The gut to gills of ion regulation

我的研究计划的首要目标是了解鱼类和其他水生脊椎动物如何调节它们的盐（离子）和pH值，这对生存至关重要。为了理解在分子水平或离子转运蛋白的机制，我的研究将全动物方法与分子遗传学，生物化学和细胞技术相结合。开发抗体来探测离子转运蛋白的表达是我研究的支柱，我计划通过结合新的基因编辑技术（CRISPR-cas9敲除）来扩展这一研究，以了解更多关于潜在机制的信息。在接下来的五年里，我的研究将探索氢钾atp酶（HKA）的功能作用，它也是负责胃酸化的离子转运蛋白。由HKA引起的胃酸化在脊椎动物（鲨鱼和人类）中是高度保守的，它可以提高蛋白质和磷的消化效率，这对生长和骨骼发育至关重要。尽管胃的重要性，我们提出了胃丧失的悖论，这在脊椎动物的进化史上已经独立发生了多次。我们最近对17种有胃缺失报告的鱼类中的3种进行了研究，结果表明胃的缺失伴随着负责胃消化的基因的缺失。胃丧失的原因有很多，但尚未得到很好的理解，但可能与HKA在胃酸化中的作用有关。此外，HKA有多种变体（类似物），其表达位点位于胃外，参与离子（钾）和pH调节。在鱼类中，鳃是离子和pH调节的一个重要器官。体内钾的调节对神经和肌肉的正常运作也是必不可少的，在这些地方，微小的变化可能会产生巨大的不利影响。拟议的研究计划旨在解决：(1)胃酸化的成本；（二）胃酸化损失的潜在原因和（三）后果；(4)胃基因的丢失与胃的关系；(5) HKA的不同变异在不同动物群体中对离子和pH的调节作用。这些项目将建立在已有的国际合作的基础上，并创造新的合作，从而提高加拿大的研究水平。此外，该计划还为年轻科学家提供高水平的研究技能，这将有助于他们融入加拿大的研究界和工业界。除了提高我们对基因丢失在进化中的作用的理解之外，该计划对加拿大水产养殖业具有明显的实际意义，因为更好地了解胃在消化中的作用可以改进饲料配方，以提高生长和产量，同时减少浪费和污染。

The overarching goal of my research program is to understand how fishes and other aquatic vertebrates regulate their salt (ions) and pH levels, which are vital for survival. In order to understand the mechanisms at play down to the molecular level or the ion transport proteins, my research integrates whole-animal approaches with molecular-genetic, biochemical and cellular techniques. The development of antibodies to probe the expression of ion transport proteins provides the backbone of my research, which I plan to extend by incorporating novel gene editing techniques (CRISPR-cas9 knockouts') to learn more about the underlying mechanisms. In the next five years, my research will explore the functional roles of the hydrogen-potassium ATPase (HKA), which is also the ion transport protein responsible for stomach acidification. Stomach acidification by HKA is highly conserved in vertebrates (sharks to humans) and increases the efficiency of protein and phosphorus digestion that are essential to growth and skeletal development. Despite the importance of the stomach, we are presented with the paradox of stomach loss, which has occurred independently multiple times in vertebrate evolutionary history. Our recent examination of three groups of fishes, out of 17 where stomach loss has been reported, indicates that loss of the stomach is accompanied by loss of the genes responsible for stomach digestion. The causes of stomach loss are many, and not well understood, but are likely related to the role of HKA in stomach acidification. In addition, the HKA has multiple variants (paralogues) with sites of expression outside the stomach that are involved in ion (potassium) and pH regulation. In fishes, the gills are an example of an important organ in ion and pH regulation. Regulation of body potassium is also essential for the proper functioning of nerves and muscles, where small changes can have dramatically adverse effects. The proposed research program aims to address: (1) the costs of stomach acidification; (2) the potential causes and (3) consequences of loss of stomach acidification; (4) the relationship between the loss of the stomach genes and the stomach; and (5) the roles that the different variants of HKA play in ion and pH regulation in different groups of animals. These projects will build upon established international collaborations and create new ones, improving Canada's research profile. In addition, the program has a strong training component for young scientists providing high-level research skills, which will facilitate their integration into Canada's research community and industries. In addition to improving our understanding of the role of gene loss in evolution, the program has obvious practical implications to Canada's aquaculture industry where a better understanding of the stomach's role in digestion can lead to improvements in feed formulations to increase growth and production, while reducing waste and pollution.