ᑐࡍࡿ⚄⤒㞀ᐖᛶࡀ♧၀ࡉࢀࡓࠋ

࢔ࢡࣜࣝ࢔࣑ࢻࡢࣛࢵࢺ࡬ࡢⓎ㐩ᮇᭀ㟢ᐇ㦂࡟࠾࠸࡚ , ẕື≀࡟⚄⤒ẘᛶࢆ♧ࡍ⏝㔞ࡢ

36

ᭀ㟢࡟ࡼࡗ࡚ࡶᏊື≀࡟ᑐࡋ࡚᫂ࡽ࠿࡞㍈⣴㞀ᐖࡣࡳࡽࢀ࡚࠸࡞࠸ࡀ㸦 87 㸧 , Ꮚື≀ࡢ SGZ

࡛ᮍ⇍㢛⢏⣽⬊ࢆᶆⓗ࡜ࡋࡓࢽ࣮ࣗࣟࣥ᪂⏕ࡢᚋᮇ㐣⛬ࡢ㞀ᐖࡀ᳨ฟࡉࢀ࡚࠸ࡿ㸦 61, 62 㸧 ࠋ ᮏ❶࡟࠾ࡅࡿࢢࣜࢩࢻ࣮ࣝࡢⓎ㐩ᮇᭀ㟢ࡣ, ⏕ᚋ 21 ᪥ࡢ 1,000 ppm ⩌࡛ᾏ㤿ṑ≧ᅇ SGZ ࡢ

TUC-4 ࡢⓎ⌧ศᕸ࡟ᙳ㡪ࢆཬࡰࡍࡇ࡜ࡀ☜ㄆࡉࢀࡓࠋTUC-4 ࡣᮍ⇍㢛⢏⣽⬊࡛Ⓨ⌧ࡍࡿࡇ

࡜ࡀ▱ࡽࢀ࡚࠸ࡿࡇ࡜࠿ࡽ㸦35㸧, ᮍ⇍㢛⢏⣽⬊ࡢῶᑡࡀ♧၀ࡉࢀࡓࠋGFAP ࡣ type-1 ⚄⤒

ᖿ⣽⬊࡛ , Pax6 ࡣ type-1 ⚄⤒ᖿ⣽⬊࠿ࡽ type-2a ๓㥑⣽⬊ࡲ࡛࡛ , Tbr2 ࡣ type-2b ๓㥑⣽⬊࡛ ,

ࡲࡓ , DCX ࡣ type-2b ๓㥑⣽⬊࠿ࡽᮍ⇍㢛⢏⣽⬊ࡲ࡛࡛Ⓨ⌧ࡍࡿศᏊ࡛࠶ࡾ㸦 26, 34 㸧 , ࡇࢀ

ࡽ๓ᮇࢽ࣮ࣗࣟࣥ᪂⏕ࡢᣦᶆࡢኚືࡣㄆࡵࡽࢀ࡞࠿ࡗࡓࠋ ᮍ⇍㢛⢏⣽⬊ࡣ᪤࡟⚄⤒✺㉳ᡂ㛗

෇㗹ཬࡧᇶᗏᶞ≧✺㉳ࢆ᭷ࡋ࡚࠸ࡿ㸦73㸧 ࠋࡲࡓ, ⚄⤒ࡢⓎ㐩㐣⛬࡟࠾ࡅࡿࢽ࣮ࣗࣟࣥ⛣ື

ࢆไᚚࡍࡿศᏊᶵᗎࡣ, ㍈⣴࢞࢖ࢲࣥࢫࡸ⚄⤒✺㉳ᙧᡂ࡜㢮ఝᛶࡀㄆࡵࡽࢀ࡚࠸ࡿ㸦58㸧 ࠋ ௨ୖࡢࡇ࡜࠿ࡽ , ᡂ⇍ື≀࡛ࡢ㍈⣴㞀ᐖᛶࢆ⪃៖ࡍࡿ࡜ , ࢢࣜࢩࢻ࣮ࣝࡀⓎ㐩⚄⤒ẘ࡜ࡋ࡚

᪂⏕⚄⤒ᮎ➃࡟┤᥋ᙳ㡪ࡍࡿࡇ࡜࡛ᮍ⇍㢛⢏⣽⬊ࡢศ໬ࢆ㞀ᐖࡍࡿࡇ࡜ࡀ᥎ ࡉࢀࡓࠋ ࡋ࠿

ࡋ࡞ࡀࡽ , ᾏ㤿ṑ≧ᅇ࡟࠾ࡅࡿࡇࢀࡽࡢኚ໬ࡣ⏕ᚋ 77 ᪥࡛ࡣㄆࡵࡽࢀࡎ , ྍ㏫ᛶࡢኚ໬࡛

࠶ࡗࡓࠋ

୍᪉, ᾏ㤿ṑ≧ᅇ㛛࡛ࡣ࢔ࢡࣜࣝ࢔࣑ࢻࡸᢠ⏥≧⭢๣ࡢⓎ㐩ᮇᭀ㟢ᐇ㦂ࡢሗ࿌㸦62, 76,

81㸧࡜ྠᵝ, reelin 㝧ᛶ⣽⬊, Calb2 㝧ᛶ⣽⬊ཬࡧ NeuN 㝧ᛶ⣽⬊ࡢቑຍࡀ⏕ᚋ 21 ᪥ࡢ 1,000

ppm ⩌࡛☜ㄆࡉࢀࡓࠋṑ≧ᅇ㛛ࡢ Calb2 㝧ᛶ௓ᅾࢽ࣮ࣗࣟࣥࡣ , 㢛⢏⣽⬊࡟ᢞᑕࡍࡿ௚ࡢ GABA ᛶࢽ࣮ࣗࣟࣥ࡜┦஫࡟㐃⤡ࡋ࡚άᛶࢆไᚚࡍࡿࡇ࡜࡛ , ᾏ㤿඲యࡢάᛶࢆㄪᩚࡍࡿ

㸦23㸧 ࠋࡲࡓ, reelin ࡣ GABA ᛶ௓ᅾࢽ࣮ࣗࣟࣥ࠿ࡽศἪࡉࢀ, ᾏ㤿ṑ≧ᅇ࡟࠾࠸࡚⏕ᚋࡢࢽ

࣮ࣗࣟࣥ᪂⏕ࢆㄪ⠇ࡍࡿᙺ๭ࢆᢸࡗ࡚࠸ࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ㸦10, 21, 47, 72㸧 ࠋࡋࡓࡀࡗ

࡚, GABA ᛶ௓ᅾࢽ࣮ࣗࣟࣥ࡟࠾ࡅࡿ, 㢛⢏⣽⬊ࡢ᪂⏕ࡢ㞀ᐖ࡜ࡑࢀ࡟ᘬࡁ⥆ࡃ⚄⤒⛣ື㞀 ᐖࢆ཯ᫎࡍࡿኚືࢆ᳨ฟࡋࡓࡶࡢ࡜⪃࠼ࡽࢀࡿࠋࡉࡽ࡟, reelin 㝧ᛶ௓ᅾࢽ࣮ࣗࣟࣥࡣ NeuN 㝜ᛶࡲࡓࡣᙅ㝧ᛶ࡛࠶ࡗࡓࠋ NeuN ࡢపⓎ⌧ࡣᮍ⇍࡞ࢽ࣮࡛ࣗࣟࣥ࠶ࡿࡇ࡜ࢆ♧ࡋ࡚࠾ࡾ

㸦 78 㸧 , TUC-4 㝧ᛶᮍ⇍㢛⢏⣽⬊ࡢῶᑡࢆ཯ᫎࡋ࡚ reelin ࢆⓎ⌧ࡍࡿᮍ⇍ࢽ࣮ࣗࣟࣥࡢቑຍ

ࢆᘬࡁ㉳ࡇࡍࡶࡢ࡜᥎ ࡉࢀࡓࠋᮍ⇍㢛⢏⣽⬊ᩘࡢኚືࡣ⏕ᚋ 21 ᪥ࡢࡳ࡛࠶ࡗࡓࡀ , ௓ᅾ

37

ࢽ࣮ࣗࣟࣥࡢኚືࡣᡂ⇍ࡲ࡛⥆ࡃࡶࡢ࡛࠶ࡿࡇ࡜ࡀ☜ㄆࡉࢀ , ࢽ࣮ࣗࣟࣥ᪂⏕ࡸ⛣ື㞀ᐖ ࡢ⥅⥆ᛶࢆ♧ࡍࡶࡢ࡜᥎ ࡉࢀࡓࠋ

⏥≧⭢㛵㐃࣍ࣝࣔࣥࡣ⬻ࡢⓎ㐩࡟㔜せ࡛࠶ࡿ 㸦3㸧 ࠋ ᮏ❶࡛ࡣ, T

3

ࡢቑຍࡀ⏕ᚋ 21 ᪥ࡢ 1,000 ppm ࡢᏊື≀࡛ࡳࡽࢀࡓࠋᡂ⇍ࣛࢵࢺ࡛ࡢ⏥≧⭢ᶵ⬟ஹ㐍ࡣࢩࢼࣉࢫྍረᛶࡸグ᠈㞀ᐖࢆ

ᘬࡁ㉳ࡇࡍࡇ࡜࠿ࡽ㸦89㸧, ࢽ࣮ࣗࣟࣥ᪂⏕࡟ࡶᙳ㡪ࢆཬࡰࡍྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋ1,000 ppm ⩌ࡢᏊື≀࡟࠾࠸࡚ࡶ , ࢽ࣮ࣗࣟࣥ᪂⏕࡟ᙳ㡪ࢆ୚࠼ࡿ⏥≧⭢࣍ࣝࣔࣥࡢ␗ᖖࡀㄏⓎ

ࡉࢀࡓྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࡀ , ẕື≀࡛ࡣ T

3

ཬࡧ T

4

⃰ᗘ࡬ࡢᙳ㡪࡞ࡋ࡟㍈⣴㞀ᐖࡀㄏⓎࡉ

ࢀࡓࡇ࡜࠿ࡽࡶ , ࢢࣜࢩࢻ࣮ࣝ⮬యࡢ㍈⣴㞀ᐖᛶࡀᏊື≀࡟࠾ࡅࡿ᪂⏕⚄⤒ᮎ➃ࡢ㞀ᐖᛶ

ࢆ௓ࡋࡓࢽ࣮ࣗࣟࣥ᪂⏕㞀ᐖࢆㄏⓎࡋࡓ࡜᥎ ࡉࢀࡿࠋ

୍᪉, ⏕ᚋ 21 ᪥ࡢᏊື≀ࡢᾏ㤿ṑ≧ᅇ࡟࠾ࡅࡿ㑇ఏᏊⓎ⌧ゎᯒ࡟࠾࠸࡚, 1,000 ppm ⩌࡛

୺࡟

Ncam2, Nfasc, Mrf, Mal

ཬࡧ

Ugt8

࡞࡝⚄⤒ྍረᛶ , 㧊㠧ᙧᡂ࡟㛵୚ࡍࡿ㑇ఏᏊ⩌ 㸦 14, 36,

37, 77, 94 㸧ࡢⓎ⌧㔞ῶᑡࡀ☜ㄆࡉࢀࡓࠋࡇࢀࡽࡣ , ᾏ㤿ṑ≧ᅇ SGZ ࡛ࡢࢽ࣮ࣗࣟࣥ᪂⏕ࡢ

ᚋᮇ࡟࠾ࡅࡿ㍈⣴ࡸ⚄⤒✺㉳ࡢఙᙇάᛶࢆ᭷ࡍࡿᮍ⇍㢛⢏⣽⬊ࡢῶᑡ࡜㛵㐃ࡋࡓ , ⚄⤒✺

㉳ࡢᡂ㛗ࡸࡑࢀ࡟⥆ࡃ⚄⤒ᅇ㊰ᙧᡂࡢᢚไࢆ♧၀ࡍࡿኚື࡜⪃࠼ࡽࢀࡿࠋࡲࡓ, ᾏ㤿ṑ≧ᅇ

࡟࠾ࡅࡿච␿⤌⧊໬Ꮫⓗゎᯒ࡟ࡼࡗ࡚ PCNA 㝧ᛶቑṪ⣽⬊࡟ኚືࡣࡳࡽࢀ࡞࠿ࡗࡓࡀ, 㑇 ఏᏊⓎ⌧ゎᯒ࡟ࡼࡗ࡚ 1,000 ppm ⩌࡛

Aurkb, Ndc80

ཬࡧ

Cdk1

࡞࡝ࡢ⣽⬊ቑṪ㛵㐃㑇ఏᏊ 㸦9, 75 㸧ࡢⓎ⌧㔞ῶᑡࡀ☜ㄆࡉࢀ , ⣽⬊࿘ᮇࡢᢚไࡀ♧၀ࡉࢀࡓࠋ⣽⬊ቑṪάᛶ㜼ᐖࡢᶵᗎࡣ᫂

ࡽ࠿࡛࡞࠸ࡀ , ࢽ࣮ࣗࣟࣥ᪂⏕㞀ᐖࢆ཯ᫎࡋࡓኚື࡛࠶ࡿ࡜᥎ ࡉࢀࡿࠋᖏ≧ᅇ࡛ࡣ

Tnc, Cd44

ཬࡧ

Efnb3

࡞࡝ࡢ㍈⣴ᙧᡂ㛵㐃㑇ఏᏊ㸦31, 42, 53㸧ࡢⓎ⌧㔞ቑຍࡀ, ᑠ⬻⓶㉁࡛ࡣ

Ccl2

ཬࡧ

Thbs2

࡞࡝ࡢࢩࢼࣉࢫ㛵㐃㑇ఏᏊ㸦7, 100㸧ࡢⓎ⌧㔞ῶᑡࡀࡳࡽࢀࡓࠋࡇࢀࡽࡶࢢࣜࢩ

ࢻ࣮ࣝᭀ㟢࡟ࡼࡿ⚄⤒ྍረᛶ࡬ࡢᙳ㡪ࢆ཯ᫎࡍࡿࡶࡢ࡜♧၀ࡉࢀࡓࠋࡲࡓ, ⬻ᱱ࡛ࡣ, Kl,

Igf2

ཬࡧ

Igfbp2

࡞࡝ࡢ㧊㠧ᙧᡂ࡟㛵ࢃࡿ㑇ఏᏊ㸦8, 38㸧ࡢⓎ⌧㔞ῶᑡࡀࡳࡽࢀ, ㍈⣴ᙧᡂ, ࢩ ࢼࣉࢫᙧᡂ࡬ࡢᙳ㡪ࢆ཯ᫎࡋࡓ㧊㠧㛵㐃㑇ఏᏊࡢኚື࡜ࡋ᳨࡚ฟࡉࢀࡓࡶࡢ࡜♧၀ࡉࢀࡓࠋ

ྛ⬻㒊఩࡟࠾ࡅࡿⓎ⌧ኚື㑇ఏᏊࡢ࠺ࡕ , Cntn3 ࡣ 1,000 ppm ⩌࡛ 4 ࡘࡢ⬻㒊఩࡟ඹ㏻ࡋ࡚ୖ

᪉ไᚚࡉࢀࡓࠋࡇࢀࡣ⚄⤒᥋╔ศᏊ࡜ࡋ࡚▱ࡽࢀ , ⚄⤒✺㉳ᡂ㛗άᛶࢆ᭷ࡋ , ṑ≧ᅇ㢛⢏⣽

38

⬊ , ᑠ⬻ࣉ࢚ࣝ࢟ࣥ⣽⬊ཬࡧ኱⬻⓶㉁ࡢ⾲ᒙ࡟ศᕸࡍࡿࢽ࣮ࣗࣟࣥ࡞࡝࡛Ⓨ⌧ࡋ࡚࠾ࡾ

㸦 97 㸧 , ࢢࣜࢩࢻ࣮ࣝⓎ㐩ᮇᭀ㟢࡟ࡼࡿẘᛶࢆ⌧ࡍᶆⓗ㑇ఏᏊ࡜ࡋ࡚࠶ࡆࡽࢀࡿ࡜⪃࠼ࡽࢀ

ࡓࠋࡇࢀࡽࡢᾏ㤿ṑ≧ᅇ࡛ኚືࡀࡳࡽࢀࡓ㑇ఏᏊ⩌ࡢ࠺ࡕ, Nfasc, Mrf, Mal, Aukb ཬࡧ

Ndc80

࡞࡝ከࡃࡢ㑇ఏᏊࡣ 300 ppm ⩌࡛ࡶྠᵝࡢኚືࡀࡳࡽࢀࡓࡀ, ௚ࡢ 3 㒊఩࡛ࡣ࡯࡜ࢇ࡝ࡀ

1,000 ppm ⩌ࡢࡳࡢኚື࡛࠶ࡗࡓࠋᾏ㤿ṑ≧ᅇࡣᡂ⇍ᚋࡲ࡛ࢽ࣮ࣗࣟࣥ᪂⏕ࢆ⥆ࡅࡿࡇ࡜࠿

ࡽ , 4 㒊఩ࡢ୰࡛㑇ఏᏊⓎ⌧ゎᯒ࡟ࡼࡿⓎ㐩⚄⤒ẘᛶᙳ㡪ࡢ᳨ฟ࡟㛵ࡋ࡚᭱ࡶឤཷᛶࡢ㧗࠸

㒊఩࡛࠶ࡿࡇ࡜ࡀ♧၀ࡉࢀࡓࠋᮏᐇ㦂࡟࠾࠸࡚ , 1,000 ppm ⩌ࡢẕື≀ࡢ୰ᯡ⚄⤒࡛ࡣᑠ⬻

ཬࡧᘏ㧊⫼⣴࡛ࢫࣇ࢙ࣟ࢖ࢻᙧᡂࡀぢࡽࢀࡓࡀ , Ꮚື≀ࡢ⬻࡛ࡣ , NF-L ཬࡧ beta III tubulin

ࡢච␿ᰁⰍ࡟ࡼࡿᙧែⓗホ౯࡛ࡣ㍈⣴ཬࡧ⚄⤒✺㉳ࡢ㞀ᐖࡣ᫂ࡽ࠿࡛࡞࠿ࡗࡓࠋࡋ࠿ࡋ, ≉

ᐃ⬻㒊఩࡟࠾ࡅࡿ㑇ఏᏊⓎ⌧ゎᯒ࡟ࡼࡗ࡚⋓ᚓࡋࡓ㍈⣴ᙧᡂࡸࢩࢼࣉࢫᙧᡂ࡟㛵㐃ࡍࡿ㑇

ఏᏊࢡࣛࢫࢱ࣮ࡣ , Ⓨ㐩ᮇᭀ㟢࡟ࡼࡿ⚄⤒ྍረᛶ࡬ࡢᙳ㡪࡟㛵㐃ࡍࡿࡶࡢ࡜♧၀ࡉࢀࡓࠋ

39 ᑠ ᑠ ᣓ

➨ 2 ❶࡛ࡣ, ࢢࣜࢩࢻ࣮ࣝࢆࣔࢹࣝ≀㉁࡜ࡋ࡚⏝࠸࡚, ᡂ⇍ື≀ࡢ⚄⤒㞀ᐖཬࡧᏊື≀ࡢ ᾏ㤿ṑ≧ᅇ࡟࠾ࡅࡿࢽ࣮ࣗࣟࣥ᪂⏕࡬ࡢᙳ㡪࡟ࡘ࠸᳨࡚⣴ࡋ, ࡑࢀࡽࡢᶆⓗᛶ࡟ᑐࡍࡿẘ ᛶⓎ⌧ᶵᗎࡢඹ㏻ᛶࢆ᳨ウࡋࡓࠋࡲࡓ, Ꮚື≀ࡢྛ⬻㒊఩࡟࠾ࡅࡿ㑇ఏᏊⓎ⌧ࣉࣟࣇ࢓࢖ࣝ

