYDOXDWHWKHGHFUHDVHREVHUYHGKLVWRORJLFDOO\LQWKHWKLFNQHVVRI23/ZHODEHOHGWKHUHWLQDOVHFWLRQVIURP:1,12EDQGFRQWUROUDWVZLWKSRVWV\QDSWLFGHQVLW\SURWHLQ36'DQGFRPSDUHGWKH LPPXQRIOXRUHVFHQFH)LJ,QWKHPRQWKROG:1,12EUDWUHWLQDWKH LPPXQRIOXRUHVFHQFHRI36'ZDVZHDNHU)LJV&'WKDQWKH36'VLJQDOREVHUYHGLQDJHPDWFKHGOHDQOLWWHUPDWHV)LJV$% 7KLV LQGLFDWHGDORVVRIV\QDSWLFWHUPLQDOVDQGIXUWKHUVXSSRUWHGWKHREVHUYHGUHGXFWLRQLQWKHWKLFNQHVVRI23/LQ:1,12EUDWUHWLQD
8/7/2019 A Novel Rat Model with Obesity
6/14
9LHZODUJHUYHUVLRQ
y ,QWKLVSDJHy ,QDQHZZLQGRZ
),*85(
,PPXQRIOXRUHVFHQFHLPDJHVRIPRQWKROGFRQWURO$%DQG:1,12EUDW&'UHWLQDOVHFWLRQVODEHOHGZLWK36'/DEHOLQJRI36'LVSURPLQHQWDQGLQWHQVHLQ23/OD\HURIFRQWUROVZKHUHDVLQWKH:1,12EUDWWKHIOXRUHVFHQFHVLJQDORI36'LVOHVVLQWHQVH
Differential Expression of Genes in WNIN-Ob Retina
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
PRQWKROG:1,12E UDWV )LJ 2EHVLW\DVVRFLDWHG JHQHV WXEE\ DQG %EVDOVRVKRZHG D ORZHU OHYHORIH[SUHVVLRQ LQWKHUHWLQDVRI:1,12EUDWVWKDQLQWKRVHRIOLWWHUPDWH FRQWUROV )LJEXWWKHGHFUHDVHZDVQRWVLJQLILFDQW
8/7/2019 A Novel Rat Model with Obesity
7/14
9LHZODUJHUYHUVLRQ
y ,QWKLVSDJHy ,QDQHZZLQGRZ
),*85(
4XDQWLWDWLYHH[SUHVVLRQRIURGVSHFLILFJHQHVLQPRQWKROG:1,12EUDWVDQGWKHLUOHDQOLWWHUPDWHFRQWUROV([SUHVVLRQYDOXHVZHUHGHWHUPLQHGE\T573&5DQGSUHVHQWHGRQDQDUELWUDU\VFDOH\D[LVDIWHUQRUPDOL]DWLRQZLWKUDW+JSUW'DWDUHSUHVHQWWKHPHDQr6'RQDQDUELWUDU\VFDOHDQGZHUHFDOFXODWHGIURPDWOHDVWWKUHHLQGHSHQGHQWREVHUYDWLRQV
9LHZODUJHUYHUVLRQ
y ,QWKLVSDJHy ,QDQHZZLQGRZ
),*85(
([SUHVVLRQRI&U[(ORYODQGREHVLW\UHODWHGJHQHVLQWKHUHWLQDVRIPRQWKROG:1,12EUDWVDQGWKHLUOHDQOLWWHUPDWHFRQWUROV([SUHVVLRQYDOXHVZHUHGHWHUPLQHGE\T573&5DQGSUHVHQWHGRQDQDUELWUDU\VFDOH\D[LVDIWHUQRUPDOL]DWLRQZLWKUDW+JSUW'DWDUHSUHVHQWWKHPHDQr6'RQDQDUELWUDU\VFDOHDQGZHUHFDOFXODWHGIURPDWOHDVWWKUHHLQGHSHQGHQWREVHUYDWLRQV
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
8/7/2019 A Novel Rat Model with Obesity
8/14
