RUI: Unraveling the Developmental Genetics that Underlie Anuran Limb Initiation

RUI：揭示阿努拉肢体启动背后的发育遗传学

• 批准号: 2042146
• 负责人: John Young
• 金额: $ 31.58万
• 依托单位: Simmons University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2042146&HistoricalAwards=false
• 关键词: RUI; Unraveling; Developmental; Genetics; Underlie

Most organisms with arms or legs hatch, or are born, with limbs that are largely functional. Accordingly, these animals make their limbs during embryonic development. The study of how limbs develop has taught us much about how cells interact and how changes in developmental processes result in different limb shapes. For example, differences between the leg of a mouse and the wing of a bat can be traced to subtle changes in limb development. Surprisingly missing from our understanding is how animals, such the tadpoles of frogs, are capable of making a limb when they are no longer embryos. This project aims to discover what part of the tadpole body generates the limbs of the frog prior to its metamorphosis. Combining traditional methods with newly available genetic tools this research will reveal what cells in the tadpole generate the limb and how the genes that regulate its development are controlled. The findings from this work will reveal alternative methods by which nature makes the limbs of animals. Further, they will show us the possibilities of growing limbs during different stages of an animal’s life according to the rules of nature. Finally, this work will have impacts beyond scientific discoveries as many of the experiments will be carried out by undergraduates from a woman centered university, student researchers that comprise the next generation of scientists.Studies of the vertebrate limb have provided a deep understanding of the cellular, genetic, and molecular mechanisms that generate an appendage. They have also revealed how the basic pattern of tetrapod limbs can been modified through evolution to give rise to the myriad of arms, legs, wings, and fins we observe in nature. The majority of limb experimentation has been in animals where the limb forms early in embryogenesis. Yet, several tetrapods, most notably frogs, form their limbs well after embryonic patterning and differentiation has occurred. Surprisingly, we know very little about the processes that initially direct limb formation in amphibian tadpoles since most limb research in these animals has focused on regeneration. Recent technological advancements now allow for us to use the frog Xenopus to determine how a tadpole generates a limb when embryonic tissue is no longer present. Overall, this project seeks to answer this question by testing the hypotheses that frogs generate their limbs from multiple embryonic origins and genetically delay the typical tetrapod limb initiation program. This hypothesis will be tested by first determining the embryological origins and induction stage of the limb buds. Then limb gene enhancers will be interrogated to discover how the initiation of frog limbs is controlled. Taken together, this work will provide a fuller understanding of tetrapod limb diversity and plasticity. Finally, the experiments conducted here will be at the forefront of developing new technologies for use by both the Xenopus community as well as those studying the limb more broadly.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

大多数有胳膊或腿的生物在孵化或出生时，四肢基本上是有功能的。因此，这些动物在胚胎发育期间制造四肢。对肢体如何发育的研究教会了我们许多关于细胞如何相互作用以及发育过程中的变化如何导致不同肢体形状的知识。例如，小鼠的腿和蝙蝠的翅膀之间的差异可以追溯到肢体发育的细微变化。在我们的理解中，令人惊讶的是，动物，比如青蛙的蝌蚪，在它们不再是胚胎的时候，是如何能够制造肢体的。这个项目旨在发现蝌蚪身体的哪个部位在青蛙变态之前产生了它的四肢。结合传统方法和新可用的遗传工具，这项研究将揭示蝌蚪中的哪些细胞产生肢体，以及调控其发育的基因是如何控制的。这项工作的发现将揭示大自然制造动物肢体的替代方法。此外，它们将向我们展示根据自然规则在动物生命的不同阶段生长四肢的可能性。最后，这项工作的影响将超越科学发现，因为许多实验将由一所以女性为中心的大学的本科生进行，他们是组成下一代科学家的学生研究人员。对脊椎动物肢体的研究提供了对产生附件的细胞、遗传和分子机制的深刻理解。他们还揭示了如何通过进化来修改四足动物四肢的基本模式，以产生我们在自然界中观察到的无数手臂、腿、翅膀和鳍。大多数肢体实验都是在动物身上进行的，肢体在胚胎发育的早期就形成了。然而，一些四足动物，尤其是青蛙，在胚胎模式和分化发生后很好地形成了它们的四肢。令人惊讶的是，我们对两栖类蝌蚪最初指导肢体形成的过程知之甚少，因为这些动物的大多数肢体研究都集中在再生上。最近的技术进步现在允许我们使用青蛙非洲爪哇来确定当胚胎组织不再存在时，蝌蚪如何产生肢体。总体而言，这个项目试图通过测试以下假设来回答这个问题：青蛙从多个胚胎来源产生四肢，并从基因上推迟典型的四足动物肢体启动程序。这一假设将通过首先确定肢芽的胚胎学起源和诱导阶段来检验。然后，将询问肢体基因增强器，以发现青蛙肢体的启动是如何控制的。综上所述，这项工作将提供对四足动物肢体多样性和可塑性的更全面的了解。最后，这里进行的实验将是开发新技术的前沿，供非洲爪哇社区以及更广泛地研究肢体的人使用。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Facile methods for reusing laboratory plastic in developmental biology experiments

在发育生物学实验中重复使用实验室塑料的简便方法

• DOI: 10.1016/j.diff.2022.11.001
• 发表时间: 2023
• 期刊: Differentiation
• 影响因子: 2.9
• 作者: Clancy, Maggie;Wade, Isabel S.;Young, John J.
• 通讯作者: Young, John J.