筑波大学技術報告24: 5-9, 2004ॶ౗ఝƳൟരơǓ३‫ډ‬ஞૄ൝ࡐƶ୩໷இӕ৖඲ƶӬದ
2๟൉Г௼1¹࢖ߤҥન ¹অ௞ஞў

3૿ౡપԙॶյ੒‫ݜ‬௖ؐА‫ࠊףڽ‬ѬࡋáЌԙ‫ڈ‬âĞ305-8575 Иश‫ۀ‬ƬƗƸࠅஔ҉઩ 1-1-1ਭॎҠԙ৾ॐ඲¹೪௾ЂஇƲ࠺ҫ‫܎ڏ‬ƶऎ‫׫‬඲¹൵ƻƳ௻ǑǔƧൟ৖‫ڤ‬үƳƬƌƮඦ‫ݥ‬ơǓºӶ຤ࠕ‫؁‬Ʒ¹ࣹ௫൝ƶ౗ఝ३‫ډ‬ஞૄ൝ࡐƶൟരǚਙ໹ЏƳ஬ǒ¹ެභȧȔȦƯ୩໷இƳൟ৖ơǓධ඲ǚӬದƟƮƕƧ[1-8]º‫ܮ‬Ƴ¹ƛƶධ඲ǚ֏Ƴॶ౗ǚൟ৖ơǓƧǉƶ Human Brain Mapping Analyzer ǚӬದƟ[9]¹ບÐƲ३‫ډ‬ஞૄ൝ࡐǌƥƶշ༫൝ࡐƶൟരǚӕ৖ƟƮƕƧ [9-16] º‫ݶ‬Ӗ¹ƥƶଇƓǑॶ౗ǚ୩໷ൟ৖ơǓƧǉƶ‫ڸ‬ઑƶ໊ࣕධ඲¹഍෭ƶ߁ৄ඲¹࠺ҫ‫܎ڏ‬ƶࣩ‫׫‬඲‫ח‬ƻ௻ǑǔƧൟ৖‫ڤ‬үƳƬƌƮඦ‫ݥ‬ơǓºŌ¼ƷƠǉƳ३‫ڈډ‬ƶऻඦஞૄƷ¹ǲȅȒǴƳƒƙǓ३‫ډ‬ஞૄ൝ࡐƶࢇࢄƳǐƫƮ݉ƲǗǔƮƌǓºࢫƫƮ¹౗ఝƶԇ൉ϳƳƒƙǓ३‫ډ‬ஞૄ൝ࡐƶൟരǚൟ৖ơǓࠧƷ¹ଇঞౖ֞ǚӕฯơǓरƯ¹ƝǑƳࡾÐƶࡏഗƶದऔ֞ࣥǚӕฯơǓरƯ‫ئ‬ǉƮࢲ຤ƯƊǓºࢫີƶ‫ףڽ‬Ʒ¹ࡶƳিҠԙஇධ඲ưืћਭॎҠԙஇධ඲ƳǐƫƮ݉ƲǗǔƮƕƧºিҠԙஇධ඲Ʒ൝ࡐ໷ǌԩ঴૮ǚ୩໷ơǓƶƳ๲ǔƮƌǓƔ¹ࠟ໬‫ࡷޡ‬ưਕ໊ࣕ੅߁Ưƶ‫ތ܄‬ǚƷƠǉ¹ൟ৖Օஶǌ൝ࡐƶࠑ‫ۼ‬඄঴ƳǐǓ‫ތ܄‬௖ƳǐƫƮ¹়ԏƯधެƲ୩໷૮ǚ௻ǓƛưƔ‫ݷ‬బƯƊƫƧºГධ¹ืћਭॎҠԙஇධ඲ƯƷ¹ԩ঴൝ࡐƶ‫ܨ‬ઑƳઓơǓืћ঴Ʒࠑ‫ۼ‬ೝԔஇସƌ࠯յϩ୩ƟƮƒǒ¹ԇ൝ࡐƶൟരǚެභȧȔȦƯधެƳ¹ƟƓǊ়ԏƳࡀൿơǓƛưƔҧౖƯƊǓºƟƓƟ¹ƥǔǑǚ୩໷இƳൟ৖ơǓƶƳ໴ƌධ඲ƔƲƓƫƧºȁȧȎǦȜȤǚ߹ພƟƧӋ੡ӕ৖ਗ਼૷Ư
Ʒ¹ଽਅ঴¹‫۔ޙ‬঴¹ՕஶƲƱƶ஝ƳƒƌƮ๓ભƔ