9LHZODUJHUYHUVLRQ
y ,QWKLVSDJHy ,QDQHZZLQGRZ
),*85(
4XDQWLWDWLYHH[SUHVVLRQRIFRQHVSHFLILFJHQHVLQPRQWKROG:1,12EUDWVDQGWKHLUOHDQOLWWHUPDWHFRQWUROV([SUHVVLRQYDOXHVZHUHGHWHUPLQHGE\T573&5DQGSUHVHQWHGRQDQDUELWUDU\VFDOH\D[LVDIWHUQRUPDOL]DWLRQZLWKUDW+JSUW'DWDUHSUHVHQWWKHPHDQr6'RQDQDUELWUDU\VFDOHDQGZHUHFDOFXODWHGIURPDWOHDVWWKUHHLQGHSHQGHQWREVHUYDWLRQV
9LHZODUJHUYHUVLRQ
y ,QWKLVSDJHy ,QDQHZZLQGRZ
),*85(
4XDQWLWDWLYHH[SUHVVLRQRIVHOHFWHGVWUHVVUHODWHGJHQHVLQUHWLQDVRIPRQWKROG:1,12EUDWVDQGWKHLUOHDQOLWWHUPDWHFRQWUROV([SUHVVLRQYDOXHVZHUHGHWHUPLQHGE\T573&5DQGSUHVHQWHGRQDUELWUDU\VFDOH\D[LVDIWHUQRUPDOL]DWLRQZLWKUDW+JSUW'DWDUHSUHVHQWWKHPHDQr6'RQDQDUELWUDU\VFDOHDQGZHUHFDOFXODWHGIURPDWOHDVWWKUHHLQGHSHQGHQWREVHUYDWLRQV
3UHYLRXV6HFWLRQ1H[W6HFWLRQ
Discussion
5HWLQDOGHJHQHUDWLRQLQFOXGLQJDJHUHODWHGPDFXODUGHJHQHUDWLRQ$0'DQGGLDEHWLFUHWLQRSDWK\'5 LVDPDMRUFDXVHRI EOLQGQHVV LQ WKH GHYHORSHGDQG WKH GHYHORSLQJZRUOG 7KH PROHFXODU HYHQWV WKDW FRQWULEXWH WR UHWLQDOGHJHQHUDWLRQ DUH KLJKO\ FRPSOH[ EHFDXVH RI WKH LQYROYHPHQW RI GLIIHUHQW UHWLQDO FHOO W\SHV 0XWDWLRQV LQDSSUR[LPDWHO\ JHQHV KDYH EHHQ LPSOLFDWHG LQ UHWLQDO GHJHQHUDWLRQ 5HW1HWZZZVSKXWKWPFHGX5HWQHWKRPHKWP'HVSLWHH[WHQVLYHNQRZOHGJHDERXWWKHGLVHDVHFDXVLQJJHQHVWKHPROHFXODUEDVLVXQGHUO\LQJPDQ\IRUPVRIUHWLQDOGHJHQHUDWLRQDUHQRWXQGHUVWRRG7KHLQWHUSOD\EHWZHHQPXOWLSOH
8/7/2019 A Novel Rat Model with Obesity
9/14
JHQHWLFDQGHQYLURQPHQWDOIDFWRUVFRXOGDGGWRWKHFRPSOH[LW\IRUWKHSDWKRJHQHVLVRIUHWLQDOG\VWURSKLHV$VWURQJDVVRFLDWLRQIRXQGEHWZHHQUHW LQDOGHJHQHUDWLRQDQGREHVLW\IXUWKHUDGGVWRWK LVFRPSOH[LW\7KHDODUPLQJLQFUHDVHLQWKHSUHYDOHQFHRIREHVLW\LQ6RXWKHDVW$VLDQFRXQWULHVIXUWKHUH[DFHUEDWHVFRQFHUQDERXWUHWLQDOGHJHQHUDWLRQJOREDOO\7KHUHIRUHLGHQWLI\LQJDVXLWDEOHDQLPDOPRGHOZLOODLG LQ LQYHVWLJDWLQJWKHXQGHUO\ LQJPHFKDQLVPIRUWKHDVVRFLDWLRQEHWZHHQUHWLQDOGHJHQHUDWLRQREHVLW\DQGPHWDEROLFV\QGURPH
(YDOXDWLRQRIWKHPRUSKRORJ\SKRWRUHFHSWRUVSHFLILFSURWHLQDQGJHQHH[SUHVV LRQSURILOHRIWKH:1,12EUDWUHWLQD
