斑馬魚是一個研究基因發育及藥物測試的模式生物，基因與人類的相似度有87%，其快速的發育得以壓低成本和進行大量實驗，而且胚胎為體外發育而且透明易觀察，因此現今研究許多研究基因功能的科學家以斑馬魚作為實驗動物。細胞凋亡是一種計畫性死亡，能夠清除不必要或不正常且無法恢復的細胞，細胞凋亡後會產生凋亡小體由磷脂醯絲胺酸受體(PSR)清除。磷脂醯絲胺酸受體是一種位於巨噬細胞上用於辨認凋亡細胞上的磷脂醯絲胺酸來進行吞噬的受體。先前研究發現磷脂醯絲胺酸受體缺陷的斑馬魚之胚胎早期發育出現凋亡細胞堆積在體節的情形，並且發現到磷脂醯絲胺酸受體剃除斑馬魚發受精後 8 小時胚胎發育遲緩而且氧活性分子濃度提升，並在 24 小時腦部、眼睛、心臟及體節發育不正常的情形。本實驗以 CRISPR/Cas9 系統建立磷脂醯絲胺酸受體剃除的 F0 及F1 斑馬魚來研究細胞自噬作用及中腦後腦發育影響。接著利用次世代定序來觀察基因的變化，分析數據後挑出氧活性分子相關基因、細胞自噬基因以及中後腦發育基因，以 qRT-PCR 做表現量確認與次世代定序結果一致。西方點墨法中細胞自噬活動明顯降低，像 ATG5-12 及 Beclin-1 表現相對對照組降低。透過顯微注射補足 PSR mRNA， ROS 的應激反應和自噬相關基因下調被阻斷，腦部發育相對正常。綜合結果發現磷脂醯絲胺酸受體能夠藉由調控凋亡小體來克服氧化
壓力，進而能讓細胞自噬系統正常運作使中腦以及後腦能正常發育。

Zebrafish model system is important in genetic and developmental study, with fully established online gene library available for gene function study, short development period and transparent embryo make researchers fully access to all development stage with very low cost, this make zebrafish a power model in gene study. Phosphatidylserine receptor (PSR) locate on macrophage can identify apoptotic cell s which present phosphatidylserine that originally locate on inner membrane on cell surface. Previous studies show that mouse without PSR have defect in brain, eye and heart development, PSR knock down in zebrafish embryo have identical result. In our previous study, PSR knock out zebrafish line was established by CRISPR/Cas9 system targeting in PSR gene on exon 3.Increased reactive oxygen species was detected in PSR knockout zebrafish embryo during gastrulation stage, swelling heart and smaller brain at 24 hour post fertilization (hpf). In this study, we investigate the gene profile in PSR knockout zebrafish embryo by CRISPR/Cas9 approach. Then we use the NGS analysis at 8 hpf for further evaluation. We found that mid-brain and hindbrain defected embryos have induced ROS-mediated stress signal and stress-related genes . Then, the novel genes downregulation of autophagy-related genes such as Beclin-1, ATG5 and ATG8. Finally, the brain-defect embryos was rescued by extra mRNA of PSR by microinjection that ROS-mediated stress response and downregulation of autophagy-related genes is blocked. Our results provide new insights in PSR-mediated engulfing signal for brain development.

白植友，斑馬魚磷脂醯絲胺酸受器啟動子之選殖與功能分析，國立成功大學生物科技研究所碩士論文，2007。

唐婉倫，利用 CRISPR/Cas9 系統建立斑馬魚磷脂醯絲胺酸受體及 Bad 基因剔除魚隻及其胚胎發育功能分析，國立成功大學生物科技研究所碩士論文，2017。

黃培軒，探討斑馬魚之磷脂醯絲胺酸受器其吞噬死亡細胞能力與否對早期胚胎發育之影響，國立成功大學生物科技研究所碩士論文，2015。

黃冠榮，斑馬魚磷脂醯絲胺酸受器於骨骼發育過程中的影響，國立成功大學生物科技研究所碩士論文，2009。

薛采晴，傳染性脾臟及腎臟壞死病毒(ISKNV)誘導魚類細胞株自噬作用之研究，國立成功大學生物科技研究所碩士論文，2017。

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