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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 閔明源 | zh_TW |
| dc.contributor.advisor | Ming-Yuan Min | en |
| dc.contributor.author | 張筠 | zh_TW |
| dc.contributor.author | Yun Chang | en |
| dc.date.accessioned | 2023-08-16T16:50:37Z | - |
| dc.date.available | 2023-11-09 | - |
| dc.date.copyright | 2023-08-16 | - |
| dc.date.issued | 2023 | - |
| dc.date.submitted | 2023-08-09 | - |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/89031 | - |
| dc.description.abstract | 藍斑核（locus coeruleus，LC）產生相位性放電（phasic firing）對於動物的行為調控至關重要，但對於藍斑核相位性放電的產生機制目前並不是很清楚，過去我們實驗室發現可以通過阻斷薄腦片中的興奮性和抑制性突觸傳遞來抑制和增強類相位活動（phasic-like activity，PLA），並且在藍斑核旁定位到一群抑制性中間神經元，但其僅占和藍斑核連結的中間神經元中的20%，我們推測可能還存在著一群興奮性中間神經元，和抑制性中間神經元一起調控藍斑核相位式放電的產生。
我利用TH-cre和VGluT2-FLP雜交後代的小鼠搭配小麥胚芽凝集素（wheat germ agglutinin，WGA）逆行追蹤，成功標定出和藍斑核有連結的興奮性中間神經元，其佔和藍斑核連結的中間神經元中的10%，並且這群興奮性中間神經元位於藍斑核的內側。另外，我使用VGluT2-cre的小鼠，利用注射可同時在細胞本體表現綠螢光蛋白（green fluorescent protein，GFP）及在突觸表現mRuby的腺病毒，再次證實這群興奮性中間神經元和藍斑核有連結。 更進一步，我利用膜片鉗記錄（patch clamp recording）這群會產生自主放電的興奮性神經元，這群興奮性神經元的電生理訊號會和鈣離子影像訊號同步，而這群被記錄的興奮性神經元放電模式（firing pattern）大多為adaptive，少部分為delay。同時利用生物素（biocytin）的填充，證實這群VGluT2+神經元的軸突會伸入藍斑核中。 由這些實驗結果可以得到，在藍斑核內側有一群和其連結的興奮性中間神經元，這是一群會產生自發性活動的神經元，並且在結構上可以支持調控藍斑核相位性放電的產生。 | zh_TW |
| dc.description.abstract | The locus coeruleus (LC) is crucially involved in the regulation of animal behavior through its generation of phasic firing. However, the mechanism underlying the generation of phasic firing in the locus coeruleus is currently not well understood. In our laboratory's past experiments, we have observed the suppression and enhancement of phasic-like activity (PLA) can be achieved by blocking excitatory and inhibitory synaptic transmission. We also located a population of inhibitory interneurons adjacent to the locus coeruleus, which accounts for only 20% of the interneurons connected to the locus coeruleus.Based on these findings, we speculate that there might be a population of excitatory interneurons, in addition to the inhibitory interneurons, working together to regulate the generation of phasic firing in the locus coeruleus.
