Repeated Blood Collection for Blood Tests in Adult Zebrafish.
Repeated Blood Collection for Blood Tests in Adult Zebrafish.
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DOI：
10.3791/53272
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发表时间：
2015-08
期刊：
影响因子：
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通讯作者：
Liqing Zang;Yasuhito Shimada;Y. Nishimura;Toshio Tanaka;Norihiro Nishimura
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文献类型：
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作者：
Liqing Zang;Yasuhito Shimada;Y. Nishimura;Toshio Tanaka;Norihiro Nishimura
重复采血是对实验动物进行的最常见的技术之一。然而，从斑马鱼身上采集血液的非致死方案尚未建立。以往的斑马鱼采血方法，如侧边切开、断头和尾部消融等，都是致命的。因此，我们开发了一种新的“重复”采血方法，并在此提出了一份详细的规程。这种方法是微创的，对斑马鱼的死亡率非常低（2.3%），因此可以从同一个体重复采集血液。最大采血量取决于鱼的体重。每周一次重复采血的体积应≤体重的0.4%或每2周≤1%，通过测定血红蛋白来评估。此外，还测量了雄性和雌性成年斑马鱼的血红蛋白、空腹血糖、血浆甘油三酯（TG）和总胆固醇水平。我们还应用该方法研究了饮食引起的肥胖中葡萄糖代谢的失调。这种血液采集方法将允许许多应用，包括葡萄糖和脂质代谢和血液学研究，这将增加斑马鱼作为人类疾病模式生物的使用。
Repeated blood collection is one of the most common techniques performed on laboratory animals. However, a non-lethal protocol for blood collection from zebrafish has not been established. The previous methods for blood collection from zebrafish are lethal, such as lateral incision, decapitation and tail ablation. Thus we have developed a novel "repeated" blood collection method, and present here a detailed protocol outlining this procedure. This method is minimally invasive and results in a very low mortality rate (2.3%) for zebrafish, thus enabling repeated blood sampling from the same individual. The maximum volume of blood sampling is dependent on body weight of the fish. The volume for repeated blood sampling at intervals should be ≤0.4% of body weight every week or ≤1% every 2 weeks, which were evaluated by measurements of blood hemoglobin. Additionally, hemoglobin, fasting blood glucose, plasma triacylglycerol (TG) and total cholesterol levels in male and female adult zebrafish were measured. We also applied this method to investigate the dysregulation of glucose metabolism in diet-induced obesity. This blood collection method will allow many applications, including glucose and lipid metabolism and hematological studies, which will increase the use of zebrafish as a human disease model organism.