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
5HWLQDODEQRUPDOLWLHVZHUHREVHUYHG LQDIHZ URGHQWPRGHOVZLWKREHVLW\7KHWXEE\PLFHGHYHORSQHXURVHQVRU\DEQRUPDOLWLHV DQG REHVLW\EXW QHXURVHQVRU\ DEQRUPDOLWLHV GHYHORSPXFK HDUOLHUWKDQWKH RQVHW RIREHVLW\ 7KHREHVLW\RIWXEE\PLFHLVUHODWLYHO\PLOGKDVODWHRQVHWDQGSURJUHVVHVVORZO\ 0RXVHPRGHOVIRU%DUGHW%LHGOV\QGURPH GHYHORS REHVLW\ DQG UHWLQDO DEQRUPDOLWLHV LQ DGGLWLRQ WR D VSHFWUXP RI RWKHU SKHQRW\SLF IHDWXUHV $PRQJRWKHU REHVHURGHQWPRGHOV RFXODU FRPSOLFDWLRQV SDUWLFXODUO\ SURQRXQFHGUHWLQDOFKDQJHV ZHUH IRXQG LQ
GLDEHWLF REHVH IDID UDWVZKHQ WKH\ZHUH IHG DVXFURVH GLHWIRUZHHNV0LFHODFNLQJ IXQFWLRQDO WXEE\OLNHSURWHLQ 78/3 GHYHORSUHWLQDO GHJHQHUDWLRQEXWQRWREHVLW\ 7KH SKHQRW\SH REVHUYHG LQ:1,12E UDWV LVXQLTXHEHFDXVHRIWKHHDUO\DJHRIRQVHWDQGWKHVHYHULW\RIREHVLW\DQGUHWLQDOGHJHQHUDWLRQ
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
7KH XSUHJXODWHG JHQHVDUHPRVWO\UHODWHG WRDSRSWRWLF VWUHVVUHVSRQVH WLVVXH UHPRGHOLQJSDWKZD\V DQQH[LQFDWKHSVLQJOXWDWKLRQH6WUDQVIHUDVH&S*IDSLQGLFDWLQJDFWLYDWLRQRIWKHVWUHVVUHVSRQVHLQWKH:1,12EUDWUHWLQD
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
8/7/2019 A Novel Rat Model with Obesity
10/14
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
2EUDWPD\DLGLQYHVWLJDWRUVWRDGGUHVVWKHVHLVVXHV
3UHYLRXV6HFWLRQ1H[W6HFWLRQ3UHYLRXV6HFWLRQ1H[W6HFWLRQ
Acknowledgments
7KHDXWKRUVWKDQN%RLQGOD6HVLNHUDQ1DWLRQDO,QVWLWXWHRI1XWULWLRQIRUDVVLVWDQFHLQH[DPLQLQJWKHKLVWRORJ\VOLGHV.ULVKQDNXPDU 6DUDGD DQG 3DVXSXOHWL /RSDPXGUD 1DWLRQDO ,QVWLWXWH RI 1XWULWLRQ IRU DVVLVWDQFH LQ SUHSDULQJKLVWRORJLF VSHFLPHQV0RWKD 6DW\DYDQL 1DWLRQDO ,QVWLWXWH RI 1XWULWLRQ IRU SURYLGLQJ DQLPDO PDLQWHQDQFH DQGWHFKQLFDODVVLVWDQFH$XVWUD/LHSD8QLYHUVLW\RI0LFKLJDQIRUDVVLVWDQFH LQPDQXVFULSWSUHSDUDWLRQDQG0LWFKHOO*LOOHW8QLYHUVLW\RI0LFKLJDQIRUKLVWRORJLFWLVVXHSUHSDUDWLRQ
3UHYLRXV6HFWLRQ1H[W6HFWLRQ
