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標題: | 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 Identification of a Population of Excitatory Interneurons Involved in the Regulation of Locus Coeruleus Activation in Mice |
作者: | 張筠 Yun Chang |
指導教授: | 閔明源 Ming-Yuan Min |
關鍵字: | 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, Locus coeruleus,phasic firing,excitatory interneurons,Wheat germ agglutinin,Calcium imaging, |
出版年 : | 2023 |
學位: | 碩士 |
摘要: | 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。 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. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/89031 |
DOI: | 10.6342/NTU202303252 |
全文授權: | 未授權 |
顯示於系所單位: | 生命科學系 |
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