Identification of molecular mechanisms regulating skeletal morphology

脊椎动物的骨骼提供了多种重要的功能，包括形状、强度、柔韧性和保护重要器官。复杂形式的骨骼的建立需要生长与骨形成细胞和关节形成细胞的分化的协调。 事实上，关于细胞如何分裂以及相关细胞类型如何分化，我们已经知道了很多。然而，很少有人知道这些事件是如何协调的，以产生正确大小和形状的骨骼元素。差距连接蛋白Cx43已被证明在斑马鱼鳍再生过程中调节细胞增殖和关节形成，揭示Cx43协调生长和图案化伴随。为了确定Cx43如何介导这些活动，使用分子遗传学方法鉴定推定的靶基因。来自该靶基因列表的经验证的Cx43依赖性基因将被置于Cx43下游作用基因的有序途径中。因此，将有可能揭示在鳍的生长和图案化期间由Cx43调节的逐步分子途径。这一建议的智力价值在于，将揭示对骨骼形态和模式建立的机械见解。前50个靶基因的验证将在一个新的基于探究的本科生实验室课程中完成。因此，这一建议的更广泛的影响包括为大量学生提供科学实践的机会，同时还提供了一套发育生物学的核心实验室技能。智力的优点和更广泛的影响被完全整合在这个建议，既推进了骨骼形态发生的知识领域，并提供了如何在实验室环境中解决复杂的生物学问题的指导。

The vertebrate skeleton provides a variety of important functions including form, strength, flexibility, and protection of vital organs. Establishment of the complex form of the skeleton requires the coordination of growth with the differentiation of bone-forming cells and joint-forming cells. Indeed, much is known about how cells divide and about how relevant cell types differentiate. However, very little is understood about how these events are coordinated to give rise to skeletal elements of the correct size and shape. The gap junction protein Cx43 has been shown to regulate both cell proliferation and joint formation during zebrafish fin regeneration, revealing that Cx43 coordinates growth and patterning concomitantly. To determine how Cx43 mediates these activities, putative target genes were identified using a molecular genetics approach. Validated Cx43-dependent genes from this list of target genes will be placed in an ordered pathway of genes acting downstream of Cx43. Thus, it will be possible to reveal the step-wise molecular pathway(s) regulated by Cx43 during growth and patterning of the fin. The intellectual merit of this proposal is that mechanistic insights into the establishment of skeletal form and patterning will be revealed. Validation of the top 50 target genes will be completed in part during a new inquiry-based laboratory course for undergraduates. Thus, the broader impacts of this proposal include providing a large number of students access to the practice of science while also providing a set of core laboratory skills in Developmental Biology. The intellectual merit and the broader impacts are fully integrated in this proposal, both advancing the field of knowledge of skeletal morphogenesis and providing instruction on how to solve complex biological problems in a laboratory setting.