ࢆ⋓ᚓࡋࡓࠋ

ࢢࣜࢩࢻ࣮ࣝࡢࣛࢵࢺࢆ⏝࠸ࡓⓎ㐩ᮇᭀ㟢࡛ , 108.8 mg/kg body weight/day ࡟┦ᙜࡍࡿ

1,000 ppm ࡢࣞ࣋ࣝ࡟࠾࠸࡚ , ẕື≀ࡢ୰ᯡཬࡧᮎᲈ⚄⤒⣔࡟㍈⣴㞀ᐖࢆㄏⓎࡍࡿࡇ࡜ࢆぢ

ฟࡋࡓࠋࡲࡓ, ྠ⏝㔞࡟࠾࠸࡚, ྍ㏫ⓗ࡞ࡀࡽᏊື≀ࡢᾏ㤿ṑ≧ᅇ SGZ ࡛ࡢศ໬ᚋᮇ࡟࠾ࡅ

ࡿ⚄⤒✺㉳ᙧᡂࢆᶆⓗ࡜ࡋࡓ, ࢽ࣮ࣗࣟࣥ᪂⏕ࡢᚋᮇ㐣⛬࡬ࡢᙳ㡪ࢆぢฟࡋࡓࠋ

Ꮚື≀ࡢ⬻㒊఩ูࡢ㑇ఏᏊⓎ⌧ゎᯒ࡛ࡣ , ୺࡟㍈⣴ᙧᡂ , ⚄⤒✺㉳ᙧᡂ , 㧊㠧ᙧᡂཬࡧࢩ ࢼࣉࢫ㛵㐃㑇ఏᏊࡢⓎ⌧㔞ࡢኚືࡀホ౯࡟⏝࠸ࡓ⬻ 4 㒊఩࡛☜ㄆࡉࢀ , ᾏ㤿ṑ≧ᅇ SGZ ࡛ ぢฟࡋࡓ⚄⤒✺㉳ᙧᡂ࡬ࡢᶆⓗᛶ࡜㛵㐃ࡋࡓ⚄⤒ᅇ㊰ࡢྍረᛶ࡬ࡢᙳ㡪ࢆ♧ࡍ㑇ఏᏊࢡࣛ

ࢫࢱ࣮࡜ࡋ᳨࡚ฟࡉࢀࡓࡶࡢ࡜⪃࠼ࡽࢀࡓࠋࡲࡓ, ᾏ㤿ṑ≧ᅇ࡛ࡣ 300 ppm ⩌࡟࠾࠸࡚ࡶ

1,000 ppm ⩌࡜ྠᵝࡢ㑇ఏᏊⓎ⌧ኚືࡀ☜ㄆࡉࢀࡓࡀ, ௚ࡢ 3 㒊఩࡛ࡣ࡯࡜ࢇ࡝ࡢ㑇ఏᏊࡀ

1,000 ppm ⩌ࡢࡳࡢኚື࡛࠶ࡗࡓࠋࡇࡢࡇ࡜ࡣ, ᡂ⇍ᚋࡶࢽ࣮ࣗࣟࣥ᪂⏕ࢆ⥆ࡅࡿᾏ㤿ṑ≧

ᅇࡀ , ᮏ≀㉁ࡢ⚄⤒ẘᛶࡢᶆⓗ࡜ࡋ࡚᭱ࡶឤཷᛶࡢ㧗࠸㒊఩࡛࠶ࡿࡇ࡜ࡀ᥎ ࡉࢀࡓࠋ

ୖ㏙ࡢࡼ࠺࡟ , ᮏ◊✲࡟ࡼࡾࢢࣜࢩࢻ࣮ࣝ࡟ࡼࡿᡂ⇍ື≀࡟࠾ࡅࡿ㍈⣴ኚᛶ࡜Ⓨ㐩୰ື

≀ࡢࢽ࣮ࣗࣟࣥ᪂⏕ࡢᚋᮇ㐣⛬ࢆᶆⓗ࡜ࡋࡓⓎ㐩⚄⤒ẘᛶࡀぢฟࡉࢀ, ྜࢃࡏ࡚Ꮚື≀ࡢ

⬻࡟࠾ࡅࡿ⚄⤒ᅇ㊰ࡢྍረᛶ࡬ࡢᙳ㡪ࢆ♧ࡍ㑇ఏᏊࢡࣛࢫࢱ࣮ࡢ⋓ᚓ࡟ࡼࡾ, ࢢࣜࢩࢻ࣮

ࣝࡢẘᛶᶵᗎ࡜ࡋ࡚, ᡂ⇍ࡋࡓ㍈⣴࡜Ⓨ㐩୰ࡢ⚄⤒✺㉳࡟ᑐࡋ࡚ඹ㏻ࡢᶆⓗᛶࢆᣢࡘࡇ࡜

ࡀ᥎ ࡉࢀࡓࠋ

40

➨

➨ 3 ❶

ࢢࣜࢩࢻ࣮ࣝࡢࣛࢵࢺ࡬ࡢ 28 ᪥㛫཯᚟ᢞ୚࡟ࡼࡿ⚄⤒⣔࡟ᑐࡍࡿᙳ㡪

41

⥴

⥴ ゝ

➨ 2 ❶࡛ࡣ, ࢢࣜࢩࢻ࣮ࣝࡢẘᛶᶵᗎ࡜ࡋ࡚, ᡂ⇍ື≀ࡢ⚄⤒⣔࡜Ⓨ㐩ᮇࡢື≀ࡢࢽ࣮ࣗ

ࣟࣥ᪂⏕࡟࠾ࡅࡿᗂⱝ࡞᪂⏕⚄⤒ᮎ➃࡟ᑐࡋ࡚, ඹ㏻ࡢᶆⓗᛶࢆᣢࡗࡓ㞀ᐖᛶࡀ♧၀ࡉࢀ

ࡓࠋ

ࢽ࣮ࣗࣟࣥ᪂⏕ࡣᡂ⇍ື≀ࡢᾏ㤿ṑ≧ᅇ SGZ ࡛ࡶࡳࡽࢀ , Ⓨ㐩⚄⤒ẘᛶ≀㉁ࡢᶆⓗ࡜ࡶ

࡞ࡾ࠺ࡿࡇ࡜ࡀ⪃࠼ࡽࢀࡿࠋᢠ⏥≧⭢๣࡜ࡋ࡚▱ࡽࢀࡿ࣓ࢳ࣐ࢰ࣮ࣝࡣ , ࣛࢵࢺⓎ㐩ᮇᭀ㟢

࡟࠾࠸࡚ , ᾏ㤿ṑ≧ᅇ SGZ ࡛ࡢ type-1 ⚄⤒ᖿ⣽⬊࠿ࡽᮍ⇍㢛⢏⣽⬊ࡲ࡛ࡢᗈ࠸⠊ᅖࡢ㢛⢏

⣽⬊⣔㆕ࡢῶᑡཬࡧࡑࢀࢆ཯ᫎࡋࡓṑ≧ᅇ㛛ࡢ GABA ᛶ௓ᅾࢽ࣮ࣗࣟࣥศᕸࡢኚື࡟ࡼࡗ

࡚ࢽ࣮ࣗࣟࣥ᪂⏕㞀ᐖᛶࡀぢฟࡉࢀ࡚࠸ࡿ㸦81㸧 ࠋࡉࡽ࡟ᡂ⇍ࣛࢵࢺ࡬ࡢ࣓ࢳ࣐ࢰ࣮ࣝࡢᢞ

୚࡟ࡼࡗ࡚ࡶ , ྠᵝࡢ GABA ᛶ௓ᅾࢽ࣮ࣗࣟࣥࡢศᕸኚືࡀㄆࡵࡽࢀ࡚࠸ࡿࡇ࡜ࡼࡾ㸦 81 㸧 ,

Ⓨ㐩ᮇཬࡧᡂ⇍ᮇࡢື≀࡟࠾࠸࡚ྠᵝࡢࢽ࣮ࣗࣟࣥ᪂⏕㞀ᐖࢆㄏⓎࡍࡿࡇ࡜ࡀ♧၀ࡉࢀࡿࠋ

ࡑࡢࡓࡵ , SGZ ࡟࠾ࡅࡿ㢛⢏⣽⬊⣔㆕ࢆᣦᶆ࡜ࡋࡓࢽ࣮ࣗࣟࣥ᪂⏕ࡢ᳨⣴ࡸṑ≧ᅇ㛛࡟࠾

ࡅࡿ௓ᅾࢽ࣮ࣗࣟࣥࡢ᳨⣴࡟ࡼࡗ࡚, ᡂ⇍ື≀ࢆ⏝࠸ࡓ▷ᮇ୍⯡ẘᛶヨ㦂࡟࠾࠸࡚Ⓨ㐩ᮇ ཬࡧᡂ⇍ᮇࡢ⚄⤒࡬ࡢᙳ㡪ࢆඹ࡟ホ౯࡛ࡁࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋ

ᮏ❶࡛ࡣ, ᡂ⇍ື≀࡟ᑐࡍࡿ㍈⣴㞀ᐖᛶ≀㉁࡛࠶ࡿࡇ࡜ࡀ᫂ࡽ࠿࡟࡞ࡗࡓࢢࣜࢩࢻ࣮ࣝ

࡟ࡼࡿ , ᡂ⇍ື≀ࡢࢽ࣮ࣗࣟࣥ᪂⏕࡬ࡢᙳ㡪ࢆ☜ㄆࡋ , Ꮚື≀࡛ࡳࡽࢀࡓᙳ㡪࡜ẚ㍑ࡋࡓࠋ ࡇࡢ┠ⓗࡢࡓࡵ࡟ , ࢢࣜࢩࢻ࣮ࣝࡢ 28 ᪥㛫ᙉไ⤒ཱྀᢞ୚ᚋ࡟ , ᾏ㤿ṑ≧ᅇ SGZ ࡢ⣽⬊ቑṪ

࡜࢔࣏ࢺ࣮ࢩࢫ࡞ࡽࡧ࡟㢛⢏⣽⬊⣔㆕ࡢ᳨⣴࡜ṑ≧ᅇ㛛࡟࠾ࡅࡿ௓ᅾࢽ࣮ࣗࣟࣥࡢศᕸࡢ

᳨⣴ࢆ, ච␿⤌⧊໬Ꮫⓗゎᯒ࡟ࡼࡾᐇ᪋ࡋࡓࠋࡉࡽ࡟, ㍈⣴㞀ᐖ≀㉁࡟ࡼࡿᡂ⇍ື≀ࡢ⬻࡬

ࡢᙳ㡪ࢆᤕࡽ࠼ࡿࡓࡵ࡟, ᖏ≧ᅇ, ᾏ㤿ṑ≧ᅇ, ⬻ᱱཬࡧᑠ⬻⓶㉁࡟࠾ࡅࡿ⥙⨶ⓗ㑇ఏᏊⓎ

⌧ࣉࣟࣇ࢓࢖ࣝࡢ⋓ᚓࢆ⾜ࡗࡓࠋࡲࡓ, ⋓ᚓࡋࡓⓎ⌧ࣉࣟࣇ࢓࢖ࣝࢆඖ࡟, Ⓨ⌧ኚືศᏊࡢ

Ⓨ⌧⣽⬊ศᕸࢆච␿⤌⧊໬Ꮫⓗゎᯒࡼࡗ᳨࡚⣴ࡋࡓࠋ

42

ᮦ

ᮦᩱཬࡧ᪉ἲ

໬Ꮫ≀㉁

ࢢࣜࢩࢻ࣮ࣝ㸦CAS No. 556-52-5㸧ࡣ, ࿴ග⣧⸆ᕤᴗᰴᘧ఍♫㸦ᮾி㸧ࡼࡾ㉎ධࡋ, ᐇ㦂

࡟౪ࡋࡓࠋ

౪ヨື≀

4 㐌㱋ࡢ㞝ᛶ Sprague-Dawley ⣔ SPF ࣛࢵࢺࢆ᪥ᮏࢳ࣮ࣕࣝࢫ࣭ࣜࣂ࣮ᰴᘧ఍♫࠿ࡽ㉎ධ ࡋ࡚ᐇ㦂࡟౪ࡋࡓࠋື≀ࡣ ᗘ 23±2°C, ┦ᑐ‵ᗘ 55±15%, ↷᫂ࢧ࢖ࢡࣝ 12 ᫬㛫᫂/12 ᫬㛫 ᬯ᮲௳ࡢ㣫⫱ᐊ࡛, ᗋᩜࢆධࢀࡓࣉࣛࢫࢳࢵࢡ〇ࢣ࣮ࢪ࡟ 3 ࠿ࡽ 4 ༉ࡎࡘ཰ᐜࡋ, ᇶ♏㣫ᩱ