ƊǓºǆƧ¹ȧÓǱÓǮȮȐǤÓǦȦ‫ۉ‬೰‫؛‬ƷșǪȨ഍෭ƶൟ৖ƳƷ߹ພƯƕƲƌºࠕ‫؁‬Ʒ¹ࣹ௫൝ƶ౗഍෭ǚ‫ื܎ڏ‬ћਭॎҠԙஇƳ৾ॐƟ¹ƥƶ‫܎ڏ‬؊ஶƶൟരǚެභȧȔȦƯ୩໷ơǓ Brain Mapping Analyzer ǚӬದƟ[1]¹ࡾÐƶ౗ఝ३‫ډ‬Ҡԙ൝ࡐƶൟരǚൟ৖ƟƮƕƧ[1-8]ºƛƶൟ৖඲Ʒ¹ȁȧȎǦȜȤǚພƌƧӋ੡ӕ৖඲ǌ஢֡Ҡԙ‫֎ࢽڸ‬ǚພƌƧ‫ੱݗ‬њઑǪȨșȃǫȤȐǞÓ඲ưೝǁƮ¹ࣺƲƗưǊ )‫ڞ‬ϱर೰ެƯ়ԏƲൟ৖‫ڤ‬үǚ௻ǓƛưƔƯƕǓºƝǑƳ¹ƛƶਗ਼૷ǚ֏Ƴ¹ॶ౗ఝƳൟരơǓ३‫ډ‬Ҡԙ൝ࡐǚൟ৖ơǓƧǉƶ Human Brain
Mapping Analyzer ưƥƶӕ৖඲ǚӬದƟƧ [9] ºƛƶਗ਼૷ƳǐƫƮ¹ॶ౗ƶৡඅਙ฻ǚঝȚǪȨȮ෹Ƴঝഉญ໹ЏƳൟԥƟƮບÐƲ३‫ډ‬Ҡԙ൝ࡐƶൟരǚൟ৖Ɵ¹୩໷இƳӋ੡ҠƯƕǓઁ¹ԇ໹Џƶ‫܎ڏ‬؊ஶ૮ǚଋࢽƟƮ௚‫໊ࣕڐ‬ơǓƛưƔƯƕǓº෭ඦ‫ݥ‬ƯƷ¹ƛƶਗ਼૷ǚ໅ພƟƮॶ౗ǚ୩໷ൟ৖ơǓƧǉƶ‫ڸ‬ઑƶ໊ࣕධ඲¹഍෭ƶ߁ৄධ඲¹ืћō¼഍෭ƶ߁ৄ඲෭ࡑ‫ۊ‬ƯƷ¹३‫ډ‬ഗ༝‫ח‬ƻু३ഗ༝ǚ࠮ƧƲƌ 3ઑƶ૬঴á50-70 ‫ޣ‬âƶ౗ǚ߹ພƟƧºƛǔǑƷƌƢǔǊ૿ౡપԙƳЌԙؐА½‫ףڽ‬ƶƧǉƳЋੵǐǒ‫ڼ‬ઑưƟƮ୲‫׼‬ƝǔƧǊƶƯƊǓº୩໷ൟ৖ǚ݉ƎƧǉƳƷ¹ਭॎǚެ൉ǆƯ‫ج‬ГƳՏਙƳ۞୩ƟƲƙǔƸƲǑƲƌºࣹ௫൝ǚພƌƧࡑ‫ۊ‬ƯƷ¹ਙॱƳ۞୩њǚᓕ໙Ɵ¹ƥƶ‫ۼ‬౗ǚࡷǒࢽƟƮƌƧºƟƓƟ¹ॶƶਙॱǚ۞୩ơǓƳƷ۞୩њƔપ໷Ƴഁ຤ƯƊǓर¹ࠑ‫࠯ۼ‬յƔ‫ډ‬ӄƟƮƌǓƧǉƳે‫ڥ‬¹۞୩ƔƎǆƗƌƓƲƌҧౖ঴ƔƊǓࠧƲƱƓǑ¹౗ǚࡷǒࢽƟƧ‫ۼ‬Ƴ଎హ۞୩ǚ݉ƲƫƧºืћಶ҃঴ƳƷࠑ‫ۼ‬࠯յǊշ‫ٱ‬ƟƮƗǓƛưƓǑ¹ࠑ‫ۼ‬œ࠯յϱఝƶ౗ǚພƌƧº2.1ॶ౗ƶ۞୩౗ƷƯƕǓۙǒ‫ڥ‬ըǚ࣪ƬƙƲƌǐƎܺ෽ǚപƙƧीઙƯࡷǒࢽƟƧºȥȮ߱ժढি໊॒ѻ঎ (PBS,pH 7.4) ƯฏƧƟƧȋǾȃƳǧÓǷǚତǒ¹ƥƶरƳ¹౗ǚໍඉƟƮ૷ƌƧº໩੩ƶఝ‫ڔ‬௫ฝ‫ח‬ƻ౗୫௫ฝƳǦȆȠÓȧǚ੃హƟ¹ȕȥǴǺȘȮȒǚພƌƮ PBSá'͇âǚ෹ൟ๢ 65 ml ƶੱஶƯ଎హƟƮે‫ڥ‬ƟƧº੶ƌƮ )%ȌȤȖȦțǝȦȂȍȄư %#'%ǫȦ
ǺȦǝȦȂȍȄǚսǈ PBS ຜњá6͇âƳǐƫƮ଎హ۞୩ǚ݉ƲƫƧº౗ƶܺ෽ǚƷƢƟ¹ਕԜ૨ƶ‫ܓ‬
