Assessing defects in embryonic development due to loss of protein fucosylation

层级摘要 蛋白质岩藻糖基化是一个进化保守的过程，在所有动物中都存在。蛋白质岩藻糖化是指将糖(岩藻糖)分子加到靶蛋白质上，以确保其正常的功能。我们使用小咸水鱼(Danio Rerio)来研究当蛋白质岩藻糖基化被破坏时，胚胎会发生什么。当这些鱼的蛋白质岩藻糖基化所需的基因发生突变时，它们的胚胎出现了脑出血。这表明，蛋白质岩藻糖基化是建立稳定的、功能正常的血管所必需的。我们用斑马鱼来试图弄清楚基因和蛋白质岩藻糖基化是如何构建血管所必需的。 除了脑出血，蛋白质岩藻糖基化缺陷的斑马鱼还有许多其他问题。例如，它们的尾巴是弯曲的，它们游来游去。这些问题和脑出血都可以用纤毛缺陷来解释。纤毛是人体几乎所有细胞上的重要结构，对胚胎的正常发育至关重要。例如，纤毛可以击打和移动液体通过脊柱，而这些纤毛的缺陷被认为是导致鱼类弯曲尾巴的原因。斑马鱼身体外部也有纤毛，可以探测水流的变化，这些纤毛的缺陷可能会导致循环游泳行为。最后，纤毛指向我们的血管，并在血管发育和血压调节中发挥重要作用。众所周知，这些纤毛的缺陷会导致脑出血。在我的实验室里，我们能够看到缺乏蛋白质岩藻糖基化的突变斑马鱼的纤毛，看看是否有任何缺陷。我们观察血管、脊柱和身体外部的纤毛，以确定纤毛缺陷是否导致蛋白质岩藻糖基化缺陷斑马鱼的脑出血、弯曲尾巴和圆形游泳行为。 这项工作对帮助我们了解纤毛是如何发挥作用以及它们为什么重要，并帮助我们了解它们在人类中可能是如何重要的是重要的。例如，导致斑马鱼尾巴弯曲的纤毛缺陷可能会导致人类脊柱侧弯。人类的耳朵里有纤毛帮助我们听力，在结构上几乎与斑马鱼外部帮助它们游泳的纤毛相同。由于纤毛问题，人类患脑血管缺陷的风险也会增加，因此我们的斑马鱼是了解导致中风等血管疾病的潜在生物学问题的有用工具。最后，由于所有动物都需要蛋白质岩藻糖化，我们的工作可以跨越科学学科，为我们提供线索，了解其他动物如何利用蛋白质岩藻糖化来维持正常的身体功能。

Lay Summary Protein fucosylation is an evolutionary conserved process that occurs in all animals. Protein fucosylation refers to the addition of sugar (fucose) molecules to target proteins in order to ensure their proper function. We use the small brackish fish, (Danio rerio) to study what happens to an embryo when protein fucosylation is disrupted. Upon making a mutation in a gene required for protein fucosylation in these fish, their embryos developed cerebral hemorrhages. This indicates that protein fucosylation is required to build stable, functioning blood vessels. We use the zebrafish to try and figure how the gene, and protein fucosylation in general, is required to build blood vessels. Zebrafish with defects in protein fucosylation have a number of other problems in addition to cerebral hemorrhages. For example, their tails are bent and they swim in circles. These problems, and the cerebral hemorrhages, can all be explained by defective cilia. Cilia are important structures located on almost all of our bodies cells, and are critically important for normal embryo development. Cilia for example, can beat and move fluid through the spinal columns, and defects in these cilia are known to cause bent tails in fish. Zebrafish also have cilia on the outside of their body that detect changes in water currents, and defects in these cilia can cause circular swimming behavior. Finally, cilia point into our blood vessels, and play an important role in vessel development and the regulation of blood pressure. Defects in these cilia are known to cause cerebral hemorrhages. In my lab, we are able to visualize cilia in our mutant zebrafish that lack protein fucosylation, to see if there are any defects. We look at the cilia in blood vessels, in the spinal column, and on the outside of the body to see if defective cilia are responsible for the cerebral hemorrhages, bent tails, and circular swimming behavior in our protein fucosylation deficient zebrafish. This work is important to help us understand how cilia function and why they are important, and help us to learn about how they might be important in humans. For example, defects in cilia that cause bent tails in zebrafish can cause scoliosis (bent spines) in humans. Humans have cilia in our ears that help us hear, and are almost identical in structure to the cilia on the outside of zebrafish that help them swim. Humans can also be at increased risk for cerebral blood vessel defects due to problems with cilia, so our zebrafish serve as a useful tool to understand the underlying biological problems that lead to vascular diseases such as stroke. Lastly, as all animals require protein fucosylation, our work can translate across scientific disciplines to give us clues as to how other animals use protein fucosylation in order to maintain normal body function.