Tissue-specific regulation of cellular morphogenesis during embryonic development of the invertebrate chordate Ciona intestinalis

 描述(申请人提供)：我对对胚胎发育有指导意义的基因调控和细胞过程有广泛的兴趣。作为一名研究生，我在被囊状肠囊的胚胎神经和心脏前体细胞的规格、构型和形态发生方面有了几项发现。意识到乔纳和其他被囊类动物作为发育遗传学模式生物的潜力很大程度上尚未开发，我继续进行这些无脊椎动物的博士后研究工作。在我的博士后期间，我开发了更多的工具来研究Ciona的基因功能，并确立了我作为被囊研究社区的年轻领导者的地位。K99/R00奖将允许我完成我的正式培训，并开始过渡到作为一名独立研究员的角色，领导一个研究被囊胚胎发育中细胞形态发生的研究小组。在这个项目中，我打算研究基因在调节Ciona胚胎中两种不同类型的细胞--躯干腹侧细胞(TVCs)和双极尾部神经元(BTN)--迁移和极化中的功能。在K99指导阶段，我将首先通过CRISPR/Cas9介导的突变在TVCs中研究基因功能，TVCs经历了刻板的定向迁移和极性定向。然后，我将描述TE BTN的转录组，识别可能调节刻板印象的定向迁移和极性取向的候选基因。这些目标的完成将为我实现我的第三个目标做好准备，这将在R00指导阶段执行，测试BTN中对这些候选基因的要求，允许在同一胚胎的不同壁龛中不同细胞类型的迁移和极性的分子控制之间进行直接比较。指导阶段符合我的导师克里斯蒂安博士和我的研究兴趣。我们的目标都是了解细胞行为是如何受到调控的，以及这种调控的缺陷如何导致发育综合征和人类疾病。我希望在独立阶段及以后将自己与导师区分开来，这符合我对神经发育和神经元分化的特殊兴趣，而克里斯蒂安博士则专注于心脏发育。K99奖将允许我通过学习和有意义地应用FAC、RNA-SEQ和自动成像技术来完成我的博士后培训。K99还将使我能够生成我可以随身携带的初步数据，作为独立研究计划的基础。

 DESCRIPTION (provided by applicant): I am broadly interested in gene regulation and cellular processes that are instructive for embryonic development. As a graduate student, I made several discoveries relating to the specification, patterning, and morphogenesis of embryonic neural and cardiac progenitors in the tunicate Ciona intestinalis. Recognizing the largely untapped potential of Ciona and other tunicates as model organisms for developmental genetics, I have continued my postdoctoral research working on these invertebrate chordates. During my post-doc, I have developed additional tools for the study of gene function in Ciona and established myself as a young leader of the tunicate research community. The K99/R00 award will allow me to complete my formal training and begin my transition to a role as an independent investigator, at the helm of a research group studying cellular morphogenesis in tunicate embryonic development. For this project, I propose to study gene function in regulation of migration and polarization of two distinct cell types in the Ciona embryo, the Trunk Ventral Cells (TVCs) and the Bipolar Tail Neurons (BTNs). In the K99 mentored phase, I will first investigate gene function by CRISPR/Cas9- mediated mutagenesis in the TVCs, which undergo stereotyped directed migration and polarity orientation. I will then profile the transcriptome of te BTNs, identifying Candidate genes that may regulate stereotyped directed migration and orientation of polarity. The completion of these aims will prepare me for pursuing my third aim, to be performed in the R00 mentored phase, of testing the requirement for those Candidate genes in the BTNs, allowing for direct comparisons between the molecular control of migration and polarity in different cell types in different niches of the same embryo. The mentored phase fits the research interests of both my mentor, Dr. Christiaen, and I. We both aim to understand how cell behavior is regulated, and how deficiencies in this regulatory control may result in developmental syndromes and human disease. Where I hope to distinguish myself from my mentor during the independent phase and beyond is in my particular interest in neural development and neuronal differentiation, in contrast to Dr. Christiaen's focus on cardiac development. The K99 award will allow me to complete my postdoctoral training, by learning and meaningfully applying FACS, RNA-seq, and automated imaging techniques. The K99 will also enable me to generate preliminary data that I can take with me as the foundation for an independent research program.