Ɲ 1cm ƶȑȨǾǪƳൟԥƟ¹4%ȌȤȖȦțǝȦȂȍȄƶ PBS ຜњá4ąâƯ 2 షյ¹‫۞ۼ‬୩ǚ݉ƲƫƧºƝǑƳ౗ȑȨǾǪǚ PBS ຜњá4ąâƯ 3 షյ৽िƟ¹5%ǲȢ௙ PBS ຜњá4ąâƳ 4 షյ।୑ƟƧºǲȢ௙њǚ।௤ƝƣǓƛưƳǐƫƮ¹஽‫࠯ڤ‬ƳਭॎఝƶഎƶದিǚຫরƟ¹ਭॎƶ੾࣪ǚේƘƛưƔƯƕǓºԇ౗ȑȨǾǪƶఫ෽ư‫ڥ‬ըǚଛౄƳࡷǒࣩƕ¹ȄȤǟǝǟǴरƯ‫ੱכ‬Ƴ஽‫ڤ‬Ɲƣ¹ՄੌǚේƘƧǉธൊƟƮ-80ąƯඐ੻ƟƧº2.2഍෭ಏৡ
෭‫ףڽ‬ƯƷਕԜ૨ȑȨǾǪǚ‫ދ‬Ыವ‫ע‬ƳವৡƟ߹ພƟƧº౗ȑȨǾǪǚӕ஽Ɵ 13%ȘȥȎȆȦǝȦǮ
ÓȦǚսǈ PBS ຜњƳඤ෵Ɵ¹‫ޙ‬ƻ஽‫ڤ‬ƟƧº੶ƌƮ¹પ‫ٸ‬ƶ஽‫ڤ‬ৡඅພȚǪȨȃÓț (Cryo-Microtom,PMV, Sweden) ǚພƌƮ 20 µm ƶ‫ܓ‬ƝƯ༫੶இƳಏৡƟƧºಏৡƟƧ഍෭ǚ PBS ຜњरƳഷƓǁƮƟǗǚॖƸƟ¹঎ຜ঴ƶȘȥȎȆȦǝȦǮÓȦǚࣩ‫׫‬Ɵ¹

1E-mail: yabek@md.tsukuba.ac.jp; Tel: 029-853-3231
E-mail: akiyamak@md.tsukuba.ac.jp; Tel: 029-853-3330
3
E-mail: dsutoo@md.tsukuba.ac.jp; Tel: 029-853-3113
2-5-ǴȤǟȄǧȤǴ (75 x 150 mm) ƳତǒപƙƧºǴȤǟȄǧȤǴƳƷ഍෭ƶƷƔǔǚේࠐơǓƧǉ¹ຊǉ
ǷȤǼȮ"ǪȨțฯᙟຜњáǷȤǼȮ 0.5%,ǪȨțฯᙟ 0.1%âƯǮÓȁǞȮǫǚࠍƟƧºŎ¼೪௾Ђஇ࠺ҫ‫܎ڏ‬ƶऎ‫׫‬඲‫ݗ‬༛ƶॶ౗¹௾ƳಋࡐƶȚǣȥȮఝƳƷ¹ࡲ༛ƶȤǾȃǌșǡǴƳƷ‫ۂ‬ǑǔƲƌ¹ȥȘȐǴǼȮ௖Ƴ຀ີơǓ೪௾ЂஇƲ࠺ҫ‫܎ڏ‬Ɣ੻޲ơǓºƥƶƧǉ¹ࢫີƶ‫ื܎ڏ‬ћਭॎҠԙஇƲൟ৖඲ƯƷॶƶ౗ǚଽৣൟ৖ơǓƛưƷ‫ݷ‬బƯƊƫƧºƥƛƯ¹ॶ౗ƶ࠺ҫ‫܎ڏ‬ǚࣩ‫׫‬ơǓƧǉƳ֕ƬƓƶධ඲ǚࠟǇƧº3.1Ҡԙಶ҃ƳǐǓऎ‫׫‬඲(1) ๺֞ຜ౷ƳǐǓຜӕ¾ǣǺȉÓȦ¹ȜǺȉÓȦฅƷǪȨȨȖȦțƳ഍෭ǚ 30 ൟյØ24 ࠯յ।ƟƧº(2) ߱¹ǝȦǦȥƳǐǓຄ໐½ຜӕ¾1N ѻ߱Ƴ
