Characterizing the neural crest response to BMP signaling through gastrulation and neurulation

项目总结 颅神经脊(NC)有助于许多颅面结构的形成，包括 面部的骨和软骨、牙本质和外周神经节。细胞信号调节不同方面 颅骨NC规范、上皮向间充质转化(EMT)和分化及破坏 这一发育计划导致了许多颅骨NC衍生的头面部出生缺陷，包括 颅骨融合症、Treacher Collins和Charge综合征以及腭裂。BMP信号在其中起着至关重要的作用 在颅骨NC的规范和分化中的作用，以及最近，BMP信号被显示为 控制头颅数控EMT。骨形态发生蛋白信号在颅脑NC中作用的机制研究 发展是开发新的头面部疾病预防和治疗措施的关键 缺陷。 这一建议将确定鸡骨形成蛋白梯度形成的分子机制。 原肠胚，以及这种梯度如何调节脑细胞类型的形成，包括神经，颅骨NC， 胎盘和表皮命运。这些实验将使用活体和硅胶方法来测试 细胞外BMP配体主要由颅骨NC产生并主动穿梭的假说 远距离信号在非神经性外胚层最强。接下来，进行定量的表达式分析 并将使用实时成像来建立原肠形成和神经形成期间BMP信号的时间表， 对所得数据集的分析将确定人群和单细胞水平对BMP的反应 信号。信号定时和强度的差异将与对不同目标的直接输入相关 基因。最后，我们将研究BMP靶基因Id1/2/3/4和Fibin在颅脑NC EMT中的作用 使用体内功能分析。总而言之，这些目标的结果将提供一个全面的 了解BMP信号事件在早期颅骨NC发育中的调节和作用。 除了确定预防头面部出生缺陷的翻译途径的目标外， 该提案的指导阶段将为Michael Piacentino博士提供必要的培训，因为他准备 开始了他的独立生涯。加州理工学院的玛丽安·布朗纳博士的实验室和他的 组建顾问委员会，提供必要的工具、专业知识和培训环境，以有效地 执行提议的目标，确立皮亚肯蒂诺博士的独立性。这一培训将是非常有用的 皮亚肯蒂诺博士开始了他的独立研究计划，并将提供所需的经验 在颅面发育过程中对骨形态发生蛋白信号的持续影响。

PROJECT SUMMARY The cranial neural crest (NC) contributes to the formation of many craniofacial structures including the bones and cartilage of the face, tooth dentin and peripheral ganglia. Cell signaling regulates different aspects of cranial NC specification, epithelial-to-mesenchymal transition (EMT) and differentiation and disruptions in this developmental program results in many cranial NC-derived craniofacial birth defects including craniosynestosis, Treacher Collins and CHARGE syndromes, and cleft palate. BMP signaling plays a crucial role during the specification and differentiation of cranial NC, and more recently, BMP signaling was shown to control cranial NC EMT. A mechanistic understanding of the role of BMP signaling during cranial NC development is essential to develop novel preventative and therapeutic measures against craniofacial defects. This proposal will determine the molecular mechanism of BMP gradient formation in the chick gastrula, and how this gradient regulates the formation of cranial cell types including neural, cranial NC, placode and epidermal fates. These experiments will use in vivo and in silico approaches to test the hypothesis that extracellular BMP ligands are produced primarily by the cranial NC and are actively shuttled over long distances to signal most strongly in the nonneural ectoderm. Next, quantitative expression analysis and live imaging will be used to establish the timeline of BMP signaling during gastrulation and neurulation, and analysis of the resulting datasets will determine population- and single-cell-level responses to BMP signals. Differences in signal timing and strength will then be correlated with direct input into different target genes. Finally, the role of BMP target genes Id1/2/3/4 and Fibin during cranial NC EMT will be investigated using in vivo functional analyses. Together, the results of these aims will provide a comprehensive understanding of the regulation and roles of BMP signaling events during early cranial NC development. In addition to identifying targets for translational avenues to prevent craniofacial birth defects, the mentored phase of this proposal will provide Dr. Michael Piacentino with necessary training as he prepares to begin his independent career. Dr. Marianne Bronner's lab at California Institute of Technology, and his assembled advisory council, provide the necessary tools, expertise, and training environment to efficiently execute the proposed aims and establish Dr. Piacentino's independence. This training will be instrumental as Dr. Piacentino begins his independent research program and will provide the experience needed to make lasting impacts on the field of BMP signaling during craniofacial development.