Footnotesy 7KHVH DXWKRUV FRQWULEXWHG HTXDOO\ WR WKH ZRUN SUHVHQWHG KHUH DQG VKRXOG WKHUHIRUH EH UHJDUGHG DVHTXLYDOHQWDXWKRUV
y 6XSSRUWHGLQSDUWE\WKH,QGLDQ&RXQFLORI0HGLFDO5HVHDUFKDQG'HSDUWPHQWRI%LRWHFKQRORJ\1HZ'HOKL1DWLRQDO ,QVWLWXWHV RI+HDOWK *UDQWV(
8/7/2019 A Novel Rat Model with Obesity
11/14
0HGOLQH $OOLVRQ'%)RQWDLQH.50DQVRQ-(6WHYHQV-9DQ,WDOOLH7%$QQXDOGHDWKVDWWULEXWDEOHWRREHVLW\LQWKH8QLWHG6WDWHV-$0$$EVWUDFW)5(()XOO7H[W
0RNGDG$+0DUNV-66WURXS')*HUEHUGLQJ-/$FWXDOFDXVHVRIGHDWKLQWKH8QLWHG6WDWHV
-$0$$EVWUDFW)5(()XOO7H[W 2JGHQ&/&DUUROO0'&XUWLQ/50F'RZHOO0$7DEDN&-)OHJDO.03UHYDOHQFHRIRYHUZHLJKWDQGREHVLW\LQWKH8QLWHG6WDWHV-$0$$EVWUDFW)5(()XOO7H[W
9LOODUHDO '7$SRYLDQ &0 .XVKQHU 5) .OHLQ 6 2EHVLW\ LQ ROGHU DGXOWV WHFKQLFDO UHYLHZ DQGSRVLWLRQVWDWHPHQWRI WKH$PHULFDQ6RFLHW\ IRU1XWULWLRQDQG1$$627KH2EHVLW\6RFLHW\2EHV5HV0HGOLQH:HERI6FLHQFH
+RVVDLQ 3 .DZDU % (O 1DKDV 0 2EHVLW\ DQG GLDEHWHV LQ WKH GHYHORSLQJ ZRUOGD JURZLQJFKDOOHQJH1(QJO-0HG&URVV5HI0HGOLQH:HERI6FLHQFH
3UHQWLFH $0 7KH HPHUJLQJ HSLGHPLF RI REHVLW\ LQ GHYHORSLQJ FRXQWULHV ,QW - ( SLGHPLRO$EVWUDFW)5(()XOO7H[W
&KHXQJ1:RQJ7
8/7/2019 A Novel Rat Model with Obesity
12/14
.OHLQ53HWR7%LUG$9DQQHZNLUN057KHHSLGHPLRORJ\RIDJHUHODWHGPDFXODUGHJHQHUDWLRQ$P-2SKWKDOPRO&URVV5HI0HGOLQH:HERI6FLHQFH
.ULVKQDLDK6'DV71LUPDODQ3.HWDO5LVNIDFWRUVIRUDJHUHODWHGPDFXODUGHJHQHUDWLRQILQGLQJVIURP WKH $QGKUD 3UDGHVK (\H 'LVHDVH 6WXG\ LQ 6RXWK ,QGLD ,QYHVW 2SKWKDOPRO 9LV 6FL
$EVWUDFW)5(()XOO7H[W
%ODFTXH 2( /HURX[ 05 %DUGHW%LHGO V\QGURPH DQ HPHUJLQJ SDWKRPHFKDQLVP RI LQWUDFHOOXODUWUDQVSRUW&HOO0RO/LIH6FL&URVV5HI0HGOLQH:HERI6FLHQFH
+DJVWURP 6$ 1RUWK 0$ 1LVKLQD 3/ %HUVRQ (/ 'U\MD 73 5HFHVVLYH PXWDWLRQV LQ WKH JHQHHQFRGLQJWKHWXEE\OLNHSURWHLQ78/3LQSDWLHQWVZLWKUHWLQLWLVSLJPHQWRVD1DW*HQHW&URVV5HI0HGOLQH:HERI6FLHQFH
2KOHPLOOHU..+XJKHV50/HWW-0HWD O3URJUHVVLRQRIFRFKOHDUDQG UHWLQDOGHJHQHUDWLRQ LQ WKHWXEE\UGPRXVH$XGLRO1HXURRWRO0HGOLQH