CRF-1 ཬࡧỈ㐨Ỉࢆ⮬⏤࡟ᦤྲྀࡉࡏࡓࠋ

඲࡚ࡢື≀ᐇ㦂ィ⏬ࡣᐇ㦂᪋タ㸦ᮾி㎰ᕤ኱Ꮫ㸧ࡢື≀ᐇ㦂ጤဨ఍ࡢᢎㄆࢆཷࡅ , ඲࡚ࡢ

ື≀ᐇ㦂᧯సࡣࠕື≀ᐇ㦂ࡢ㐺ṇ࡞ᐇ᪋࡟ྥࡅࡓ࢞࢖ࢻࣛ࢖ࣥࠖ 㸦᪥ᮏᏛ⾡఍㆟ , ᖹᡂ 18 ᖺ 6 ᭶ 1 ᪥㸧ࢆ㑂Ᏺࡋࡓࠋ

ᐇ㦂ࢹࢨ࢖ࣥ

NTP ࡟ࡼࡿࢢࣜࢩࢻ࣮ࣝࡢ 13 㐌㛫཯᚟⤒ཱྀᢞ୚ヨ㦂ࡢሗ࿌㸦 59 㸧ࢆཧ⪃࡟ , ㍍ᚤ࡞య㔜 ῶᑡࡀᘬࡁ㉳ࡇࡉࢀࡿ࡜ண᝿ࡉࢀࡿ 200 mg/kg ࢆ㧗⏝㔞࡟タᐃࡋ , 30 mg/kg ࢆప⏝㔞࡟タᐃ ࡋࡓࠋ

1 㐌㛫ࡢ㥆໬㣫⫱ᚋ, 5 㐌㱋ࡢື≀ࢆ 1 ⩌ᙜࡓࡾ 16 ༉ࡎࡘࡢ 3 ⩌࡟↓సⅭ࡟⩌ศࡅࡋ, 0 㸦፹ యᑐ↷⩌㸧, 30 ཬࡧ 200 mg/kg ࡢ⏝㔞࡛ࢢࣜࢩࢻ࣮ࣝࢆ 28 ᪥㛫ᙉไ⤒ཱྀᢞ୚ࡋࡓࠋࢢࣜࢩ ࢻ࣮ࣝࡣ⢭〇Ỉ࡟⁐ゎࡋ࡚ 6 ཬࡧ 40 mg/ml ᾮࢆㄪ〇ࡋ, ᭱᪂ࡢయ㔜ࢆᇶ࡟ 5 ml/kg ࡟࡚ᢞ୚

ࡋࡓࠋ⚄⤒Ꮫⓗ␗ᖖࡢホ౯ࡢࡓࡵ , ື≀ࡢṌ⾜␗ᖖࢆḟࡢ 4 ࡘࡢ࢝ࢸࢦ࣮ࣜ࡟ᇶ࡙ࡁࢫࢥ࢔

௜ࡅࡋࡓࠋࢢ࣮ࣞࢻ 0 㸸ṇᖖṌ⾜ , ࢢ࣮ࣞࢻ 1 㸸㍍ᗘࡢṌ⾜␗ᖖ㸦㍍ᗘࡢᘬࡁࡎࡾṌ⾜㸧 , ࢢ

࣮ࣞࢻ 2 㸸୰➼ᗘࡢṌ⾜␗ᖖ㸦୰➼ᗘࡢᘬࡁࡎࡾṌ⾜㸧 , ࢢ࣮ࣞࢻ 3 㸸㔜ᗘࡢṌ⾜␗ᖖ㸦య

43

㔜ࢆᨭ࠼ࡽࢀࡎ , ᅄ⫥ࡀᗈࡀࡗࡓ≧ែࢆྵࡴ㸧 ࠋホ౯ࡣࣉࣞ࢟ࢩ࢞ࣛࢫ〇࣎ࢵࢡࢫ㸦 90 cm ×

90 cm × 20 cm 㸧ෆ࡛ 3 ศ㛫 , 1 ༉ࡎࡘ┣᳨ୗ࡛ᐇ᪋ࡋࡓࠋᢞ୚ᮇ㛫୰ , య㔜ཬࡧᦤ㣵㔞ࢆ㐌

࡟ 1-2 ᅇ ᐃࡋࡓࠋ᭱⤊ᢞ୚⩣᪥࡟඲౛ࢆ๗᳨ࡋ, 㑇ఏᏊⓎ⌧ゎᯒ, ⑓⌮⤌⧊Ꮫⓗゎᯒ, ච

␿⤌⧊໬Ꮫⓗゎᯒ, ࢔࣏ࢺ࣮ࢩࢫࡢ᳨⣴ཬࡧ⾑Ύ୰࣍ࣝࣔࣥ⃰ᗘ ᐃ࡟౪ࡋࡓࠋ๗᳨᫬, ⬻ 㔜㔞ࢆ ᐃࡋࡓࠋ඲࡚ࡢື≀ࡣ CO

2

/O

2

㯞㓉ୗ࡛⭡኱ື⬦ษ᩿࡟ࡼࡿᨺ⾑⮴Ṛ࡟ࡼࡾᏳᴦṚ ࡉࡏࡓࠋ

⾑ ⾑Ύ୰࣍ࣝࣔࣥ⃰ᗘ ᐃ

ྛ⩌ 10 ౛࡟ࡘ࠸࡚᭱⤊ᢞ୚⩣᪥࡟ CO

₂

/O

₂

㯞㓉ୗ࡛㛤⭡ࡋ, ⭡኱ື⬦࠿ࡽ⾑ᾮࢆ᥇ྲྀࡋࡓࠋ 㐲ᚰศ㞳㸦3,000 r.p.m.×10 ศ㸧࡟ࡼࡾ⾑Ύヨᩱࢆ᥇ྲྀࡋ, -80°C ࡛෾⤖ಖᏑࡋࡓࠋ

thyroid-stimulating hormone 㸦 TSH 㸧 , triiodothyronine 㸦 T

3

㸧ཬࡧ thyroxine 㸦 T

4

㸧ࢆ rodent ELISA test kit 㸦 Endocrine Technologies, Newark, CA, USA 㸧ཬࡧ࣐࢖ࢡࣟࣉ࣮ࣞࢺ࣮ࣜࢲ࣮㸦 FLUOstar Optima, BMG Labtechnologies, Durham, NC, USA 㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋ

㑇ఏᏊⓎ⌧ゎᯒ⏝≉ᐃ⬻㒊఩ࡢ᥇ྲྀ

๗᳨᫬, ྛ⩌ 6 ౛ࡼࡾ᥇ྲྀࡋࡓ඲⬻ࢆ࣓ࢱ࣮࢝ࣥᾮ࡟ᾐₕࡋ, 4°C ࡛ 5 ᫬㛫᣺┞ࡋ࡚ᅛᐃ ࡋࡓࠋࡑࡢᚋ , 99.5% ࢚ࢱࣀ࣮ࣝ࡟࡚ 1 ᫬㛫࠾ࡁ࡟ 3 ᅇ⬺Ỉࡋ , 4°C ୍࡛ᬌ᣺┞ࡋࡓࠋ

኱⬻ࡣ⬻㗪ᆺ㸦ࣈࣞ࢖࣐ࣥࢺࣜࢵࢡࢫ㸸ᐊ⏫ᶵᲔᰴᘧ఍♫ , ᮾி㸧ࢆ⏝࠸࡚ཌࡉ 2 mm ࡢ

ෙ≧᩿ࢫࣛ࢖ࢫࢆస〇ࡋ, ᑠ⬻ࡣཌࡉ 2 mm ࡢ▮≧᩿ࢫࣛ࢖ࢫࢆస〇ࡋࡓࠋ1 mm ᚄࡢ⏕᳨

ࢺࣞࣃࣥ㸦࢝࢖࢖ࣥࢲࢫࢺ࣮ࣜࢬᰴᘧ఍♫, ᒱ㜧㸧ࢆ⏝࠸࡚, ᖏ≧ᅇ, ᑠ⬻⓶㉁, ᾏ㤿ṑ≧ᅇ ཬࡧ⬻ᱱ⤌⧊ࡢ 4 㒊఩ࢆࡃࡾᢤ࠸࡚᥇ྲྀࡋࡓ㸦Fig. 3-1㸧 ࠋ᥇ྲྀࡋࡓࢧࣥࣉࣝࡣ, 99.5%࢚ࢱࣀ

