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題名 促進成年海馬迴神經前驅細胞增殖的藥物篩選
Promoting proliferation of adult hippocampal neural
作者 魏志安
貢獻者 賴桂珍
魏志安
關鍵詞 海馬迴
成年神經新生
神經球
神經前驅細胞
hippocampus
Neural progenitor cells
neurospheres
Adult neurogenesis
日期 2013
上傳時間 13-Jul-2015 11:14:31 (UTC+8)
摘要 在成年的哺乳類動物大腦中有兩個區域，可以不斷的有新的神經細胞生成，一個位於大腦側腦室旁內側(Subventricular zone of anterior lateral ventricle ;SVZ)，另一個位於海馬迴(hippocampus)內的齒狀迴(Subgran-
ular zone of dentate gyrus ;SGZ) ，其中海馬迴是本論文主要探討的腦區。
神經前驅細胞(Neural progenitor cells :NPC)因具有自我更新(self
-renewal）、增殖(proliferative）、多能(multipotent）的能力以及遷移性(Migration)，所以可利用海馬迴內生性的神經前驅細胞(NPC)，促進其增殖以替代因損傷、老化或疾病而損失的神經細胞。神經前驅細胞經由細胞體外培養過程會形成神經球(Neurospheres)，神經球和神經前驅細胞同樣具有自我更新以及可以分化成其他神經細胞的能力。
本研究觀察到，對成年神經新生進行體外藥物的篩選中，化合物Chemical-X，有明顯的促進神經新生的能力。實驗中取健康成年雄性大鼠為實驗動物，分離出成年大鼠之海馬迴神經前驅細胞。用Chemical-X處理後，觀察神經球自我更新能力，以及再把新生成的神經球利用免疫螢光染色處理，瞭解神經前驅細胞經藥物處理後所新生成的細胞，是否仍維持在神經前驅細胞的狀態。進而評估藥物能否達到促進神經新生的目的。
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Aizawa K, Ageyama N, Terao K, Hisatsune T (2011) Primate-specific alterations in neural stem/progenitor cells in the aged hippocampus. Neurobiology of aging 32:140-150.
Albeck DS, Sano K, Prewitt GE, Dalton L (2006) Mild forced treadmill exercise enhances spatial learning in the aged rat. Behavioural Brain Research 168:345-348.
Altman J (1962) Are new neurons formed in the brains of adult mammals? Science 135:1127-1128.
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Altman J, Das GD (1965b) Post-natal origin of microneurones in the rat brain. Nature 207:953-956.
Alvarez-Buylla A, Garcia-Verdugo JM (2002) Neurogenesis in adult subventricular zone. The Journal of neuroscience : the official journal of the Society for Neuroscience 22:629-634.
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描述 碩士
國立政治大學
神經科學研究所
100754002
102
資料來源 http://thesis.lib.nccu.edu.tw/record/#G0100754002
資料類型 thesis
dc.contributor.advisor 賴桂珍zh_TW
dc.contributor.author (Authors) 魏志安zh_TW
dc.creator (作者) 魏志安zh_TW
dc.date (日期) 2013en_US
dc.date.accessioned 13-Jul-2015 11:14:31 (UTC+8)-
dc.date.available 13-Jul-2015 11:14:31 (UTC+8)-
dc.date.issued (上傳時間) 13-Jul-2015 11:14:31 (UTC+8)-
dc.identifier (Other Identifiers) G0100754002en_US
dc.identifier.uri (URI) http://nccur.lib.nccu.edu.tw/handle/140.119/76462-
dc.description (描述) 碩士zh_TW
dc.description (描述) 國立政治大學zh_TW
dc.description (描述) 神經科學研究所zh_TW
dc.description (描述) 100754002zh_TW
dc.description (描述) 102zh_TW
dc.description.abstract (摘要) 在成年的哺乳類動物大腦中有兩個區域，可以不斷的有新的神經細胞生成，一個位於大腦側腦室旁內側(Subventricular zone of anterior lateral ventricle ;SVZ)，另一個位於海馬迴(hippocampus)內的齒狀迴(Subgran-
ular zone of dentate gyrus ;SGZ) ，其中海馬迴是本論文主要探討的腦區。
神經前驅細胞(Neural progenitor cells :NPC)因具有自我更新(self
-renewal）、增殖(proliferative）、多能(multipotent）的能力以及遷移性(Migration)，所以可利用海馬迴內生性的神經前驅細胞(NPC)，促進其增殖以替代因損傷、老化或疾病而損失的神經細胞。神經前驅細胞經由細胞體外培養過程會形成神經球(Neurospheres)，神經球和神經前驅細胞同樣具有自我更新以及可以分化成其他神經細胞的能力。
本研究觀察到，對成年神經新生進行體外藥物的篩選中，化合物Chemical-X，有明顯的促進神經新生的能力。實驗中取健康成年雄性大鼠為實驗動物，分離出成年大鼠之海馬迴神經前驅細胞。用Chemical-X處理後，觀察神經球自我更新能力，以及再把新生成的神經球利用免疫螢光染色處理，瞭解神經前驅細胞經藥物處理後所新生成的細胞，是否仍維持在神經前驅細胞的狀態。進而評估藥物能否達到促進神經新生的目的。		zh_TW
dc.description.tableofcontents 中文摘要………………………………………………………………Ⅰ
英文摘要………………………………………………………………..Ⅱ
目錄……………………………………………………………………Ⅳ
圖次……………………………………………………………………Ⅷ
表次……………………………………………………………………ⅩV
縮寫對照表……………………………………………………………….…ⅩⅣ
第一章、緒論………………………………………………………… 1
第一節 成年神經新生(Adult neurogenesis)…………………… 1
一、成年神經新生（Adult neurogenesis）的區域…..………… 1
第二節 海馬迴（Hippocampus）構造………………………… 3
一、海馬迴的學習記憶功能………………………………………. 4
二、神經前軀細胞(Neural progenitor cells, NPC)特性………... 5
三、成年神經新生發育過程的細胞標記(Cell maker)……….….. 6
第三節 影響成年神經新生的因素………………………………… 7
一、腦內微環境 (niche)…………………………………………. 7
二、環境（Environmental）.……………………………………..8
