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，因为它具有丰富的发育和遗传信息以及可用于该模型生物的遗传学工具（例如突变体，基因敲除方法，微阵列）。 此外，斑马鱼的世代时间很短，并且该物种在人工饲养中很容易繁殖，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，因为这种物种的体积小（成年体重<500毫克），无法使用大多数标准的生理方法（例如血液采样）。 为了应对这一挑战，我们建议购买一个pH统计滴定设置，这将使我们能够测量净酸排泄个别成年斑马鱼或组斑马鱼胚胎在真实的时间。 我们还建议购买一台仪器（PWee 50），用于测量微量血液样本（需要1微升血液）的血红蛋白-氧结合亲和力。 再加上我们已经开发的测量斑马鱼通气和代谢率的方法，这些新仪器将使我们能够评估这种模式物种的酸碱平衡，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.