࣮ࣝ࡟ᾐₕࡋ, -80°C ࡟࡚ಖᏑࡋࡓࠋ

࣐࢖ࢡࣟ࢔ࣞ࢖ゎᯒ

࣓ࢱ࣮࢝ࣥᅛᐃ⬻࠿ࡽ᥇ྲྀࡋࡓྛ⬻㒊఩ࡢ⤌⧊ࢧࣥࣉࣝ࠿ࡽ , QIAzol 㸦 Qiagen, Valencia,

44

CA, USA 㸧ཬࡧ RNeasy Mini kit 㸦 Qiagen 㸧ࢆ⏝࠸࡚〇㐀ඖࡢࣉࣟࢺࢥ࣮ࣝ࡟ᚑࡗ࡚ total RNA

ࢆᢳฟࡋ , NanoDrop ND-1000 㸦 Thermo Scientific, Wilmington, DE, USA 㸧ࢆ⏝࠸࡚⃰ᗘࢆ ᐃ ࡋࡓࠋྛ⩌ 3 ౛ࡢྛ⬻㒊఩࡟ࡘ࠸࡚, 61,657 ࣉ࣮ࣟࣈࢆྵࡴ Agilent Whole Rat Genome array Toxplus 60Kx8㸦Agilent Technologies, Inc., Santa Clara, CA, USA㸧ࢆ⏝࠸࡚⥙⨶ⓗ㑇ఏᏊⓎ⌧

ゎᯒࢆ⾜ࡗࡓࠋ ࢧࣥࣉࣝࡢฎ⌮ཬࡧ࣐࢖ࢡࣟ࢔ࣞ࢖ᐇ㦂ࡣ〇㐀ඖࡢࣉࣟࢺࢥ࣮ࣝ࡟ᚑࡗ࡚⾜

ࡗࡓࠋࢥࣥࢺ࣮ࣟࣝ RNA 㸦 Agilent One-Color RNA Spike-In Kit 㸧࡟ྛ⤌⧊ࡼࡾᢳฟࡋࡓ 200 ng ࡢ total RNA ࢆຍ࠼ , Agilent Low Input Quick Amp Labeling 㸦 one-color 㸧ࢆ⏝࠸࡚ Cy3 ࣛ࣋ࣝ

cRNA ࢆస〇ᚋ , RNeasy mini kit 㸦 Qiagen 㸧ࢆ⏝࠸࡚⢭〇 , in situ hybridization Kit 㸦 Agilent

Technologies, Inc.㸧ࢆ⏝࠸࡚᩿∦໬ࡋ, ࣁ࢖ࣈࣜࢲ࢖ࢮ࣮ࢩࣙࣥࢆ⾜ࡗࡓࠋG2505C scanner

㸦Agilent Technologies, Inc.㸧ཬࡧ Agilent Feature Extraction software㸦version 11.7.1.1㸧ࢆ⏝࠸

࡚㑇ఏᏊⓎ⌧ࢹ࣮ࢱࢆྲྀᚓࡋࡓࠋ࣐࢖ࢡࣟ࢔ࣞ࢖ࢹ࣮ࢱࡢゎᯒࡣ Gene Spring GX 㸦 version 11.5.1 㸧 software 㸦 Agilent Technologies, Inc. 㸧ࢆ⏝࠸࡚⾜ࡗࡓࠋ

Real-time RT-PCR ゎ ゎᯒ

0㸦ᑐ↷⩌㸧ཬࡧ 200 mg/kg ⩌ࡢྛ 6 ౛ࡢྛ⬻㒊఩࡟ࡘ࠸࡚, 1 μg ࡢ total RNA ࡼࡾ

SuperScript™ III Reverse Transcriptase㸦Invitrogen Corp., Carlsbad, CA, USA㸧ࢆ⏝࠸࡚

cDNA

ࢆྜᡂࡋࡓࠋ Table 3-1 ࡟♧ࡋࡓ㑇ఏᏊ࡟ࡘ࠸࡚ , mRNA Ⓨ⌧ࡢᐃ㔞ゎᯒࢆᐇ᪋ࡋࡓࠋ PCR ཯ ᛂࡣ SYBR

^®

Green PCR Master Mix 㸦 Applied Biosystems Inc., Foster City, CA, USA 㸧ࢆ⏝࠸ , Prism 7000 Sequence Detection System 㸦Applied Biosystems Inc.㸧࡟࡚, 〇㐀ඖࡢࣉࣟࢺࢥ࣮

ࣝ࡟ᚑࡗ࡚ᐇ᪋ࡋࡓࠋ⏝࠸ࡓࣉࣛ࢖࣐࣮㸦Table 3-1㸧ࡣ, Primer Express software 㸦Version 3.0;

Applied Biosystems Japan Ltd.㸧ࢆ⏝࠸࡚タィࡋࡓࠋྛ㑇ఏᏊࡢ mRNA Ⓨ⌧㔞ࡣᑐ↷⩌࡛ࡢⓎ

⌧್࡟ᑐࡍࡿ┦ᑐ್࡜ࡋ࡚ồࡵ, ෆᅉᛶࢥࣥࢺ࣮ࣟࣝ࡜ࡋ࡚

Hprt

ཬࡧ

Gapdh

㑇ఏᏊࡢ᳨㔞

⥺ࢆồࡵ , 2

^–ΔΔCT

method 㸦 43 㸧࡟࡚⟬ฟࡋࡓࠋ

⑓⌮⤌⧊Ꮫⓗゎᯒ

45

๗᳨᫬ , ྛ⩌ 10 ༉ࡼࡾ⬻ , ୕ཫ⚄⤒ , ᆘ㦵⚄⤒ཬࡧ⬨㧊ࢆ᥇ྲྀࡋ , ⑓⌮⤌⧊Ꮫⓗゎᯒ࡟౪ ࡋࡓࠋ ᆘ㦵⚄⤒ࡣ , ᥇ྲྀ๓ 3 ศ㛫ࣈ࢔ࣥᾮࢆᾐₕࡉࡏ࡚ in situ ᅛᐃࢆ⾜ࡗࡓࠋ⬨㧊ࡣ㢁㒊 , ⬚ 㒊ཬࡧ⭜㒊࡟ࡘ࠸࡚, ᳝㦵࡜୍ᣓࡋ࡚᥇ྲྀࡋࡓࠋ᥇ྲྀࡋࡓ⤌⧊ࡣࣈ࢔ࣥᾮ࡟ᾐₕࡋ, 4°C ࡛

୍ᬌ᣺┞ࡋ࡚ᅛᐃࡋ, ๭㠃ࢆస〇ࡋࡓࠋ⬨㧊ࡣ, ᅛᐃᚋࣉࣛࣥ-ࢡࣜࣗࢡࣟᾮ࡟ᾐₕࡋ, ᐊ 

࡛ 48 ᫬㛫᣺┞ࡋ࡚⬺⅊ࡋࡓᚋ, 5%◲㓟ࢼࢺ࣒ࣜ࢘ᾮ࡟ᾐₕࡋ, ᐊ ࡛ 48 ᫬㛫᣺┞ࡋ࡚୰࿴

ࡋࡓࠋ኱⬻ࡣ Bregma ࡢᚋ᪉⣙ -3.5 mm ࡢ 1 ࢝ᡤ࡛ෙ≧᩿๭㠃ࢆస〇ࡋ , ᑠ⬻ཬࡧᘏ㧊ࡣ▮

≧᩿๭㠃ࢆస〇ࡋࡓࠋ⤌⧊ࡣࣃࣛࣇ࢕ࣥໟᇙࡋ

, 3μm

ཌࡢษ∦ࢆస〇ࡋࡓࠋษ∦ࡣ࣐࣊ࢺ࢟

ࢩ࣭࢚ࣜࣥ࢜ࢪࣥᰁⰍࢆ⾜ࡗࡓࠋ

ච ච␿⤌⧊໬Ꮫⓗゎᯒཬࡧ࢔࣏ࢺ࣮ࢩࢫࡢ᳨⣴

ྛ⩌ 10 ༉ࡢ⬻⤌⧊ษ∦࡟ࡘ࠸࡚ , Table 3-2 ࡟♧ࡋࡓᢠయࢆ⏝࠸࡚ච␿⤌⧊໬ᏛⓗᰁⰍࢆ

⾜ࡗࡓࠋࡉࡽ࡟ , ྛ⩌ 10 ༉ࡢ୕ཫ⚄⤒⤌⧊ษ∦࡟ࡘ࠸࡚ , ᢠ p-NF-H ᢠయ㸦 Table 3-2 㸧ࢆ⏝

࠸࡚ච␿⤌⧊໬ᏛⓗᰁⰍࢆ⾜ࡗࡓࠋྛ୍ḟᢠయࡣ୍ᬌ㸦 4°C㸧཯ᛂࡉࡏ ,

3,3’-diaminobenzidine/H₂

O

₂

ⓎⰍࢆ⏝࠸࡚ ABC ἲ㸦Vector Laboratories Inc., Burlingame, CA,

U.S.A.㸧࡟ࡼࡿච␿ᰁⰍࢆ⾜ࡗࡓࠋ࡞࠾, ࢡ࢚ࣥ㓟⦆⾪ᾮ㸦10 mM, pH 6.0㸧ࢆ⏝࠸࡚, Tbr2

ཬࡧ activity-regulated cytoskeleton-associated protein㸦Arc㸧࡟ࡘ࠸࡚ࡣ࣮࢜ࢺࢡ࣮ࣞࣈฎ⌮

㸦 121 °C, 10 ศ㸧 , Calb1, Calb2 ཬࡧ Pvalb ࡟ࡘ࠸࡚ࡣ࣐࢖ࢡ࢙࣮ࣟ࢘ࣈฎ⌮㸦 90 °C, 10 ศ㸧

࡟ࡼࡿᢠཎࡢ㈿ά໬ฎ⌮ࢆ⾜ࡗࡓࠋ

ࡲࡓ, ᢠ DCX ᢠయ࡜ᢠ TUC-4 ᢠయ࡟ࡼࡿ஧㔜ᰁⰍࢆᐇ᪋ࡋࡓࠋTUC-4 ࡣ

3,3’-diaminobenzidine/H2

O

2

ⓎⰍࢆ⏝࠸࡚ ABC ἲ㸦Elite ABC kit, Vector Laboratories Inc.㸧࡟ࡼ

ࡾ, DCX ࡣ Vector Red Alkaline Phosphate Substrate Kit I㸦Vector Laboratories Inc.㸧ࡢⓎⰍࢆ⏝

