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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 潘建源(Chien-Yuan Pan) | |
| dc.contributor.author | Hua-Hua Wu | en |
| dc.contributor.author | 吳畫畫 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:08:33Z | - |
| dc.date.available | 2021-11-03 | |
| dc.date.available | 2022-11-24T03:08:33Z | - |
| dc.date.copyright | 2021-11-03 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-26 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80525 | - |
| dc.description.abstract | "鈣訊號參與於多種細胞活性如神經傳遞，突觸可塑性，以及學習記憶等高階腦功能。有一群與calmodulin結構類似的鈣離子結合蛋白，但在4個EF-hand motifs之中，僅有2-3個具有鈣離子結合能力。Calneuron 1（Caln1）僅在N端有2個具功能的EF-hand motifs，和一個在C端幫助定位於膜上的跨膜疏水段。我們先前實驗已證實Caln1主要表現於成鼠腦中，而非胚胎，且可削弱鈣離子通道活性，但Caln1如何影響神經分化仍未知。為了探討Caln1在神經發育上的功能，我們將Caln1和其突變體，表現於初級培養的大鼠胚胎皮質神經細胞中，結果顯示，喪失疏水段的突變體，能夠增加神經纖維的數量，並增強谷氨酸誘導的鈣離子反應。為了進一步探究Caln1在神經分化中扮演的角色，我們將其轉染於有或無 NGF處理的PC12細胞；結果顯示，Caln1的過表達，會拮抗NGF增加細胞直徑與鈣離子上升反應的效果，因此Caln1在調控神經分化和鈣離子濃度恆定中，可扮演關鍵角色。為了解Caln1在腦發育中的功能，我們以RT-PCR分析斑馬魚胚胎不同發育時期， caln1同源基因 (z-caln1和z-caln2) 的表現。該結果顯示，選擇性剪接的異構型z-caln1-X2和z-caln2-X2，在胚胎發育早期便有高表現量，而其他異構型則是24 hpf後才有顯著表現。因此我們將z-caln1-X2和z-caln2-X2這兩個異構型亞克隆進載體，並設計嗎啉代寡核苷酸（ morpholino oligos, MOs）以探討降低或增加表現，对斑馬魚胚胎發育的影響。已有報導指出，Caln1與思覺失調症高度相關，我們的結果不僅說明Caln1如何調控神經發育，並發展相關病變的可能治療方式。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:08:33Z (GMT). No. of bitstreams: 1 U0001-2610202116593800.pdf: 2191951 bytes, checksum: d3bb88aed233c9c7a58c04b0ad958ecd (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | Acknowledgement i 摘要 ii Abstract iii 1. Introduction 1 1.1. Neuronal calcium signaling 1 1.2. CaM superfamily and EF-hand calcium binding motif 2 1.3. The basic functions of CaM and CaBP family 4 1.4. Caln1 structure in human and rat 6 1.5. The basic functions and distributions of Caln1 in human and rat 7 1.6. Caln1 expression patterns in zebrafish early development 9 1.7. Regulation of neuronal development and neuronal differentiation by calcium signaling 10 1.8. NGF in initiating neuronal differentiation 12 1.9. Cultured rat embryonic neurons and zebrafish embryos as neuronal differentiation models 12 2. Aims 14 3. Materials and Methods 15 3.1. Chemicals and solutions/buffer 15 3.2. Primary cultured neurons and PC12 cells preparation 15 3.3. Zebrafish embryos collection 16 3.4. Plasmid construction 16 3.5. Transfection 17 3.6. Fluorescence imaging 17 3.7. Ca2+ imaging 18 3.8. RNA extraction and RT-PCR (primers design) 20 3.9. Data analysis 21 4. Results 22 4.1. Alignment of Caln1 in different species 22 4.2. Caln1ΔHT enhances neurite outgrowth in cultured cortical neurons 23 4.3. Caln1ΔHT enhances glutamate-induced Ca2+ response in cortical neurons 24 4.4. Caln1 overexpression decreases PC12 cell size 26 4.5. Caln1 enhances Ca2+ response in ATP-stimulated PC12 cells 26 4.6. Z-caln1 and z-caln2 isoforms are differentially expressed during the development zebrafish embryos 27 4.7. MO sequences are designed for z-caln1-X2 and z-caln2-X2 knockdown 29 4.8. Construct expression vector to overexpress z-caln1-X2 and z-caln2-X2 in zebrafish embryos 29 5. Discussion 30 5.1. Caln1 and neuron differentiation 30 5.2. Effects of Caln1 on Ca2+ responses 31 5.3. Caln1 might play an important role of brain development in zebrafish 32 6. Conclusion 33 7. References 34 Table. 43 Figures 45 | |
| dc.language.iso | en | |
| dc.subject | 嗎啉代寡核苷酸 | zh_TW |
| dc.subject | 鈣離子結合蛋白 | zh_TW |
| dc.subject | PC12細胞 | zh_TW |
| dc.subject | 神經發育 | zh_TW |
| dc.subject | 鈣離子穩態 | zh_TW |
| dc.subject | PC12 cell | en |
| dc.subject | Ca2+-binding protein | en |
| dc.subject | Ca2+ homeostasis | en |
| dc.subject | Calneuron 1 | en |
| dc.subject | morpholino | en |
| dc.subject | neuron development | en |
| dc.title | 探討 Calneuron I 在大鼠胚胎皮質神經細胞及斑馬魚腦發育中的功能 | zh_TW |
| dc.title | Study the Functions of Calneuron I in the Development of Cultured Rat Embryonic Cortical Neurons and Zebrafish Brain | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 周銘翊(Hsin-Tsai Liu),周申如(Chih-Yang Tseng) | |
| dc.subject.keyword | 鈣離子結合蛋白,鈣離子穩態,嗎啉代寡核苷酸,神經發育,PC12細胞, | zh_TW |
| dc.subject.keyword | Ca2+-binding protein,Ca2+ homeostasis,Calneuron 1,morpholino,neuron development,PC12 cell, | en |
| dc.relation.page | 61 | |
| dc.identifier.doi | 10.6342/NTU202104252 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-10-27 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生命科學系 | zh_TW |
| 顯示於系所單位： | 生命科學系 | |
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