10 ൟյ¹ϦƌƷ 0.1N ঎߱ҠȅȃȥǡțຜњƳ 5 ൟյ।ƟƧº(3) ߱Ҡ½ճ‫ ¾໊ࣕی‬i) 0.25% ӄșȮǧȮ߱Ǧȥǡț (KMnO4) Ƴ 20 ൟյ।ƟƧ‫ۼ‬¹ृໜ঎Ư৽िƟ¹Pal ຜњá1%ψໝ߱Ǧȥǡțư 1%ǲȠǡ߱ƶ‫ݜށ‬њâƳ 1Ø2 ൟյ।Ɵ¹PBS Ư৽िƟƧºii) ӄ߱Ҡ঎ਬ঎Ƴ 2Ø3 ൟյ।ƟƧºiii) ঎ਬҠǝȦȚȆǡțȥǼǡțຜњฅƷ঎ਬҠȖǡਬȅȃȥǡțຜњƳ 10 ൟյ।ƟƧº(4) ߱Ҡഏಋಶ҃¾ȂǲǬÓǺÓఝƯ౗഍෭ƶ঎ൟǚࢪൟƳࣩ‫׫‬Ɵ¹ශཋѻਬǧǴƳ 2 ࢢյ಑ƟƧºȂǲǬÓǺÓఝƶǧǴǚࢪൟƳ‫ט‬СƟƧ‫ۼ‬¹഍෭ǚฤ঎ǣǺȉÓȦƳ 10 ൟյ।
Ɵ¹PBS Ư৽िƟƧºԇ໊ࣕ‫ۼ‬¹೪௾Ђஇ‫܎ڏ‬൝ࡐƶ๺ฤǚ Human Brain Mapping Analyzer Ư੯୩ƟƧº3.2܎ԙஇࡹ඲ƳǐǓऎ‫׫‬඲‫ۉ‬೰‫܎ڏ‬ൟ܎܎ஶ‫( ڐ‬SPM-RFL, Nikon) ƳǐƫƮ¹ॶ౗ƶ೪௾Ђஇ‫܎ڏ‬൝ࡐƶǴȕǪȃȦǚൟ৖Ɵ¹༐֬ౡସ‫ח‬ƻՒएȐǞȦǺÓǚଳ২ơǓƛưƳǐǓऎ‫׫‬඲ǚ‫ڸ‬௟ƟƧº3.3మࢲ੯܎඲ƳǐǓऎ‫׫‬඲ณ৾ॐƶ౗഍෭ǚ PBS Ư 2 ࠯յ৽िƟ¹঎ຜ঴ƶ‫܎ڏ‬൝ࡐǚࡷǒࣩƕ¹Human Brain Mapping AnalyzerƯԇ౗໹ЏƳƒƙǓ೪௾ЂஇƲ‫܎ڏ‬؊ஶǚ੯୩ƟƧº੶ƌƮ¹ৡඅǚ‫ื܎ڏ‬ћਭॎҠԙஇƳ৾ॐƟ¹ਗ਼૷ƶ௬Г‫ޕ‬഍ƳǶǾȃƟ¹௬Гऽ‫ڨ‬ҟƯ‫ޙ‬ƻ੯୩ƟƧºืћಶ҃‫ۼ‬ƶ‫܎ڏ‬؊ஶ૮ǐǒಶ҃ਕƶ૮ǚ‫ތ‬ƟСƕ¹࣐঑ƲืћਭॎҠԙஇ‫܎ڏ‬؊ஶ૮ǚ‫מ‬ǉƧáইŌâº௬Г‫ޕ‬഍Ƴ഍෭ǚǶǾȃơǓް¹ǴȤǟȄǧȤǴƶߡ૝ƳǧȤǴȕȮƯ௾୩ƶŎ஝ǚșÓǪƟƮ¹మஶ๋ƶ੯୩ƶ๋МưƟƧºŏ¼‫ื܎ڏ‬ћਭॎҠԙ৾ॐ๋இƶ‫ܨ‬ઑǚພƌƮǟǸǼǥǲǝȮ߱ȐȦǥȧǶǟȮ(FITC)ȤȔȦƶō‫ܨ‬ઑ඲ƳǐƫƮ¹‫ื܎ڏ‬ћਭॎҠԙஇƳ৾ॐƟƧºԇ‫ܨ‬ઑƶ౔ஶƷ¹ຊ೮ࡑ‫ۊ‬ƓǑ‫ڡ‬୩ƟƧºืћਭॎҠԙ৾ॐƶ੅߁ࡹࣔǚϱҟƳ࠷ơº(1) ഍෭ǚ PBS Ư ' ࠯յ৽िƟƧº(2) ഍࣍‫ڥ‬ঽǚৡඅƳƶƣ¹4ąƯ 2 ࠯յಶ҃ƝƣƧ‫ۼ‬¹PBSƯ 1 ࠯յ৽िƟƧº(3) ԇ൝ࡐƳઓơǓ‫ܨ‬ઑáફŌ‫ܨ‬ઑâǚ 1.2 ml ৡඅƳƶƣ¹4ąƯ 12 ࠯յಶ҃Ɲƣ
