Phenotypic flexibility in amphibious fishes

在脊椎动物的进化过程中，由于空气和水的性质截然不同，从生活在水中到迁徙到陆地上的过渡是非常具有挑战性的。化石记录没有保留可能促进这种转变的软组织反应。通过研究今天生活的两栖鱼类，我们可以揭示出我们的鱼类祖先可能出现的各种特征和条件，这些特征和条件使它们能够在水中茁壮成长。总体假设是，血液氧气运输(目标1)、皮肤(目标2)和骨骼肌(目标3)的特征已经进化出有别于非两栖鱼类的特殊特征，以及可逆可塑性在增强鱼类离开水中的功能方面发挥了作用。我们将以自花授粉的两栖红树林鱼类Kyptlebias marmoratus作为模式物种，以及精心挑选的密切相关的两栖和非两栖姐妹对，在适应水和/或空气的鱼类中检验10个新的假设。对于目标1，我们将采集血液样本并分析血红蛋白与氧的结合，对血红蛋白多肽进行测序以确定不同的亚型，并进行整体动物实验以确定呼吸性能。对于目标2，我们将使用组织学方法来确定皮肤细胞类型和蛋白质转运蛋白的定位，以及整个动物实验来确定呼吸和氮排泄的位置。在目标3中，我们将使用组织学和生化方法来确定肌肉纤维类型和调节因素，并进行整体动物研究，以将呼吸和运动性能联系起来。使用这种综合的方法，我们将能够识别使两栖鱼类连接两个截然不同的世界--空气和水--的进化和机械因素。

During the evolution of vertebrates, the transition from living in water to moving onto land was extremely challenging because of the very different properties of air and water. The fossil record has not retained the soft tissue responses that may have facilitated this transition. By studying amphibious fishes living today, we can uncover the variety of traits and conditions that may have arisen in our fish-like ancestors that allowed them to thrive out of water. The overall hypothesis is that blood oxygen transport (goal 1), and skin (goal 2) and skeletal muscle (goal 3) characteristics have evolved specialized features distinct from nonamphibious fishes and as well, reversible plasticity plays a role in enhancing function in fish out of water. We will use the self-fertilizing amphibious mangrove fish Kryptolebias marmoratus as a model species, as well as carefully selected closely-related amphibious and nonamphibious sister pairs to test ten novel hypotheses in fish acclimated to water and/or air. For goal 1, we will collect blood samples and analyze hemoglobin-oxygen binding, sequence hemoglobin peptides to identify different isoforms and conduct whole animal experiments to determine respiratory performance. For goal 2, we will use histological methods to identify skin cell types and protein transporter localization, as well as whole animal experiments to determine sites of respiration and nitrogen excretion. In goal 3, we will use histological and biochemical methods to identify muscle fiber type and regulatory factors, and whole animal studies to link respiratory and locomotory performance. Using this comprehensive approach, we will be able to identify evolutionary and mechanistic factors that have allowed amphibious fishes to bridge two very different worlds, air and water.