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Assist. Prof. Yoshiaki Nakayama, Ph. D.
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Ǯǩŀ ŠňŐèrìÂßéƮºżƍÿÎá*Ǟǋŀŉ ł ŀţ Œ Ŝ Ŗ Ş Ń ŀŔ š Ś Ţ Š š ņ Ŕ ť ų ǁ ǃ Ǌ ǜ Ʈ º ż ƍ ǅ ǖ Ǡ Ǖ ŀŀ ŀ Ʋǉǁǟ ŘťŭŞŒŦńţ ǕưǛǡǧL ½  ƙ ü ŀŀŀŀŀǢƙË Ô ŀŀ Î á ß é ƙ ê ƥ! Fig. 1* ƙĂ  Ö Ƙº żƨƫƪǪN*êƮƉƪǚƴǃèrìÂ¼ ŠňŐ ßéƚǪǣǈǐƳŀ wźƁƅƫƠƔƘ,ĲǦ6WƇƍǅǖǠǕƲǉǁǟ
ŘťŭŞŒŦńţ ƱƳǆǀƳǜƙŽƏƔŃŀ ³ƘƌƙêƁƀƐƓżƗż ŘťŭŞŒŦńţ ƘƑżƓŃŀ ƽǐǜã ı | ø ƔŻƪţŒŜŖŞ ¦ƥ ŔšŚŢŠšņŔťų ǁǃǊǜƮºżƓǪƌƙL½-T18 are catalytically inactive, while GalNAc-T9 andüƮćơƍǫƅƫƠƔƘ ŘťŭŞŒŦńţŐ ƥńţňŏ Ƙ[Ɖ-T17, which were identified by us, are brain-specific andƪ ţŒŜŖŞ ǘƻǇǧǪŔšŚŢŠš ǘƻǇǧƮØƇǪǅǖǠǕ

biologically important for the neural differentiation.

Ʋǉǁǟ1 Ú - Ƙ ĝ r ƘÃ ¶ ƆƋƍƕƅƬǪ Ä ġ !

Based on these backgrounds, we have focused on theƘL½Ʈ\ƉƪƅƕƘv%ƇƓżƪǫŀ

functions of glycosyltransferases to make O-glycan

Ŋǩŀ ŠňŐ èrìÂßéƮºżƍÿÎá*Ǟǋŀcarbohydrate-linkage

structures,

andobtained

thefollowing findings.

ŀ ŀ ǢƙËÔŀŀ ŀ ŠňŐ ßéƙmƙ*¦ƚǪǣǈǐƳǥģUB1) Analysis of mice defective of a GalNAc-T17 geneƔ© Ĝ G ĺ ƇǪß é  ı I Ʈi v ƆƋƪ ¦ Ɓº żƨ

We found that GalNAc-T17 KO mice showedƫƓƂƍǫƇƀƇƅƙ¦ƔƚǪĴÎáÜßéƁK£Ɖ

decreased body weight compared with wild-type miceƪƅƕǪv ± ƇƍÎ á ß é Ɲƙ(Fig.

* Ƙĩ  ī ƙG ĺ Ɓ1).

AnalysisofGalNAc-T17expressionpýƔŻƪÖƙ=Ķ¯ƁŻƐƍǫwźƚǪ©ĜGĺƁĴdemonstrated that it was expressed not only inÎ á ß é ƙK £ ƙ. @ ƕæ žǪ È G ĺ ƮH  ƕƇƍ

hippocampus and amygdala, but in ventromedial nucleusÉīƔ(µƙðżÎáßé*ÜƮËÔƇƍǫƅƫ

of hypothalamus, which is known as satiety center.

* ƘĬ ƭƪ@ T ƙÊ Ò ƘH ƒƂǪŁŬłŀGalNAc-T17 may therefore positively regulate feedingƠƔƙÎ á ß éǠǛǏǥƮƾǧǌƇƍG ĺ Å  Ɣ, ` G ĺ ƉƪǪŁŬŬłŀ Ƴǥbehavior.

ǃǡǥƥǌǠǥǃǕƵǡǥÖƙÎá2) Production of zebrafish GalNAc-T mutants*@TƮ8Ƣ°ö¬

To produce zebrafish mutants defective of GalNAc-TGCƮºżƪǪŁŬŬŬłŀ ǣǈǐƳǥģƘ&žƓǪǐǉǈǁƼǎǢĭY$ƔŻƪ ŕŒŠţ Ʈ«&ƉƪǪŁŬŵłŀ*ƙ"¤İƘ ŗŘŗŏ Ʈ«&ƉƪÖƙƮƕƩƫƍǫƌƙâǪ

genes, we employed recently developed genome-editing
technology, TALEN and CRISPR/Cas systems.

We*Ċ\ĪQk Ŋŋ ÆƘÎáÓđƙivǪō Æfound that mutations were successfully introduced at theƘƚƧƩ¨ÃƗÓđgƁāZƆƫƍǫ¿ñƥƸǢtarget sites in the zebrafish embryos using TALEN andǁƴǜƳǝǧǂǥƼƗƖƘƧƪĂƘƧƩǪō ÆƔïj

CRISPR/Cas vectors for GalNAc-T9 and -T18.