Ƨº(4) PBS Ư 30 ൟƢƬ 3 Ӗ৽िƟƧº(5) FITC Ư഍ࡀҠƝǔƧફō‫ܨ‬ઑǚৡඅƳƶƣ¹ǝȦȚȖǟȦইŌ.¸ॶ౗ৡඅƳ‫ۂ‬ǑǔǓ೪௾Ђஇ‫܎ڏ‬ƶࣩ‫׫‬඲ॶ౗ƳƷȥȘȐǴǼȮƲƱƳ֬ПơǓ೪௾ЂஇƲ‫܎ڏ‬൝ࡐƔൟരƟƮƒǒ¹‫ื܎ڏ‬ћਭॎҠԙஇƳ৾ॐƟƧ഍෭ƓǑ๋இươǓ൝ࡐƶൟരǚଽৣ௻ǓƛưƷƯƕƲƌºǮȥȮǝǶǼȦ֏஛Ѓ‫ݏ‬ਬáChATâƶൟ৖ǚ༎Ƴ¹ƥƶࣩ‫׫‬ධ඲ǚ࠷ơº৾ॐਕáAâư৾ॐ‫ۼ‬áBâƳ௬Гऽ‫ڨ‬ҟƯԇ໹Џƶ‫܎ڏ‬؊ஶǚ੯୩Ɵ¹ƥƶ‫ތ‬ǐǒ࣐঑Ʋ ChAT ƶ‫܎ڏ‬؊ஶ૮áCâǚ‫מ‬ǉƧºA/ॶ౗ৡඅƳƒƙǓ೪௾Ђஇ࠺ҫ‫܎ڏ‬ƶ؊ஶൟരই
B/ChAT ‫ܨ‬ઑƳǐǓืћਭॎҠԙ৾ॐ‫ۼ‬ƶ‫܎ڏ‬؊ஶൟരইºืћਭॎҠԙ৾ॐư࠺ҫ‫܎ڏ‬ƶൟരƔ‫ݜށ‬ƟƮƌǓºC/࣐঑Ʋ ChAT ƶ‫܎ڏ‬؊ஶƶൟരইºB Ƴ࠷ƟƧȂÓǺƓǑ A Ƴ࠷ƟƧȂÓǺǚ‫ތ‬СƗƛưƳǐƫƮ௻ǑǔǓº(J. Neurosci. Methods 85: 161-173, 1998 ǐǒСພ)-6-ইō#়ऺƲॶ౗ƳƒƙǓບÐƲ३‫ډ‬Ҡԙ൝ࡐƶൟരইƶГ༎৾ॐƟƧ౗഍෭ǚ Human Brain Mapping Analyzer ǚພƌƮ¹50 µmյԗƯ๢ 300 ญ໹ЏƳൟԥƟ¹ԇ໹ЏƳƒƙǓ‫ڏ‬܎؊ஶ૮ǚ੯୩ƟƧº௻ǑǔƧȂÓǺǚ 8 ૫ӭƶ؊ஶƳൟ་Ɵ¹ǦȤÓഒ࠷ƟƧºA/ൟ৖ƟƧ౗ৡඅƶ൉ϳ0Ыरƶ੃హইƳࠕ‫؁‬Ɣൟ৖ƟƧ໹Џǚ஝ਅƯ࠷ƟƧºƛƛƳ࠷ƟƧȂÓǺƶৡ૨฻ƷࡑਅƯ࠷ƟƧºAm!ኲொ0CC!౗໭0Cd!೯ीԌ0Cl!ਕम0CR!ද࡜Ն0DG!ࠦीӖ0FGi!ਕ௧ຢҟਕ௧Ӗ0FGm!ਕ
௧ຢଇਕ௧Ӗ0FGs!ਕ௧ຢरਕ௧Ӗ0GPe!૙੘‫ע‬ӱ২0GPi!૙੘‫ע‬ఝ২0Hp!ӥ౨0Hy!ࠜࣽҟ൉0IC!ఝඤਕ‫א‬0Pt!
