利用受精后7天的幼虫,比较自由饲养与限制成像室环境下饲养的日粮摄取量。

Chandler Russell, D. Parnell, J. Orell, J. Walters
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摘要

在美国,心脏病和糖尿病都是十大主要死亡原因,两者都与肥胖有关。由于涉及人类受试者的伦理问题,研究人体代谢可能会很困难,但斑马鱼幼虫(Danio rerio)是透明的。受精后7天的斑马鱼幼虫也已经完全形成了消化道,由于耗尽了蛋黄的供应,它们正在积极寻找新的食物。自由饲养环境通常是向幼虫引入新食物的标准环境,但在其成像方面缺乏通用性。沃尔特斯实验室的微流控芯片可以在幼虫进入食物之前、期间和之后对其进行成像。为了证明这两种饲养环境,我们将荧光消化标记物掺入高脂饲料中进行比较。这些自由喂养的幼虫被放入一个充满饲料的容器中,在28摄氏度的孵化器里放置一个轨道摇床,让它们进食3个小时。三小时后,这些幼虫被冲洗出来并成像。将微流控芯片幼虫装入芯片中饲喂日粮,并在同一台蔡司Discovery V8显微镜下对自由饲喂的幼虫进行成像。对这一数据的未来预测是,微流控芯片内的幼虫比自由饲料中的幼虫消耗的食物略少。这方面的结果仍在进行中,但似乎有助于支持这一假设。
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Comparing dietary intake of free-feeding to feeding in restrained imaging chamber environments using seven days post fertilization larvae.
Heart disease and diabetes are both in the top ten for leading causes of death in the United States with both being linked to obesity. Studying human metabolism can prove difficult due to ethical concerns with human subjects but zebrafish larvae (Danio rerio) are optically transparent. Zebrafish larvae at seven days post fertilization also have fully formed digestive tracts and are actively seeking new food due to exhausting their supply of egg yolk. Free feeding environments are typically the standard when introducing new food to the larvae but lack versatility in their imaging. The Walters lab microfluidic chip can image larvae before, during, and after their introduction to the diet. To prove the two types of feeding environments were compared by using fluorescent digestive markers mixed into high fat diets. The free fed larvae were put into a well filled with diet and allowed to feed for three hours while sat on a orbital shaker inside of a 28 degrees Celsius incubator. After three hours, these larvae were rinsed out and imaged. The microfluidic chip larvae were mounted into the chip and fed the diet while inside and under the same Zeiss Discovery V8 microscope the freely fed larvae were imaged under. Future predictions for this data were that the larvae would consume slightly less diet inside of the microfluidic chip than the larvae in free feed. Results for this are still ongoing but seem to lead towards supporting the hypothesis.
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