࠸࡚ ABC ἲ㸦ABC-AP kit, Vector Laboratories Inc.㸧࡟ࡼࡾ⾜ࡗࡓࠋ

ᾏ㤿ṑ≧ᅇࡢ SGZ ࡟࠾ࡅࡿ࢔࣏ࢺ࣮ࢩࢫࡢホ౯࡟ࡘ࠸࡚ࡣ , cleaved caspase-3 ᢠయࢆ⏝࠸

ࡓච␿⤌⧊Ꮫⓗゎᯒ࡟ຍ࠼࡚ , TUNEL ἲ㸦 Apop Tag

^®

in situ apoptosis detection kit, Millipore

Corporation 㸧ࢆ⏝࠸࡚〇㐀ඖࡢࣉࣟࢺࢥ࣮ࣝ࡟ᚑࡗ᳨࡚ฟࢆ⾜ࡗࡓࠋ

46 ᶆᮏࡣ࣐࣊ࢺ࢟ࢩࣜࣥ࡟ࡼࡾᑐẚᰁⰍࢆ᪋ࡋࡓࠋ

ᾏ㤿ṑ≧ᅇ SGZ ࡟࠾࠸࡚ , 㢛⢏⣽⬊ࡢ⣔㆕࡛࠶ࡿ TUC-4, DCX, Tbr2, Pax6 ཬࡧ GFAP 㝧ᛶ

⣽⬊ᩘ, ࢔࣏ࢺ࣮ࢩࢫࡢᣦᶆ࡛࠶ࡿ cleaved caspase-3 ཬࡧ TUNEL 㝧ᛶ⣽⬊ᩘ, ቑṪ⣽⬊ࡢᣦ ᶆ࡛࠶ࡿ PCNA 㝧ᛶ⣽⬊ᩘࡢ᳨⣴ࢆᕥྑ࡟ࡘ࠸࡚⾜࠸, SGZ ࡢ㛗ࡉࢆ⏝࠸࡚ࣀ࣮࣐ࣛ࢖ࢬࡋ ࡓ㸦Fig. 3-2㸧 ࠋᾏ㤿ṑ≧ᅇࡢ㢛⢏⣽⬊ᒙ࡟࠾࠸࡚, NeuN 㝧ᛶ⣽⬊ᩘࡢ᳨⣴ࢆᕥྑ࡟ࡘ࠸࡚⾜

࠸ , SGZ ࡢ㛗ࡉ࠶ࡿ࠸ࡣ㢛⢏⣽⬊ᒙࡢ㠃✚ࢆ⏝࠸࡚ࣀ࣮࣐ࣛ࢖ࢬࡋࡓ㸦 Fig. 3-2 㸧 ࠋᾏ㤿ṑ≧

ᅇ㛛࡟࠾࠸࡚ , GABA ᛶ௓ᅾࢽ࣮ࣗࣟࣥ࡟Ⓨ⌧ࡍࡿศᏊࡢ reelin, Calb1, Calb2 ཬࡧ Pvalb ࡞ࡽ

ࡧ࡟ᡂ⇍ࢽ࣮ࣗࣟࣥࡢᣦᶆ࡛࠶ࡿ NeuN ࡟ࡘ࠸࡚㝧ᛶ⣽⬊ᩘࡢ᳨⣴ࢆᕥྑ࡟ࡘ࠸࡚⾜࠸ , ṑ

≧ᅇ㛛ࡢ㠃✚ࢆ⏝࠸࡚ࣀ࣮࣐ࣛ࢖ࢬࡋࡓ㸦Fig. 3-2㸧 ࠋᾏ㤿ṑ≧ᅇࡢ㢛⢏⣽⬊ᒙ࡟࠾࠸࡚, Arc, FBJ osteosarcoma oncogene㸦Fos㸧ཬࡧ jun proto-oncogene㸦Jun㸧㝧ᛶ⣽⬊ᩘࡢ᳨⣴ࢆᕥྑ࡟

ࡘ࠸࡚⾜࠸ , 㢛⢏⣽⬊ᒙࡢ㛗ࡉࢆ⏝࠸࡚ࣀ࣮࣐ࣛ࢖ࢬࡋࡓ㸦 Fig. 3-3 㸧 ࠋࡲࡓ , ᖏ≧ᅇ࡟ศᕸ ࡍࡿ Arc 㝧ᛶ⣽⬊ᩘࢆ , ⬻ᱱཬࡧᖏ≧᮰࡛ᅖࡲࢀࡓ㡿ᇦ᳨࡛⣴ࡋ , 㠃✚࡛ࣀ࣮࣐ࣛ࢖ࢬࡋࡓ 㸦 Fig. 3-4 㸧 ࠋᑠ⬻㢛⢏⣽⬊ᒙ࡟ศᕸࡍࡿ Fos 㝧ᛶ⣽⬊ᩘࢆ , 10

^th

folium ᳨࡛⣴ࡋ , 㠃✚࡛ࣀ࣮

࣐ࣛ࢖ࢬࡋࡓ㸦Fig. 3-5㸧 ࠋ

ᾏ㤿ṑ≧ᅇࡢ㢛⢏⣽⬊ᒙ࡟࠾ࡅࡿ Arc, Fos ཬࡧ Jun 㝧ᛶ⣽⬊ᩘ, ᖏ≧ᅇ࡟ศᕸࡍࡿ Arc 㝧 ᛶ⣽⬊ᩘཬࡧᑠ⬻㢛⢏⣽⬊ᒙ࡟ศᕸࡍࡿ Fos 㝧ᛶ⣽⬊ᩘࡣ, ➨ 2 ❶࡟࡚⾜ࡗࡓⓎ㐩ᮇᭀ㟢ᐇ 㦂࡟࡚ᚓࡽࢀࡓ⏕ᚋ 21 ཬࡧ 77 ᪥ࡢ㞝Ꮚື≀ࡢ⬻ษ∦ࢆ⏝࠸࡚ࡶྠᵝ࡟ච␿ᰁⰍࢆ⾜࠸ , ゎ ᯒࢆ⾜ࡗࡓࠋ

⤫ ⤫ィゎᯒ

య㔜, ᦤ㣵㔞, ⬻㔜㔞, ච␿⤌⧊໬Ꮫⓗゎᯒཬࡧ࣍ࣝࣔࣥ ᐃࡢࢹ࣮ࢱࡣ, Bartlett ἲ࡟ࡼ

ࡾྛ⩌ࡢ➼ศᩓᛶࡢ᳨ᐃࢆ⾜࠸, ➼ศᩓࡢሙྜࡣ Dunnett ἲ, ୙➼ศᩓࡢሙྜࡣ Steel ἲࢆ⏝

࠸࡚ᑐ↷⩌࡜ྛᢞ୚⩌ࡢᕪࢆ᳨ᐃࡋࡓࠋ real-time RT-PCR ゎᯒཬࡧච␿⤌⧊໬Ꮫⓗゎᯒ࡟࠾

ࡅࡿ 2 ⩌㛫ࡢẚ㍑ࡣ , F ᳨ᐃ࡟ࡼࡾྛ⩌ࡢ➼ศᩓᛶࡢ᳨ᐃࢆ⾜࠸ , ➼ศᩓࡢሙྜࡣ Student ࡢ

t ᳨ᐃ , ୙➼ศᩓࡢሙྜࡣ Aspin-Welch ࡢ t ᳨ᐃࢆ⏝࠸࡚ᑐ↷⩌࡜ᢞ୚⩌ࡢᕪࢆ᳨ᐃࡋࡓࠋ࣐

47

࢖ࢡࣟ࢔ࣞ࢖ゎᯒࡢࢹ࣮ࢱࡣ Welch ࡢ t ᳨ᐃࢆ⏝࠸࡚ᑐ↷⩌࡜ᢞ୚⩌ࡢᕪࢆ᳨ᐃࡋࡓࠋṌ⾜

␗ᖖࡢࢫࢥ࢔ཬࡧ⑓⌮⤌⧊Ꮫⓗゎᯒ࡟࠾ࡅࡿ⑓ኚࡢ⛬ᗘࡣ , Mann-Whitney ࡢ U ᳨ᐃ࡟ࡼࡾ

᳨ᐃࢆ⾜ࡗࡓࠋ

48

⤖

⤖ ᯝ

୍⯡≧ែ, య㔜, ᦤ㣵㔞ཬࡧ⬻㔜㔞

200 mg/kg ⩌࡛, ᢞ୚➨ 18 ᪥࠿ࡽ๗᳨᪥ࡲ࡛Ṍ⾜␗ᖖ㸦ࡦࡁࡎࡾṌ⾜㸧ࡀࡳࡽࢀࡓࠋᢞ୚

➨ 21 ᪥௨㝆, ᑐ↷⩌࡜ẚ㍑ࡋ࡚᭷ព࡞ᕪࡀㄆࡵࡽࢀ, ๗᳨᪥࡟ࡣ 200 mg/kg ⩌ࡢ඲౛࡛ࡳࡽ

ࢀࡓࠋ 0, 30 mg/kg ⩌࡛ࡣ␗ᖖࡣㄆࡵࡽࢀ࡞࠿ࡗࡓ㸦 Table 3-3 㸧 ࠋయ㔜ࡣ , 200 mg/kg ⩌࡛ᢞ୚

➨ 3 ᪥࠿ࡽᢞ୚➨ 28 ᪥ࡲ࡛ , ᑐ↷⩌࡜ẚ㍑ࡋ࡚᭷ព࡞ప್ࢆ♧ࡋࡓ㸦 Table 3-4 㸧 ࠋᦤ㣵㔞ࡣ ,

200 mg/kg ⩌࡛ᢞ୚➨ 3 ᪥࠿ࡽᢞ୚➨ 28 ᪥ࡲ࡛ , ᑐ↷⩌࡜ẚ㍑ࡋ࡚᭷ព࡞ప್ࢆ♧ࡋࡓ

㸦Table 3-4㸧 ࠋ⬻㔜㔞ࡣ, 200 mg/kg ⩌࡛⤯ᑐ㔜㔞ࡢ᭷ព࡞ప್ཬࡧ┦ᑐ㔜㔞ࡢ᭷ព࡞㧗್ࡀ ㄆࡵࡽࢀࡓ㸦Table 3-5㸧 ࠋ

⾑Ύ୰⏥≧⭢㛵㐃࣍ࣝࣔࣥ⃰ᗘ

TSH ཬࡧ T

3

ࡢ᭷ព࡞㧗್ࡀ 200 mg/kg ⩌࡛ㄆࡵࡽࢀࡓ㸦 Table 3-6 㸧 ࠋ T

4

ࡢ᭷ព࡞㧗್ࡀ 30

mg/kg ⩌࡛ㄆࡵࡽࢀࡓࡀ, 200 mg/kg ⩌࡜ᑐ↷⩌࡜ࡢ㛫࡟ᕪࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦Table 3-6㸧 ࠋ