೦ԍ0TGi!੩௧ຢҟ੩௧Ӗ0TGm!੩௧ຢଇ੩௧Ӗ0 TGs!੩௧ຢर੩௧ӖB/ǺǟȨǲȮ঎߱Ҡ‫ݏ‬ਬ (TH) ƶืћਭॎҠԙஇൟരইC/ǫȦǺȚȮ߱ે૛߱‫ݏ‬ਬ (GAD) ƶืћਭॎҠԙஇൟരইD/ǫȦǺȚȮ߱ે঎ਬ‫ݏ‬ਬ(GDH) ƶืћਭॎҠԙஇൟരই
E/ǰȑǴǺȮǴ P (SP) ƶืћਭॎҠԙஇൟരই
F/ǦȦȝǳȠȥȮ (CaM) ƶืћਭॎҠԙஇൟരই

(B&F: J. Neurosci. Res. 63: 369-376, 2001; C&D: Human Brain Mapping 11: 93-103, 2000; E: Neurosci. Res. 35: 339-346, 1999 ǐǒСພ)-7-Ưࡦ܎Ɵ¹ࡋҜƯ 3 ࠯յಶ҃ƝƣƧºϱ‫ۼ‬ƶ੅߁ƷਙƮࡦ܎ҟƯ݉ƲƫƧº(6) PBS Ư 3 ࠯յƢƬ 6 Ӗ৽िƟƧº(7) 10%ǫȥǶȥȮ PBS ຜњƯൊహƟƧº഍෭ǚ 4ąƶϬࣗƳඐըƟ¹ଽƩƳൟ৖ƟƧº‫܎ڏ‬ืћਭॎҠԙ৾ॐƶްƳ¹ȤǾȃƶ৛চৡඅǚॶ౗഍෭ư௬࠯Ƴ৾ॐƟ¹৾ॐऽ‫ڨ‬ƶࠉ഍ưƟƧº‫܎ڏ‬ืћਭॎҠԙ৾ॐƝǔƧ഍෭Ʒ¹ϬࣗୢҜҟƯГࢢյƷϩ୩ƯƊǓºGDH[14]¹ǰȑǴǺȮǴ P SP[13] ‫ח‬ƻǦȦȝǳȠȥȮCaM[16] ƶൟ৖ƶ‫ڤ‬ү¹ƥǔƦǔƶ൝ࡐƷ౗໹ЏƳϴ੻ƟƮൟരƟƮƒǒ¹ԇ൝ࡐƶൟരƳƷ‫ތ‬ЂƔǇǑǔƧº௾Ƴ¹೯ीԌ¹೦ԍ‫ח‬ƻ૙੘‫ע‬ƲƱƶ໹ЏƯ൝ࡐյƳ௾ଦஇƲൟരƔհߒƝǔƧºChAT Ʒ¹೦ԍ¹೯ीԌ¹૙੘‫ע‬¹ኲொ¹ਕमƲƱƶ໹ЏƳ‫ݗ‬ȧȔȦƯൟരƟƮƌƧºTH Ʒ೯ीԌư೦ԍƯ௾Ƴ‫ݗ‬ƌൟരƔ‫ۂ‬ǑǔƧºGAD ƶȧȔȦƷ૙੘‫ע‬ưࠜࣽҟ൉Ư‫ޚ‬Ǌ‫ݗ‬Ɨ¹૙੘‫ע‬ƶӱ২ǐǒǊఝ২Ƴ‫ݗ‬ƌൟരƔ‫ۂ‬ǑǔƧºGDH ƷӦಋࡐਙઑƳೝԔஇ‫ج‬Г
Ő¼୩໷ൟ৖ƳൟരƟƮƌƧƔ¹೦ԍ¹೯ीԌ¹૙੘‫ע‬¹ࠜࣽҟൟ৖ƳƷ Human Brain Mapping Analyzer [9]ǚພƌƧº ൉ƲƱƳ‫ݗ‬ȧȔȦƯ੻޲ƟƧºSP Ʒ૙੘‫ע‬ƶఝ২ƛƶਗ਼૷Ʒົ࡜‫ۉ܎ڏ‬೰‫؛‬ƶǦȜȤșǡȮȃƳ‫ݗ‬ՕƳƶǇ‫ݗ‬ȧȔȦƯ‫޲ؤ‬ƟƮƌƧºCaM ƷӦಋࡐƶਙஶƶ܎஢ࠃ੢౵ըǚǶǾȃƟƮƊǒ¹ȏȮȖÓȦǚƮƶ໹ЏƯೝԔஇ‫ݗ‬ƌൟരǚ࠷ƟƧƔ¹೯ीԌư೦ୋƟƮ¹഍෭ƶ೰ެƲ໹ЏƓǑƶ௾୩Ʋ‫܎ڏ‬؊ஶǚԍƶȧȔȦƔ‫ޚ‬Ǌ‫ݗ‬ƓƫƧº෭඲ƳǐƫƮ௻ǑǔƧ‫ڐ‬੯ơǓਗ਼૷ƯƊǓº‫ۉ‬೰‫؛‬ƶǴȁÓǳƷǮȮȏȠ‫ڤ‬үƷϱਕƶিҠԙஇƲ‫ڤ‬ү‫ח‬ƻਭॎҠԙஇհߒ‫ڤ‬ÓǺÓƯর‫۽‬ƝǔƮƒǒ¹഍෭ਙ฻ƶ‫܎ڏ‬؊ஶƶൟүưઓ҃ƟƮƒǒ¹়ԏƲൟരư୩໷૮ǚެභȧȔരǚެභȧȔȦƯГஶƳൟ৖ƯƕǓºǴȁÓǳƶ‫ޚ‬ȦƯ௻ǓƛưƔƯƕƧºપҧ௫ೆ϶Ʒ 150 x 150 mm¹‫ࣹޚ‬੯܎յԗƷ 0.25 µm¹‫ݶ‬Ӗԏ໔ƟƧൟ৖ධ඲Ʒ¹ࢫີƶධ඲ưೝԔƟƮ੯܎ǴȏÓȄƷ 100 ໹ЏÖഘƯƊǓºଙƟƗϩ୩ƟƧ়ԏƲ‫ڤ‬үƔ௻ǑǔǓºƛƎƟƮ௻੯୩ऽ‫¸¾ڨ‬༐֬ౡସ 420-490 nm, ՒएȐǞȦǺǑǔǓൟ৖‫ڤ‬үƷ¹ଇঞౖ֞ƶӕฯǌࡾÐƶ३‫ࡏډ‬
Ó 530 nm, ܎஢ࠃ੢౵ըƶМҦ஢Ϝ 850 V, ઓ൝ȧՔƶದऔ֞ࣥӕฯƶƧǉƶ֏ਨஇȂÓǺǚ୲‫׼‬ơǓ