三、荷爾蒙（hormones）….………………….…………………9
四、運動（Exercise）.…..…….……………………………....11
五、學習 (Learning)….………………………………………….11
六、老化 (Aging)……………..………………………………….12
七、藥物 (Drug)…………..……………………………………13
第四節 天然化合物…………………………..…………………………..15
一、厚朴的介紹….…………………………...………………....15
二、和厚朴酚藥理作用…….……..….…………….…….…......16
第五節 研究目的與策略…………….……………………………………17
第二章、實驗材料與研究方法..……………………………………………...18
第一節 實驗材料…………………………………………………………...18
一、細胞培養材料………….…………………..…………..……18
二、化學藥品……………………………………….……………18
三、抗體..………………..………..…..……………….………...20
四、實驗儀器.…………………………………………………….20
五、 軟體…………………………………..…………….……….21
六、實驗藥品配製…………………...…………………………..21
七、實驗動物..……………………………………..…………….22
第二節 成年海馬迴神經前驅細胞(neural progenitor cells)分離
培養………………………………………………….............… 23
一、海馬迴神經前驅細胞的分離………………………………...23
二、細胞計數…………...…………………………………………24
三、神經球(neurosphere)的計算……………………………….24
四、細胞培養皿塗層……………………………………….……24
五、培養基的配置……….………….. ……………….………... 25
六、酵素的製備(Enzyme cocktail)…..…….…………………...25
七、Percoll的製備…..……………………………….…….……26
八、phosphate buffer saline(PBS)的製備……….……………26
九、DPBS-glucose平衡鹽溶液的製備……………………..…26
十、Harvest medium(HM)的製備……………………………...26
第三節 免疫螢光染色(Immunofluorescence)................….…….…….…27
一、細胞免疫螢光染色 .…..…….……..………………………27
二、IdU, CldU 細胞免疫螢光染色…..….………………..……28
三、封片膠的製備………..……….…..……..…………….……28
第四節 細胞螢光強度分………………………………….….……………..28
一、Nestin/GFAP螢光強度分析………….……………………29
二、IdU/CldU螢光強度分析…….…...………………....……29
第五節 實驗數據分析…………………………………...…………………30
第三章 結果…………………………………………...………………………31
第一節 分離培養的神經前驅細胞(NPC)組成及分期……..……….....…31
第二節 Microglia的分離與神經前驅細胞(NPC)純度的提高………..…36
第三節 和厚朴酚對體外神經前驅細胞(NPC)的影響……………………39
第四節 Sonic hedgehog對體外神經前驅細胞(NPC)的影響………......52
第五節 Chemical-X對體外神經前驅細胞(NPC)的增殖分析…………..59
第六節Chemical-X藥物對體外神經前驅細胞(NPC)的增殖後分析.…..73
第四章 討論……………………………………………………………..……77
第五章 結論……..……………………………………………………………80
參考資料 …………………………………………………………………....81		zh_TW
dc.source.uri (資料來源) http://thesis.lib.nccu.edu.tw/record/#G0100754002en_US
dc.subject (關鍵詞) 海馬迴zh_TW
dc.subject (關鍵詞) 成年神經新生zh_TW
dc.subject (關鍵詞) 神經球zh_TW
dc.subject (關鍵詞) 神經前驅細胞zh_TW
dc.subject (關鍵詞) hippocampusen_US
dc.subject (關鍵詞) Neural progenitor cellsen_US
dc.subject (關鍵詞) neurospheresen_US
dc.subject (關鍵詞) Adult neurogenesisen_US
dc.title (題名) 促進成年海馬迴神經前驅細胞增殖的藥物篩選zh_TW
dc.title (題名) Promoting proliferation of adult hippocampal neuralen_US
dc.type (資料類型) thesisen
dc.relation.reference (參考文獻) Åberg MAI, Åberg ND, Hedbäcker H, Oscarsson J, Eriksson PS (2000) Peripheral Infusion of IGF-I Selectively Induces Neurogenesis in the Adult Rat Hippocampus. The Journal of Neuroscience 20:2896-2903.
Aizawa K, Ageyama N, Terao K, Hisatsune T (2011) Primate-specific alterations in neural stem/progenitor cells in the aged hippocampus. Neurobiology of aging 32:140-150.
Albeck DS, Sano K, Prewitt GE, Dalton L (2006) Mild forced treadmill exercise enhances spatial learning in the aged rat. Behavioural Brain Research 168:345-348.
Altman J (1962) Are new neurons formed in the brains of adult mammals? Science 135:1127-1128.
Altman J (1963) Autoradiographic investigation of cell proliferation in the brains of rats and cats. The Anatomical record 145:573-591.
Altman J, Das GD (1965a) Autoradiographic and histological evidence of postnatal hippocampal neurogenesis in rats. The Journal of comparative neurology 124:319-335.
Altman J, Das GD (1965b) Post-natal origin of microneurones in the rat brain. Nature 207:953-956.
Alvarez-Buylla A, Garcia-Verdugo JM (2002) Neurogenesis in adult subventricular zone. The Journal of neuroscience : the official journal of the Society for Neuroscience 22:629-634.
Alvarez-Buylla A, Lim DA (2004) For the long run: maintaining germinal niches in the adult brain. Neuron 41:683-686.
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