࣐࢖ࢡࣟ࢔ࣞ࢖ゎᯒ

࣐࢖ࢡࣟ࢔ࣞ࢖ゎᯒ⤖ᯝࢆ Table 3-7 㹼 3-14 ࡟♧ࡋࡓࠋ

ᾏ㤿ṑ≧ᅇ࡛ࡣ , ࢽ࣮ࣗࣟࣥ᪂⏕ , ㍈⣴ᙧᡂཬࡧ⚄⤒✺㉳ᙧᡂ࡟㛵ࢃࡿ㑇ఏᏊ⩌ࡢⓎ⌧㔞 ኚື㸦ῶᑡ㸸Bmp4, Bdnf, Kdr ཬࡧ

Atf3,

ቑຍ㸸Runx3 ཬࡧ

Cxcl12㸧ཬࡧࢩࢼࣉࢫఏ㐩࡟㛵ࢃ

ࡿ㑇ఏᏊࡢኚື㸦ῶᑡ㸸Crh㸧ࡀ 200 mg/kg ⩌࡛ㄆࡵࡽࢀࡓࠋ

⬻ᱱ࡛ࡣ, ࢢࣜ࢔⣽⬊ࡢᡂ⇍ཬࡧᶵ⬟࡟㛵ࢃࡿ㑇ఏᏊ⩌ࡢⓎ⌧㔞ኚື㸦ῶᑡ㸸Cntf ཬࡧ

Tnc,

ቑຍ㸸Ptprc㸧ࡀ 200 mg/kg ⩌࡛ㄆࡵࡽࢀࡓࠋ

ᖏ≧ᅇ࡛ࡣ , ㍈⣴ᙧᡂཬࡧ⚄⤒✺㉳ᙧᡂ࡟㛵ࢃࡿ㑇ఏᏊ⩌ࡢⓎ⌧㔞ኚື㸦

Nr4a3, Btg2, Plxnd1, Slitrk2, Rasgrf1, Bhlhe22, Atf3, Klf4, Cyr61

ཬࡧ

Sema5a

㸧ཬࡧࢩࢼࣉࢫఏ㐩࡟㛵ࢃࡿ㑇

ఏᏊ⩌ࡢⓎ⌧㔞ኚື㸦

Cartpt, Slc17a6, Arc

ཬࡧ

Grik3

㸧ࡀ 200 mg/kg ⩌࡛ㄆࡵࡽࢀ , ࡯࡜ࢇ࡝

49

࡛ῶᑡࡋࡓࠋࡇࢀࡽࡢ࠺ࡕ

Cyr61

ཬࡧ

Btg2

ࡢⓎ⌧㔞ῶᑡࡣ 30 mg/kg ⩌࡛ࡶࡳࡽࢀࡓࠋ ᑠ⬻⓶㉁࡛ࡣ , ࢽ࣮ࣗࣟࣥ᪂⏕ , ㍈⣴ᙧᡂཬࡧ⚄⤒✺㉳ᙧᡂ࡟㛵ࢃࡿ㑇ఏᏊ⩌ࡢⓎ⌧㔞ኚ

ື㸦Bmp4, Nr4a3, Tiam1, Nrep, Ntf3 ཬࡧ

Grik3㸧ཬࡧࢩࢼࣉࢫఏ㐩࡟㛵ࢃࡿ㑇ఏᏊ⩌ࡢⓎ⌧㔞

ኚື㸦Ptgs2, Rims3 ཬࡧ

Camk4㸧ࡀ

200 mg/kg ⩌࡛ㄆࡵࡽࢀ, ࡯࡜ࢇ࡝࡛ῶᑡࡋࡓࠋ

Real-time RT-PCR ゎ ゎᯒ

࣐࢖ࢡࣟ࢔ࣞ࢖ゎᯒ࡟ࡼࡿ㑇ఏᏊⓎ⌧ࣉࣟࣇ࢓࢖ࣝࢆ᳨ドࡍࡿࡓࡵ , ྛ⬻㒊఩࡟࠾࠸࡚

200 mg/kg ⩌࡛Ⓨ⌧ኚືࡀࡳࡽࢀࡓ㑇ఏᏊࡢ୰࠿ࡽࢽ࣮ࣗࣟࣥࡢⓎ㐩ࡸྍረᛶ࡟㛵㐃ࡍࡿ

ࡶࡢࢆ࠸ࡃࡘ࠿㑅ᢥࡋ, real-time RT-PCR ࡟ࡼࡿ㑇ఏᏊⓎ⌧ゎᯒࢆᐇ᪋ࡋࡓ࡜ࡇࢁ, ࠸ࡎࢀ

ࡢ㑇ఏᏊࡶ࣐࢖ࢡࣟ࢔ࣞ࢖ࢹ࣮ࢱ࡜ྠᵝࡢ᭷ព࡞ኚືࢆ♧ࡋࡓ㸦Table 3-15㸧 ࠋ

ࡲࡓ , ᾏ㤿ṑ≧ᅇࡢࢽ࣮ࣗࣟࣥྍረᛶཬࡧ㧊㠧ᙧᡂ㛵㐃㑇ఏᏊࡢⓎ⌧ゎᯒ࡛ࡣ , Mag,

Ncam2

ཬࡧ

Mal

ࡢⓎ⌧㔞ࡢ᭷ព࡞ቑຍࡀ 200 mg/kg ⩌࡛ㄆࡵࡽࢀࡓ㸦 Table 3-16 㸧 ࠋ࢔࣏ࢺ࣮

ࢩࢫ , ⣽⬊ቑṪཬࡧ㢛⢏⣽⬊ࡢศ໬⣔㆕࣐࣮࣮࢝࡟࠾࠸࡚ࡣ , ࠸ࡎࢀࡶⓎ⌧㔞ࡢኚືࡣࡳࡽ

ࢀ࡞࠿ࡗࡓ㸦Table 3-16㸧 ࠋ

⑓⌮⤌⧊Ꮫⓗ᳨ᰝ

200 mg/kg ⩌࡟࠾࠸࡚ , ୕ཫ⚄⤒ࡢ⚄⤒⠇⣽⬊࡛୰ᚰᛶⰍ㉁⼥ゎࡀ , ᆘ㦵⚄⤒࡛㍈⣴ኚᛶ

ࡀࡳࡽࢀ㸦 Fig. 3-6 㸧 , ࠸ࡎࢀࡶ⑓ኚࡢ⛬ᗘࡣᑐ↷⩌࡜ẚ㍑ࡋ࡚⤫ィᏛⓗ࡟᭷ព࡛࠶ࡗࡓ 㸦Table 3-17㸧 ࠋ30 mg/kg ⩌࡛ࡣࡇࢀࡽࡢኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓࠋ

200 mg/kg ⩌ࡢ඲౛࡟, ᑠ⬻ࡢ㢛⢏⣽⬊ᒙཬࡧᘏ㧊⫼⣴࡛ NF-L 㝧ᛶࡢࢫࣇ࢙ࣟ࢖ࢻᙧᡂ

ࡀㄆࡵࡽࢀࡓ㸦Fig. 3-7㸧 ࠋ200 mg/kg ⩌࡛ p-NF-H 㝧ᛶ⚄⤒⠇⣽⬊ࡢ᭷ព࡞ῶᑡࡀㄆࡵࡽࢀ

ࡓ㸦Fig. 3-8㸧 ࠋ30 mg/kg ⩌࡛ࡣࢫࣇ࢙ࣟ࢖ࢻᙧᡂཬࡧ p-NF-H 㝧ᛶ⚄⤒⠇⣽⬊ࡢኚ໬ࡣㄆࡵ

ࡽࢀ࡞࠿ࡗࡓࠋ

SGZ ࡟࠾ࡅࡿච␿⤌⧊໬Ꮫⓗゎᯒ࡟ࡼࡿ㢛⢏⣽⬊ࡢศ໬⣔㆕ࡢ᳨⣴

50

200 mg/kg ⩌࡛ DCX ཬࡧ TUC-4 㝧ᛶ⣽⬊ᩘࡢ᭷ព࡞ῶᑡࡀㄆࡵࡽࢀࡓࠋ GFAP, Pax6 ཬࡧ

Tbr2 㝧ᛶ⣽⬊ᩘ࡟ኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦 Fig. 3-9 㸧ࠋ

ࡉࡽ࡟, ࡇࢀࡽࡢኚືࢆࡼࡾヲ⣽࡟᳨⣴ࡍࡿࡓࡵ࡟ᐇ᪋ࡋࡓ TUC-4 ࡜ DCX ࡢ஧㔜ᰁⰍ࡛

ࡣ, 200 mg/kg ⩌࡛ TUC-4 㝧ᛶ/DCX 㝧ᛶ⣽⬊ᩘཬࡧ TUC-4 㝜ᛶ/DCX 㝧ᛶ⣽⬊ᩘࡢ᭷ព࡞ῶ ᑡࡀㄆࡵࡽࢀࡓࡀ, TUC-4 㝧ᛶ/DCX 㝜ᛶ⣽⬊ᩘ࡟ኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦Fig. 3-10㸧 ࠋ

ᾏ ᾏ㤿ṑ≧ᅇ㛛࡟࠾ࡅࡿච␿⤌⧊໬Ꮫⓗゎᯒ࡟ࡼࡿࢽ࣮ࣗࣟࣥศᕸࡢ᳨⣴

200 mg/kg ⩌࡛ NeuN, reelin ཬࡧ Calb2 㝧ᛶ⣽⬊ᩘࡢ᭷ព࡞ቑຍࡀㄆࡵࡽࢀࡓࠋ Calb1 ཬࡧ

Pvalb 㝧ᛶ⣽⬊ᩘ࡟ኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦Fig. 3-11㸧 ࠋ

SGZ ࡟࠾ࡅࡿ࢔࣏ࢺ࣮ࢩࢫ, ⣽⬊ቑṪཬࡧᾏ㤿㢛⢏⣽⬊ᒙ࡟࠾ࡅࡿᡂ⇍ࢽ࣮ࣗࣟࣥࡢ

᳨⣴

SGZ ࡢ TUNEL 㝧ᛶ࢔࣏ࢺ࣮ࢩࢫ⣽⬊ , cleaved caspase-3 㝧ᛶ⣽⬊ཬࡧ PCNA 㝧ᛶ⣽⬊ᩘ

࡟ᑐ↷⩌࡜ᢞ୚⩌ࡢ㛫࡟ᕪࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦Fig. 3-12㸧 ࠋࡲࡓ, 㢛⢏⣽⬊ᒙࡢ NeuN 㝧ᛶᡂ

