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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81918完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李士傑(Shyh-Jye Lee) | |
| dc.contributor.author | Jin-Jie Liao | en |
| dc.contributor.author | 廖晉頡 | zh_TW |
| dc.date.accessioned | 2022-11-25T03:06:36Z | - |
| dc.date.available | 2026-09-30 | |
| dc.date.copyright | 2021-10-16 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-04 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81918 | - |
| dc.description.abstract | 先天性心臟病(CHD)是世界上最常見的新生兒缺陷，同時伴隨著異常的心臟發育過程以及形態。因此，更深入地了解潛在的致病調節機制對於精確診斷和開發潛在療法至關重要。微核糖核酸(MicroRNAs，miRNA)藉由轉錄後修飾進而調節基因表現以調控器官發育過程中不同的細胞訊息傳導路徑，其中也包括心臟發育。儘管已知某些miRNA會介導心臟發育過程，但許多其他miRNA的心臟相關功能仍然尚未明瞭。先前研究指出，Let-7 miRNA家族其中之一的miR-98在各種類型的心血管疾病皆發現到有表現失調的現象。然而，對於miR-98是否參與心臟發育過程知之甚少。Let-7 miRNA家族已知是早期胚胎發育過程中的關鍵因子，並且與心臟病相關。在斑馬魚中，let-7的過度表達已被證明會延遲胚胎發育並甚至導致死亡。我已經通過注射相似劑量的let-7h模擬物證實了先前報告的發現，而let-7h即是人類miR-98的斑馬魚同源基因。我們進一步發現到注入亞致死劑量的let-7h模擬物並不會導致明顯的早期胚胎缺陷，並且經過let-7h過量表達的胚胎可以在受精後存活長達 72 小時。在這種條件下，我們觀察到let-7h過量表達的斑馬魚胚胎的心室縮小了，但相反的，心房體積卻擴大了。而let-7h的過量表達也降低了各種心臟標誌基因的表現量。此外，通過共同注射幾種預測的let-7h目標基因的mRNA，包括wnt9b、bmp7a和fgf4，可以在某種程度上的挽救這種let-7h所引起的心臟發育缺陷。此外，我們發現let-7h誘導的心臟缺陷是經由lin28ab所調控，同時過量表達lin28ab也對let-7h所引起的心臟缺陷產生顯著的挽救效果。最後，為了闡明let-7h在心臟發生過程中的特定功能，我們建立並驗證了可以在特定時間點以及空間經由tamoxifen誘導的let-7h過量表達基因轉殖魚，以供後續在活體分析let-7h對心臟發育之影響。總結來說，適量的let-7h表現會通過調節許多下游心臟基因表現量維持正常心臟發育過程，同時此等調控機制是經由lin28ab的作用途徑進行調節。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T03:06:36Z (GMT). No. of bitstreams: 1 U0001-2909202115323800.pdf: 28295890 bytes, checksum: dfe645fccec3459e2cfc099c6e95a820 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "口試委員會審定書 i 致謝 ii 中文摘要 ix Abstract xi Introduction 1 Materials and Methods 7 Zebrafish maintenance 7 MicroRNA mimics and mRNA microinjection 7 Imaging 8 Analyses of cardiac functions 8 Total RNA extraction, cDNA preparation and quantitative polymerase chain reaction (qPCR) 9 Plasmid construction 9 Generation of transgenic fish 11 Statistical analysis 11 Results 12 Overexpression of let-7h causes abnormality, developmental delay and death in zebrafish embryos 12 Sub-lethal dosage of let-7h mimic leads to abnormal heart development 13 let-7h mimic disturbs chamber ballooning process and reduces ventricle shortening fraction but not heart rate 14 Down regulation of cardiac genes in the let-7h-overexpressing embryos 16 Partial rescue of let-7h mimic-induced heart defects by the combination of let-7h-target genes 17 let-7h induced heart defect is Lin-28 dependent 19 Generation and screening of transgenic line with tamoxifen-dependent GAL4-UAS expression system 20 Discussion 24 References 33 Table 1. Statistical analysis of the change of embryo morphologies at 24 hpf after sub-lethal dosage let-7h mimic injection. 45 Table 2. Statistical analysis of the change of heart morphologies of let-7h injected embryos at 48 hpf. 46 Table 3. Statistical analysis of the rescue effects of let-7h induced heart defects by co-injecting the mRNAs of different target genes individually. 48 Table 4. Statistical analysis of the rescue effects of let-7h induced heart defects by co-injecting a combination of mRNAs of multiple target genes. 50 Table 5. Statistical analysis of the rescue effects of let-7h-induced heart defects by co-injecting lin28ab mRNA. 53 Table 6. List of all members of Let-7 family in zebrafish 54 Table 7. List of primers 55 Figure 1. Overexpression of let-7h causes developmental defects. 56 Figure 2. Sub-lethal dosages of let-7h mimic interferes cardiac looping and development. 57 Figure 3. Sub-lethal dosages of let-7h mimic disturbs cardiac ballooning process. 58 Figure 4. let-7h mimic does not affect heart rate but reduces ventricle shortening fraction. 59 Figure 5. let-7h mimic induces changes in expression of cardiac-related genes. 61 Figure 6. let-7h mimic-induced heart defects can be partially rescued by co-injecting a combination of mRNAs of multiple target genes. 63 Figure 7. The heart defects induced by let-7h are lin28-dependent. 65 Figure 8. Establishment and validation of spatially and temporally controlled let-7h overexpression transgenic fish. 67 Figure 9. Maps of plasmids used to generate tamoxifen-inducible transgenic lines. 69 Figure 10. Establishments of two transgenic fish lines using the tamoxifen-dependent GAL4-UAS expressing system. 70" | |
| dc.language.iso | en | |
| dc.subject | lin28ab | zh_TW |
| dc.subject | 斑馬魚 | zh_TW |
| dc.subject | 先天性心臟病 | zh_TW |
| dc.subject | let-7 | zh_TW |
| dc.subject | 心臟發育 | zh_TW |
| dc.subject | zebrafish | en |
| dc.subject | Congenital heart disease | en |
| dc.subject | let-7 | en |
| dc.subject | cardiogenesis | en |
| dc.subject | lin28ab | en |
| dc.title | 異位性表達微核糖核酸let-7h干擾斑馬魚心臟發育過程 | zh_TW |
| dc.title | Ectopic expression of let-7h disturbs heart development in zebrafish | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 楊鎧鍵(Hsin-Tsai Liu),詹世鵬(Chih-Yang Tseng),賴時磊,蔡素宜 | |
| dc.subject.keyword | 先天性心臟病,let-7,心臟發育,lin28ab,斑馬魚, | zh_TW |
| dc.subject.keyword | Congenital heart disease,let-7,cardiogenesis,lin28ab,zebrafish, | en |
| dc.relation.page | 70 | |
| dc.identifier.doi | 10.6342/NTU202103457 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2021-10-05 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生命科學系 | zh_TW |
| dc.date.embargo-lift | 2026-09-30 | - |
| 顯示於系所單位： | 生命科學系 | |
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| U0001-2909202115323800.pdf 此日期後於網路公開 2026-09-30 | 27.63 MB | Adobe PDF |
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