- 學位論文
建立ImageJ分析方法來量測班馬魚與水蚤的心跳與血流
Establish a Simple ImageJ-Based Method for Cardiovascular Function Assessment in Fish and Daphnids
摘要
心血管疾病是威脅人類健康的重要疾病之一，它可能導致心臟血液流動中斷，心肌損傷，心律不齊甚至死亡。藥物的非專一性靶向作用可能會引起副作用，其中以心血管毒性是最常發生且受到注意的。因此，在藥物開發過程中，常需要對細胞或是動物模型進行心血管功效與毒性評價。斑馬魚是低階脊椎動物中一個具有許多優勢的新興動物模型，在發育初始階段，斑馬魚胚胎是透明的，利於心臟和血液循環的活體動態觀察。此外，斑馬魚與人類的心臟具有類似的動作電位、分子和生理機制。它還具有與人類相同的封閉式血液循環系統，因此可以利用斑馬魚用作研究人類心血管疾病的簡易模型。水蚤是一種甲殼類無脊椎動物，位於淡水食物鏈中的底層角色。由於生命週期短、容易養殖並且對化學物質敏感，因此是最受歡迎的水生毒性測試動物之一。水蚤也有透明的身體和相對較大的心臟，可以研究毒性物質對其心臟生理的影響，也可以用於預篩選環境毒性試驗。此外，水蚤是第一個完成全基因組測序的甲殼類動物，它們大多數的基因與人類具有同源性。這使得水蚤可用作參考模型，以解決與人類健康直接相關的問題。之前測量心律及血流速率的方法雖然已被建立，但是許多科學家在進行此實驗時，常遇到的限制是需要使用較為昂貴及複雜的設備，有時還須自己寫程式語言來分析數據。此外，藥物治療對其他心血管功能參數，例如心輸出量與血液流速的影響仍有待探索。因此，本論文的主要目的是利用斑馬魚胚胎和水蚤為實驗材料，為心血管功能評估提供低成本和易於設置的軟硬體設備。通過ImageJ開源平台與數學計算公式的建立，我們成功建立了量測斑馬魚和水蚤心血管功能的方法，並利用此方法進行藥物對心血管功能的影響評價
並列摘要
Cardiovascular disease, one of the potential disease which have an impact on human health. It can cause blood flow disruption to the heart, heart muscle injury, heart rate irregularity, and even death. This are rapidly becoming a major threat to the world due to its mortality rate on humans. Drugs and compounds, sometimes have missing-target properties that produce undesired side effects which lead into lethal consequences. Cardiotoxicity is one of the most common issues. Therefore, assessment of the drugs and compounds in animal model is needed. Zebrafish, as a low-level vertebrate is the emerging animal model that has many advantages. In the initial stage, the zebrafish body is transparent and the heart and blood circulation system already fully developed, hence the observation can be done. Moreover, zebrafish shares the same action potential, molecular, and physiological mechanism of heart like human. It also has closed blood circulation system same as human, making it possible to use zebrafish as blood model for human disease. Water fleas, an invertebrate species are among the most favored animals for aquatic toxicity testing due to their short life span, can be easily cultivated, and sensitive to chemicals. It is also a major component of fish zooplankton diet and a key user of freshwater food chains. Water fleas also have transparent body and large hearts, which allows studying the effects of toxicants on their cardiovasular system. Moreover, they are the first crustacean to have their genome sequenced and their most genes are identical to humans. This allows water fleas to be used as a reference model to address issues which directly relevant to human health. Previous methods have been established to calculate the heart rate and blood flow velocity. However, expensive and complicated apparatus is one of the limitations for the other scientists to conduct the experiment. Complicated manual script and programing language is another difficulty. Moreover, the effect of drug treatments on other cardiovascular function parameters, such as stroke volume and cardiac output remains to be explored. Therefore, the specific aim of this thesis is to provide low cost and easy setup for cardiovascular function assessment. By adapting ImageJ as an open source platform and establishing some mathematic formulation, we have successfully established method to measure the cardiovascular function and applied this method to evaluate and quantify the potential adverse effect of chemicals in zebrafish embryos and water fleas.