Ȯǵ 20 x / 0.5 (౵໓/Ӭ‫ܔ‬ঝ), ੯܎յԗ 50 µm, ࠜ๝ǊƶƯƊǓºܿǒ 40 µm, ੯܎ܿǒ 100 µmº෭‫ףڽ‬ƯƷ¹1 ෸ƶ౗഍෭ƓǑ๢ 300 ญ໹ЏƶȂÓǺǚ௻Ƨº௻ǑǔƧ‫܎ڏ‬؊ஶ૮ǚ 8 ૫ӭƳǦȤÓഒ࠷Ɵ¹ԇ൝ࡐƶൟരǚӕ৖ƟƧºǡȤȮǧȤǴǚ‫܎ڏ‬؊ஶƶǴǺȮǻÓ
ߣ݂൫‫ڼ‬
ȄưƟƮ߹ພƟƧº[1] অ௞ஞў, ࢖ߤҥન, ਕ஡Гགྷ. ౗ఝ३‫ډ‬ஞૄ൝ࡐൟരൟ৖ƶƧǉƶ‫ۉ‬೰੯܎ਗ਼૷ǲǴȁțƶ
ő¼‫ڤ‬үư݂ߒӬದ, ష෭๣໊ԙߙࠠ 91 (1988) 173-180.
‫ݗ‬༛ƶॶ౗ƳƷȥȘȐǴǼȮ௖Ƴ຀ີơǓ࠺ҫ‫ڏ‬[2] D. Sutoo, K. Yabe, K. Akiyama, Atlas of the rat
brain: quantitative distribution of the choline܎Ɣ੻޲ơǓƧǉ¹ࣹ௫൝ƶ౗഍෭ƶǐƎƳଽৣ¹acetyltransferase, Kitasato Arch. Exp. Med. 64ൟ৖ơǓƛưƔࢽີƲƌº೪௾Ђஇ࠺ҫ‫܎ڏ‬Ʒ¹๺
(1991) 221-262.
֞ຜ౷¹߱½ǝȦǦȥ¹ӄ߱Ҡ঎ਬ঎ƳǐǓ߱Ҡࣕ
[3] D. Sutoo, K. Akiyama, K. Yabe, K. Kohno, Multiple
໊¹঎ਬҠǝȦȚȆǡțȥǼǡțǌ঎ਬҠȖǡਬȅ
analysis of tyrosine hydroxylase and calmodulin
distributions in the forebrain of the rat using aȃȥǡțƳǐǓճ‫໊ࣕی‬ƯƷ¹ǃưǛƱऎ‫׫‬ƯƕƲ
microphotometry system, Brain Res. Bull. 26 (1991)
ƓƫƧºӄșȮǧȮ߱ǦȥǡțƳǐǓ߱Ҡ໊ࣕƷ¹973-982.
೪௾Ђஇ࠺ҫ‫܎ڏ‬ƶ؊ஶǚ๢ 50 % ‫ࣺۑ‬ƝƣƧƔ¹[4] D. Sutoo, K. Akiyama, K. Yabe, N. Nishida,ਭॎ഍෭Ɣ୊Ǉ¹ืћਭॎҠԙ৾ॐƳ߹ພơǓࠧƔ
Quantitative immunohistochemical distribution ofcholine acetyltransferase in the rostral forebrain ofƯƕƲƓƫƧºѻਬǧǴƳǐǓ߱Ҡഏಋಶ҃Ʒ࠺ҫ
the rat, Neurosci. Res. 11 (1991) 63-70.
‫܎ڏ‬ƶ؊ஶǚ๢ 90 % ‫ࣺۑ‬ƝƣƧƔ¹‫۔ޙ‬঴Ƴ๓ભ
[5] K. Akiyama, K. Yabe, D. Sutoo, QuantitativeƔƊƫƧºƝǑƳ¹࠺ҫ‫܎ڏ‬Ʒ‫ܢ‬ƌౡସ໹Џƶ༐֬
immunohistochemical distributions of tyrosine܎Ư༐֬Ɲǔ¹ದƣǑǔǓ‫܎ڏ‬Ǌ‫ܢ‬ƌౡସ໹ЏƳ஬hydroxylase and calmodulin in the brains of
spontaneously hypertensive rats, Kitasato Arch. Exp.
ǓƧǉ¹༐֬ౡସƶਏવǌՒएȐǞȦǺÓƳǐƫƮ¹Med. 65 (1992) 199-208.