*LULGKDUDQ 19$QLPDO PRGHOV RI REHVLW\ DQG WKHLU XVHIXOQHVV LQ PROHFXODU DSSURDFK WR REHVLW\,QGLDQ-0HG5HV0HGOLQH:HERI6FLHQFH
-D\DUDPDQ.66XPRUDWVVHWUHVHDUFKHUVRQKXQWIRUREHVLW\JHQHV1DW0HG0HGOLQH
*LUGKDUDQ19+DULVKDQNDU16DW\DYDQL0$QHZUDWPRGHOIRUWKHVWXG\RIREHVLW\6FDQG-/DE$QLP6FL
0DQGDO 01$PEDVXGKDQ5:RQJ 3:*DJH 3-6LHYLQJ 3$$\\DJDUL5&KDUDFWHUL]DWLRQ RIPRXVH RUWKRORJXH RI (/29/ JHQRPLF RUJDQL]DWLRQ DQG VSDWLDO DQG WHPSRUDO H[SUHVVLRQ*HQRPLFV&URVV5HI0HGOLQH:HERI6FLHQFH
0DQGDO019DVLUHGG\9-DEORQVNL00HW DO6SDWLDO DQG WHPSRUDOH[SUHVVLRQRI0)53 DQG LWVLQWHUDFWLRQZLWK&753,QYHVW2SKWKDOPRO9LV6FL$EVWUDFW)5(()XOO7H[W
0DQGDO019DVLUHGG\95HGG\*%HWDO&753LVDPHPEUDQHDVVRFLDWHGDQGVHFUHWRU\SURWHLQLQ WKH 53( DQG FLOLDU\ ERG\ DQG WKH 65 PXWDWLRQ RI &753 LPSDLUV LWV VHFUHWLRQ ,QYHVW2SKWKDOPRO9LV6FL$EVWUDFW)5(()XOO7H[W
9DVLUHGG\ 9 -DEORQVNL 00 0DQGDO 01 HW DO (ORYO ESGHOHWLRQ NQRFNLQ PLFH GHYHORSSURJUHVVLYHSKRWRUHFHSWRUGHJHQHUDWLRQ,QYHVW2SKWKDOPRO9LV6FL$EVWUDFW)5(()XOO7H[W
8/7/2019 A Novel Rat Model with Obesity
13/14
&URVV5HI0HGOLQH:HERI6FLHQFH 5DWWQHU $ 6XQ + 1DWKDQV - 0ROHFXODU JHQHWLFV RI KXPDQ UHWLQDO GLVHDVH $QQX 5HY *HQHW&URVV5HI0HGOLQH:HERI6FLHQFH
5LYROWD&6KDURQ''H$QJHOLV00'U\MD735HWLQLWLVSLJPHQWRVDDQGDOOLHGGLVHDVHVQXPHURXV
GLVHDVHVJHQHVDQGLQKHULWDQFHSDWWHUQV+XP0RO*HQHW$EVWUDFW)5(()XOO7H[W
0LVUD$3DQGH\ 50'HYL -56KDUPD5 9LNUDP1. .KDQQD1 +LJK SUHYDOHQFH RI GLDEHWHVREHVLW\DQGG\VOLSLGDHPLDLQXUEDQVOXPSRSXODWLRQLQQRUWKHUQ,QGLD,QW-2EHV5HODW0HWDE'LVRUG&URVV5HI0HGOLQH:HERI6FLHQFH
8/7/2019 A Novel Rat Model with Obesity
14/14
9D]TXH]&KRQD)6RQJ%.*HLVHUW((-U7HPSRUDOFKDQJHVLQJHQHH[SUHVVLRQDIWHULQMXU\LQWKHUDWUHWLQD,QYHVW2SKWKDOPRO9LV6FL$EVWUDFW)5(()XOO7H[W
.LP/6)LVKPDQ*$6HLSOH:+6]O\N-36WRQH(05HWLQDOG\VIXQFWLRQ LQFDUULHUVRI%DUGHW%LHGOV\QGURPH2SKWKDOPLF*HQHW&URVV5HI0HGOLQH:HERI6FLHQFH
+DPPRQG%5-U&LXOOD7$6QRGGHUO\'00DFXODUSLJPHQWGHQVLW\LVUHGXFHGLQREHVHVXEMHFWV,QYHVW2SKWKDOPRO9LV6FL