By using TH-cre cross VGluT2-FLP transgenic mice and utilizing wheat germ agglutinin (WGA) tracer, we traced back and successfully labeled a population of excitatory interneurons that connected to the locus coeruleus. These excitatory interneurons accounted for 10% of the interneurons connected to the locus coeruleus and were located on the medial side of the locus coeruleus. Furthermore, using VGluT2-cre mice, we injected adeno-associated viruses expressing green fluorescent protein (GFP) in the soma and expressing mRuby in synaptic and filled with biocytin, confirming that there is a connection between this group of excitatory interneurons and the locus coeruleus again. In addition, patch clamp recordings revealed that this population of the excitatory neurons which spontaneously generated burst firing exhibited electrophysiological signals that were synchronized with Calcium imaging. Most of these excitatory neurons displayed an adapted firing pattern, while a small part exhibited delayed. Simultaneously, biocytin filling was used to demonstrate that the axons of this group of excitatory neurons extend into the locus coeruleus. From these experimental results, we can conclude that there is a population of excitatory interneurons located on the medial side of the locus coeruleus. This group consists of neurons that exhibit spontaneous activity and structurally capable of regulating the generation of phasic firing. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2023-08-16T16:50:37Z No. of bitstreams: 0 | en |
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| dc.description.tableofcontents | 口試委員會審定書i
謝辭 ii 中文摘要 iii 英文摘要 iv 目錄 v 第一章 緒論1 第二章 研究目的與實驗設計6 第一節 研究目的與假說6 第二節 研究實驗設計6 第三章 材料和方法10 第一節 動物10 第二節 病毒注射10 第三節 灌流和切片10 第四節 免疫組織化學染色11 第五節 共軛焦顯微鏡影像13 第六節 電生理13 第四章 結果16 第一節 系統及病毒專一性測試16 第二節 藍斑核前中間神經元約有10%屬於興奮性藍斑核中間神經元，並且其位置主要分布於藍斑核內腹側18 第三節 興奮性藍班核前中間神經元之電生理與型態特性21 第五章 結論24 第一節 藍斑核周圍存在著一群和其有突觸連結的興奮性中間神經元24 第二節 和藍斑核連結的興奮性中間神經元可能不只10%25 第三節 與藍斑核連結之興奮性中間神經元的軸突伸入藍斑核中25 第四節 這群興奮性中間神經元的活動模式大多為adaptive27 第五節 這群興奮性中間神經元擁有自主放電的特性27 第六章 總結29 第七章 參考文獻30 第八章 圖36 圖ㄧ、cre-loxp及FLP-FRT之系統測試36 圖二、cre依賴性及FLP依賴性病毒專一性之測試38 圖三、藍斑核細胞和周圍興奮性中間神經元之共定位40 圖四、標定和藍斑核細胞連結之興奮性中間神經元的分布42 圖五、標定和藍斑核細胞連結之興奮性中間神經元的熱區圖45 圖六、內側興奮性中間神經元投射到藍斑核內47 圖七、自主放電之興奮性中間神經元的電訊號及GCaMP訊號同步49 圖八、檢視和藍斑核細胞內側的興奮性中間神經元之電生理特性51 圖九、檢視藍斑核細胞內側的興奮性中間神經元之之神經纖維53 圖十、細胞膜片鉗記錄興奮性中間神經元的位置分布圖55 第九章 表57 表一、實驗所使用小鼠之資訊57 表二、所使用的 AAV之資訊58 表三、在不同品系中各類型細胞表現顆數60 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 鈣離子影像 | zh_TW |
| dc.subject | 小麥胚芽凝集素 | zh_TW |
| dc.subject | 興奮性中間神經元 | zh_TW |
| dc.subject | 相位性放電 | zh_TW |
| dc.subject | 藍斑核 | zh_TW |
| dc.subject | Wheat germ agglutinin | en |
| dc.subject | excitatory interneurons | en |
| dc.subject | Calcium imaging | en |
| dc.subject | Locus coeruleus | en |
| dc.subject | phasic firing | en |
| dc.title | 定位小鼠中參與藍斑核活性的調節之興奮性中間神經元群體 | zh_TW |
| dc.title | Identification of a Population of Excitatory Interneurons Involved in the Regulation of Locus Coeruleus Activation in Mice | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 111-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.coadvisor | 陳瑞芬 | zh_TW |
| dc.contributor.coadvisor | Ruei-Feng Chen | en |
| dc.contributor.oralexamcommittee | 陳示國;楊琇雯;傅毓秀 | zh_TW |
| dc.contributor.oralexamcommittee | Shih-Kuo Chen;Hsiu-Wen Yang;Yu-Show Fu | en |
| dc.subject.keyword | 藍斑核,相位性放電,興奮性中間神經元,小麥胚芽凝集素,鈣離子影像, | zh_TW |
| dc.subject.keyword | Locus coeruleus,phasic firing,excitatory interneurons,Wheat germ agglutinin,Calcium imaging, | en |
| dc.relation.page | 60 | - |
| dc.identifier.doi | 10.6342/NTU202303252 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2023-08-10 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 生命科學系 | - |
| 顯示於系所單位： | 生命科學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-111-2.pdf 未授權公開取用 | 15.77 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。