ืћਭॎҠԙஇ‫܎ڏ‬ư࠺ҫ‫܎ڏ‬ǚൟ໐ơǓƛưƷƯ
[6] M. Hanawa, T. Asano, K. Akiyama, K. Yabe, K.
ƕƲƓƫƧºTsunoda, T. Tadano, D. Sutoo, Effect of Zena F-III, a
ƥƛƯ¹Human Brain Mapping Analyzer ǚພƌƮliquid nutritive and tonic drug, on the neurochemicalchanges elicited by physical fatigue in mice,ืћਭॎҠԙ৾ॐਕ‫ۼ‬Ƴ௬Гऽ‫ڨ‬ҟƯ௬Ơ൉ϳƶ‫ڏ‬
Pharmacol. Biochem. Behav. 66 (2000) 771-778.
܎؊ஶǚ 2 Ӗ੯܎Ɵ¹ಶ҃‫ۼ‬ƶ૮ƓǑಶ҃ਕƶ૮ǚ[7] D. Sutoo, K. Akiyama, K. Yabe, A novel technique‫ތ‬ƟСƗమࢲ੯܎඲ǚພƌƮ࣐঑Ʋ‫܎ڏ‬؊ஶ૮ǚ௻
for quantitative immunohistochemical imaging ofƧºƛƶධ඲Ʒ¹Ҡԙஇಶ҃ǚພƌƧऎ‫׫‬඲ưЂƲvarious neurochemicals in a multiple-stained brainslice, J. Neurosci. Methods 118 (2002) 41-50.
ǒ¹‫܎ڏ‬ƶણॐǌਭॎ഍෭ƶ୊ǇǊǃưǛƱฤƗ¹[8] D. Sutoo, K. Akiyama, K. Yabe, Comparison‫ޚ‬Ǌ়ԏƯ‫۔ޙ‬঴ƔƊƫƧáইŌâ [9]ºƛƶධ඲Ƴ
analysis of distributions of tyrosine hydroxylase,ǐƫƮ௻ǑǔƧԇ൝ࡐƶ࣐঑Ʋ‫܎ڏ‬؊ஶƶൟരǚই
calmodulin
andcalcium/calmodulin-dependentŌ‫ח‬ƻই 2 Ƴ࠷ƟƮƌǓºprotein kinase II in a triple stained slice of rat brain,Brain Res. 933 (2002) 1-11.
ǮȥȮǝǶǼȦ֏஛Ѓ‫ݏ‬ਬǝǶǼȦǮȥȮƶ‫ݜ‬[9] D. Sutoo, K. Akiyama, K. Yabe, Quantitative঵‫ݏ‬ਬ! ChAT[9,10] ¹ǺǟȨǲȮ঎߱Ҡ‫ݏ‬ਬǦȁǮ
mapping analyzer for determining the distribution ofȦǝȚȮƶ‫ݜ‬঵‫ݏ‬ਬ! TH[10,16]¹ǫȦǺȚȮ߱ે૛߱
neurochemicals in the human brain, J. Neurosci.
[14]‫ݏ‬ਬGABA ƶ‫ݜ‬঵‫ݏ‬ਬ! GAD ¹ǫȦǺȚȮ߱ે
Methods 85 (1998) 161-173.
঎ਬ‫ݏ‬ਬǫȦǺȚȮ߱ƶȧȔȦǚଳ২ơǓ‫ݏ‬ਬ!

-8-

[10] D. Sutoo, K. Akiyama, K. Yabe, K. Kohno,
Quantitative analysis of immunohistochemical
distributions of cholinergic and catecholaminergic
systems in the human brain, Neuroscience 58 (1994)
227-234.
[11] K. Yabe, D. Sutoo, K. Akiyama, K. Kohno,
Quantitative immunohistochemical distributions ofsome neurotransmitters and related enzymes in thehuman brain, XIIth International Congress ofPharmacology (Montréal, Canada), 1994.
[12] K. Yabe, K. Akiyama, D. Sutoo, Quantitative
immunohistochemical distributions of various
neurochemicals in the human brains, XIIIthInternational Congress of Pharmacology (München,Germany), 1998.
[13] D. Sutoo, K. Yabe, K. Akiyama, Quantitative
imaging of substance P in the human brain using abrain mapping analyzer, Neurosci. Res. 35 (1999)
339-346.

-9-[14] D. Sutoo, K. Akiyama, K. Yabe, Quantitative mapsof GABAergic and glutamatergic neuronal systems
in the human brain, Human Brain Mapping 11 (2000)
93-103.
[15] D. Sutoo, K. Yabe, K. Akiyama, Quantitative
distribution of various neurochemicals in the humanbrain using a human brain mapping analyzer, In: T.
Kato (Ed.), Frontiers of the Mechanisms of Memory
and Dementia, Elsevier, Amsterdam, 2000, pp. 4546.
[16] D. Sutoo, K. Akiyama, K. Yabe, Quantitative
imaging of tyrosine hydroxylase and calmodulin inthe human brain, J. Neurosci. Res. 63 (2001) 369376.