鈣離子訊號在斑馬魚肌肉發育過程中所扮演之角色

鈣離子的調控對於成體肌肉的影響是被大家所熟知的，例如咖啡因會促使鈣離子釋放進入細胞，促使肌肉細胞發生短期的痙攣,並造成肌肉痠痛及無力。但鈣離子的功能對於肌肉早期發育的影響如今並非如此明瞭。本論文利用2-APB (2-Aminoethoxydiphenyl borate)對於細胞內鈣離子濃度進行調控，進而觀察鈣離子對斑馬於早期肌肉發育所造成的影響。我們的實驗結果指出，在經過浸泡2-APB後之斑馬魚體內鈣離子的數量確實被抑制並使魚體產生彎曲，以及肌肉纖維變異等不正常表型。同時我們經由針對myod所做之原為雜交實驗，發現在浸泡2-APB後，會過度表現與表現位置異常，因此我們利用myod mopholino來降低魚體內其mRNA的表現量,再以2-APB浸泡，可以發現肌肉纖維回復正常的狀態。除此之外，在我們的實驗中，利用咖啡因來提升細胞內鈣離子的濃度，也確實可以挽救2-APB所導致之細胞內鈣離子不足所造成之不正常表型。為了更加確定對於斑馬於早期肌肉發育的調控是由鈣離子濃度所驅動,我們也針對了一些與鈣離子相關並參與肌肉發育和分化的基因做定量-PCR (real-time PCR) ，我們發現desma、fbxo32及nfatc1基因會因2-APB浸泡而表現量上升，反之，cpped1基因會因2-APB浸泡而表現量下降，利用這些證據，我們建立了一套模型，期望藉此得以說明鈣離子訊號在斑馬魚肌肉發育過程中所扮演的角色。

關鍵字

斑馬魚；鈣離子；肌肉

並列摘要

Calcium signaling plays important physiological roles on muscle physiology. For example, enhancement of cellular calcium concentration causes short-term spasm, then make muscle faintness or pain. However, the function of intracellular calcium signaling is unclear during early myogenesis. In this thesis, we used 2-APB(intracellular calcium blocker) and caffeine((intracellular calcium inducer) to regulate intracellular calcium concentration and to investigate the effect caused by calcium during early myogenesis. Our data showed that 2-APB treated zebrafish embryos displayed some malformed phenotypes, such as defective somite and myofribril misalignment. We also found that myod signals were up-regulated and expressed disorderly in the 2-APB-treated embryos. Knock down of myod expression can reverse 2-APB-induced muscle defect phenotypes. Results from real-time PCR showed that desma, fbxo32, and nfatc1 were up regulated but cpped1 was down regulated in 2-APB-treated embryos. On the basis of these observations, we proposed a model to elucidate how intracellular calcium delicate regulates early myogenesis

並列關鍵字

zebrafish ； calcium ； muscle

參考文獻

吳欣儒(民98) LMBRD1參與斑馬魚肌肉發育之分子機轉探討淡江大學碩士論文

黃弈惠(民96) 咖啡因在斑馬魚早期胚胎發育時期引發之畸形淡江大學碩士論文

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國際替代計量

鈣離子訊號在斑馬魚肌肉發育過程中所扮演之角色

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