ÄƗÎá¨)ƮƇƍv±ÎáßéƝƙ*Ɓĉƣƨ3) A rapid and efficient method for neuronalƫƪƔǪƼǡƱßéƚUBƇƗƀƐƍǫ

ƧƩǪinduction of P19 embryonic carcinoma cell lineƙīƔǪWe revised a novel method to efficiently differentiate*ƉƪƅƕƁƨƀƕƗƐƍǫŀP19 cells into neurons by an adherent culture inƇż¦ƔƚǪŠňŐ ßéƚmƙÝ+(µƧƃÎáßéƘƙơƙşńƼǡƾǁǍEÛĨƙêĂŀlaminin-coated dish in the presence of several reagents toŀ ßéMƔÃ¶Ɖƪÿ şńƼǡƾǁǍEÛĨƙêƮǪ

induce neural differentiation and suppress growth ofǅǖǠǕƲǉǁǟƮºżƓČƞƍǫwźƚƅƙÛĨĚƙ

non-neural cells.

5vƘĬƭƪÛēÐĢÞƁ"èƘſżƓëƘhƃÃwithin 4 days without gliogenesis.

¶ƉƪƅƕƮþżƎƇƍǫƠƍǪĢÞƙÃ¶Ʈ}#Ɖƪ

obtained were responsive to several neurotransmittersƕǪëƘſżƓit½cƮ¸ƈƪƙƘ[ƇǪwith functional neuronal networks.

ǯǩƌƙƙàäƔƙù ¶ E ƙL * ƚā Z ƆƫƗƀƐƍǫƅƙ ƀƨǪƅƙ

We investigated the function of a glycosyltransferase
inResearch projects and annual reports

The mature neurons4) Analysis of extracellular O-glycosylationĢÞƁƘëƘſżƓêƉƪƁÌ<ƆƫƍǫŀŀThe new method induced neurons

zebrafish,

whichO-glycosylation.

catalyzes

extracellularnovel

We found that it was strongly

We have been investigating roles of O-linked sugar

expressed in the brain of zebrafish embryos, and its

chains with the linkage structures, GalNAcα1→Ser(Thr),suppression caused alterations in the brain development.

Manα1→Ser(Thr),

orGlcNAcβ1→Ser(Thr).

O-GalNAc-type sugar chains are called mucin-type96532!

carbohydrates and their biosynthesis is initiated by aY. Nakayama, A. Wada, R. Inoue, K. Terasawa, I. Kimura, N.

group of enzymes, UDP-GalNAc: polypeptide N-acetyl-Nakamura, A. Kurosaka: A Rapid and Efficient Method for

galactosaminyltransferases (GalNAc-Ts).

Neuronal Induction of the P19 Embryonic Carcinoma Cell Line.

GalNAc-Ts

consist of a large gene family with 20 isozymes inhumans.

Interestingly, GalNAc-T8, -T9, -T17, andJournal of Neuroscience Methods, in press.

M. Ogawa, N. Nakamura, Y. Nakayama, A. Kurosaka, H. Manya,

ĿDŀ ǰ¸*VĆč;ǪõVė¾ğR;ŀM. Kanagawa, T. Endo, K. Furukawa, T. Okajima: GTDC2ŀmodifies O-mannosylated ŷ-dystroglycan in the endoplasmic

Ǯǩŀ ƌƙreticulum to generate N-acetyl-glucosamine epitopes reactiveĿDŀ ǰśŢŤ ƿƳƶǥǃƾǧǃǨĠ¹OVĮ_ľ¸lwith CTD110.6 antibody. (2013) Biochem Biophys ResƘ ŘŗŠ ƙÃ¶ƕÚüXĽƮ÷ƐƍǫǩǨŉŇňŊŅŎŅŉōǩǫŀ

Commun 440(1), 88-93ŀŀ

I. Kimura, Y. Nakayama, Y. Zhao, M. Konishi, N. Itoh:
Neurotrophic effects of neudesine in the central nervous system.
(2013) Front Neurosci 7 (article111), 1-5
Y. Nakayama, N. Nakamura, D. Tsuji, K. Itoh, A. Kurosaka:
Genetic diseases associated with protein glycosylation
disorders in mammals. pp. 244-269 in Genetic Disorders,
Maria Puiu (Ed.), ISBN: 978-953-51-08866-3, InTech.
ŀ:632ŀa>, :»ŻƦơ, Ç, ĿDǰ P19ǚƴǃèrì

ÂßéƮºżƍĖęƀƑ(µƙðżÎáĊ\¦ƙËÔ.
Õ36?

T¸²VdǪÎybǪ2013.12.3-6

Y. Nakayama, K. Kato, N. Nakamura, A. Kurosaka: The rolesof a putative polypeptide GalNAc-transferase/Wbscr17.
International Symposium on Glyco-Neuroscience, Awaji city
(Japan) 2014.1.9-11ŀ;614!
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