Micro-analysis system for Zebrafish studies

斑马鱼为生理功能的研究提供了一个强大的模型，因为斑马鱼有丰富的发育和遗传信息以及遗传学工具（如突变体、基因敲低方法、微阵列）。此外，斑马鱼的繁殖时间很短，在圈养环境下很容易繁殖，因此可以使用实验进化或人工选择等方法。然而，斑马鱼的生理数据很少，因为该物种体积小（成年质量<500毫克），无法使用大多数标准的生理方法（如血液采样）。为了解决这一挑战，我们建议购买一个pH值滴定装置，使我们能够实时测量单个成年斑马鱼或斑马鱼胚胎组的净酸排泄。我们还建议购买一种仪器（PWee50），可以在微量血液样本上测量血红蛋白-氧结合亲和力（需要1 μ L血液）。再加上我们已经开发的测量斑马鱼通风和代谢率的方法，这些新仪器将使我们能够评估这种模型物种的酸碱平衡、离子调节和气体转移。测量生理功能的能力将使我们能够利用斑马鱼模型的力量，一方面将生理功能与基因表达联系起来，另一方面将其与适应（进化）联系起来。

Zebrafish provide a powerful model for studies of physiological function because of the wealth of developmental and genetic information as well as genetics tools (e.g. mutants, gene knockdown approaches, microarrays) that are available for this model organism. In addition, the generation time in zebrafish is short and the species breeds readily in captivity, allowing approaches such as experimental evolution or artificial selection to be used. However, physiological data for zebrafish are sparse because the small size of this species (adult mass <500 mg) precludes the use of most standard physiological approaches (e.g. blood sampling). To address this challenge, we propose to purchase a pH stat titration set-up that will allow us to measure net acid excretion by individual adult zebrafish or by groups of zebrafish embryos in real time. We also propose to purchase an instrument (PWee50) that allows haemoglobin-oxygen binding affinity to be measured on minute blood samples (1 µL of blood is needed). Coupled with the methods that we have already developed to measure ventilation and metabolic rate in zebrafish, these new instruments will enable us to assess acid-base balance, ionic regulation and gas transfer in this model species. The ability to measure physiological function will allow us to harness the power of the zebrafish model for linking physiological function to gene expression on the one hand, and to adaptation (evolution) on the other.