⇍ࢽ࣮ࣗࣟࣥᩘ࡟ᑐ↷⩌࡜ 200 mg/kg ⩌ࡢ㛫࡛ᕪࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦Table 3-18㸧 ࠋ

ᾏ㤿㢛⢏⣽⬊ᒙ, ᖏ≧ᅇཬࡧᑠ⬻⓶㉁࡟࠾ࡅࡿ Arc, Fos ཬࡧ Jun 㝧ᛶࢽ࣮ࣗࣟࣥࡢ᳨⣴

ᾏ㤿ṑ≧ᅇࡢ㢛⢏⣽⬊ᒙ୰ࡢ Arc, Fos ཬࡧ Jun 㝧ᛶ⣽⬊ᩘࡢ᭷ព࡞ῶᑡࡀ 200 mg/kg ⩌࡛

ㄆࡵࡽࢀࡓ 㸦Fig. 3-13㹼3-15㸧 ࠋ Fos 㝧ᛶ⣽⬊ᩘࡢῶᑡࡣ 30 mg/kg ⩌࡛ࡶࡳࡽࢀࡓ 㸦Fig. 3-14㸧 ࠋ ᖏ≧ᅇࡢ Arc 㝧ᛶ⣽⬊ᩘࡢ᭷ព࡞ῶᑡࡀ 200 mg/kg ⩌࡛ㄆࡵࡽࢀࡓ㸦Fig. 3-13㸧 ࠋࡲࡓ,ᑠ⬻

㢛⢏⣽⬊ᒙ୰ࡢ Fos 㝧ᛶ⣽⬊ᩘࡢ᭷ព࡞ῶᑡࡀ 30 ཬࡧ 200 mg/kg ⩌࡛ㄆࡵࡽࢀࡓ 㸦Fig. 3-14㸧 ࠋ

➨ 2 ❶࡛⾜ࡗࡓⓎ㐩ᮇᭀ㟢ᐇ㦂࡟࠾ࡅࡿゎᯒ࡛ᚓࡽࢀࡓ㞝Ꮚື≀ࡢ⬻࡛ࡣ, ⏕ᚋ 21 ᪥࡟ࡣ

࠸ࡎࢀࡢ⬻㒊఩࡛ࡶ Arc, Fos ཬࡧ Jun 㝧ᛶ⣽⬊ᩘ࡟ኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓࠋ⏕ᚋ 77 ᪥࡟ࡣ ᾏ㤿ṑ≧ᅇࡢ㢛⢏⣽⬊ᒙ୰ࡢ Arc 㝧ᛶ⣽⬊ᩘࡢ᭷ព࡞ቑຍࡀ 1,000 ppm ⩌࡛ , Fos 㝧ᛶ⣽⬊ᩘ

ࡢ᭷ព࡞ቑຍࡀ 300 ཬࡧ 1,000 ppm ⩌࡛ㄆࡵࡽࢀࡓ㸦 Fig. 3-13, 3-14 㸧 ࠋᖏ≧ᅇཬࡧᑠ⬻⓶㉁

51

࡛ࡣኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓࠋ

52

⪃

⪃ ᐹ

ࢢࣜࢩࢻ࣮ࣝࡢ 28 ᪥㛫཯᚟ᢞ୚ࡢ⤖ᯝ, ㍈⣴㞀ᐖࢆㄏⓎࡋࡓ 200 mg/kg ⩌࡛, ᾏ㤿ṑ≧ᅇ

SGZ ࡢ DCX ࡲࡓࡣ TUC-4 㝧ᛶ⣽⬊ࡢῶᑡࢆᘬࡁ㉳ࡇࡋࡓࠋDCX ࡣ Type-2b ๓㥑⣽⬊࠿ࡽ

ᮍ⇍㢛⢏⣽⬊ࡢ⠊ᅖࡢ⣽⬊࡛Ⓨ⌧ࡍࡿ࣐࣮࣮࢝ศᏊ࡛࠶ࡿࡀ㸦34, 35㸧, Type-2b ๓㥑⣽⬊࡛

Ⓨ⌧ࡍࡿ Tbr2 㝧ᛶ⣽⬊㸦 26 㸧ᩘ࡟ኚືࡀ࡞࠸ࡇ࡜࠿ࡽ , Type-2b ๓㥑⣽⬊࡬ࡢᙳ㡪ࡣ࡞࠸ࡶ

ࡢ࡜ุ᩿ࡉࢀࡓࠋࡲࡓ , ஧㔜ᰁⰍ࡟ࡼࡗ࡚☜ㄆࡉࢀࡓ DCX 㝧ᛶ /TUC-4 㝧ᛶཬࡧ DCX 㝧ᛶ

/TUC-4 㝜ᛶ⣽⬊ࡢῶᑡ࠿ࡽࡶ Type-3 ๓㥑⣽⬊ཬࡧᮍ⇍㢛⢏⣽⬊࡟ᑐࡍࡿᙳ㡪࡛࠶ࡿ࡜♧၀

ࡉࢀࡓࠋࡋ࠿ࡋ, NeuN 㝧ᛶᡂ⇍㢛⢏⣽⬊ᩘ࡟ᙳ㡪ࢆཬࡰࡉ࡞࠸ࡇ࡜ࡀ☜ㄆࡉࢀ, ᮏᐇ㦂࡟࠾

ࡅࡿᢞ୚ᮇ㛫࡛ࡣ, Type-3 ๓㥑⣽⬊࠿ࡽᮍ⇍㢛⢏⣽⬊ᩘࡢῶᑡࡣ, ᡂ⇍㢛⢏⣽⬊ࡢῶᑡ࡟ࡲ

࡛ᙳ㡪ࡋ࡞࠸ࡇ࡜ࡀ♧၀ࡉࢀࡓࠋ

➨ 2 ❶࡟࠾ࡅࡿⓎ㐩ᮇᭀ㟢࡛ࡣ , ᾏ㤿ṑ≧ᅇ SGZ ࡢ TUC-4 㝧ᛶ⣽⬊ࡢῶᑡࡢࡳࢆぢฟࡋ , ࢢࣜࢩࢻ࣮ࣝࡀⓎ㐩⚄⤒ẘ࡜ࡋ࡚ᗂⱝ࡞⚄⤒ᮎ➃࡬ࡢᶆⓗᛶ࡟ࡼࡗ࡚ᮍ⇍㢛⢏⣽⬊ࢆ┤᥋ യᐖࡍࡿࡇ࡜ࡀ᥎ ࡉࢀࡓࠋᮏ❶࡟࠾ࡅࡿ 28 ᪥㛫ᢞ୚࡛ࡣ, ᢞ୚㔞ࡸᢞ୚ᮇ㛫ࡢ㐪࠸ࡀᙳ 㡪ࡍࡿ࡜⪃࠼ࡽࢀࡿࡀ, DCX 㝧ᛶ⣽⬊ࡢῶᑡࡶ☜ㄆࡉࢀࡓࡇ࡜࠿ࡽⓎ㐩ᮇᭀ㟢࡜ẚ㍑ࡋ࡚

ࡼࡾᙉ࠸ࢽ࣮ࣗࣟࣥ᪂⏕㞀ᐖࡀ♧၀ࡉࢀࡓࠋDCX ࡣᚤᑠ⟶ࢆᏳᐃ໬ࡋ, ⚄⤒✺㉳ఙ㛗ࡸࢽ

࣮ࣗࣟࣥࡢ⛣ືࢆไᚚࡍࡿࡇ࡜࠿ࡽ㸦 12, 30 㸧 , DCX Ⓨ⌧⣽⬊ࡣ᪂⏕⚄⤒ᮎ➃࡬ࡢᶆⓗᛶࢆ

♧ࡍ≀㉁ࡢᙳ㡪ࢆཷࡅࡿࡇ࡜ࡀ⪃࠼ࡽࢀࡿࠋ ⑓⌮Ꮫⓗゎᯒཬࡧච␿⤌⧊໬Ꮫⓗゎᯒ࡟࠾ࡅࡿ

ᙧែⓗホ౯࡟ࡼࡗ࡚ᾏ㤿ṑ≧ᅇ࡟࠾ࡅࡿ㍈⣴ኚᛶࡣ☜ㄆ࡛ࡁ࡞࠿ࡗࡓࡀ, Ⓨ㐩ᮇᭀ㟢᫬࡟

☜ㄆࡉࢀࡓ㍈⣴ᙧᡂࡸ㧊㠧ᙧᡂ࡟㛵୚ࡍࡿ

Mag, Ncam2

ཬࡧ

Mal

➼ࡢ㑇ఏᏊⓎ⌧㔞ࡢῶᑡࡣ 28 ᪥㛫ᢞ୚ᚋࡢᾏ㤿ṑ≧ᅇ࡛ࡶ☜ㄆࡉࢀ, SGZ ࡟࠾ࡅࡿࢽ࣮ࣗࣟࣥ᪂⏕ࡢᚋᮇ࡟⏕ࡌࡿ⚄

⤒✺㉳ᙧᡂ࡟㛵㐃ࡋࡓ⚄⤒ྍረᛶ࡬ࡢᙳ㡪࡜ࡋ᳨࡚ฟࡉࢀࡓࡶࡢ࡜♧၀ࡉࢀࡓࠋ ௨ୖࡢࡇ࡜

࠿ࡽ , ࢢࣜࢩࢻ࣮ࣝࡣᡂ⇍ື≀࡛࠶ࡗ࡚ࡶ , Ⓨ㐩ᮇᭀ㟢᫬࡜ྠᵝ࡟⚄⤒✺㉳ᙧᡂ㐣⛬ࢆྵࡴ

ࢽ࣮ࣗࣟࣥ᪂⏕ࡢᚋᮇ࡛ࡢ␗ᖖࢆᘬࡁ㉳ࡇࡍࡇ࡜ࡀ♧ࡉࢀ , ࡍ࡞ࢃࡕᡂ⇍ࢽ࣮ࣗࣟࣥࡢ㍈

⣴ᮎ➃ࢆᶆⓗ࡜ࡍࡿ⚄⤒ẘࡀ , ྠᵝࡢ⣽⬊ᶆⓗᶵᗎ࡟ࡼࡗ࡚ᡂ⇍ື≀ࡢࢽ࣮ࣗࣟࣥ᪂⏕ࢆ

53

ドキュメント内 グリシドールを例としたニューロン発達指標の確立による神経毒性評価に関する研究 (ページ 36-54)