Role of actin cytoskeleton regulators in craniofacial development and disease

肌动蛋白细胞骨架调节剂在颅面发育和疾病中的作用

基本信息

批准号：
8839951
负责人：
Shuyi Nie
金额：
$ 25.69万
依托单位：
GEORGIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8839951
关键词：
Accounting Actins Behavior Biochemical Biochemistry Biological Biological Models Candidate Disease Gene Cell Adhesion Molecules Cell Surface Extensions Cells Cellular Morphology Cellular biology Cephalic Congenital Abnormality Connective Tissue Cytoskeleton Defect Development DiGeorge Syndrome Disease Dorsal Electroporation Embryo Embryology Embryonic Development Employee Strikes Employment Etiology Face Genes Image Imaging Techniques Lead Light Mandibulofacial Dysostosis Mentors Microscopy Molecular Movement Myosin ATPase Neural Crest Neural Crest Cell Neural tube Phase Population Process Rana Regulation Regulator Genes Research Personnel Resolution Role Signal Pathway Skeleton Staging Structure Techniques Time Tissues Work base caldesmon cell behavior cell motility cellular imaging cleft lip and palate craniofacial in vivo interest loss of function migration molecular imaging research study sensory system tool

项目摘要

DESCRIPTION (provided by applicant): Neural crest is a unique population of cells that migrate extensively during vertebrate embryogenesis. Of particular interest, cranial neural crest (CNC) cells migrate throughout the face to form most of the craniofacial skeleton. Defects in their proliferation, migration, or survival lead to numerous diseases and birth defects, including cleft lip/palate, Treacher-Collins syndrome, and DiGeorge syndrome. Despite recent progress in understanding aspects of neural crest development, the mechanisms underlying neural crest migration are still unclear. Thus, studying their migratory behavior and the regulation of actin cytoskeleton therein is crucial for understanding the basis of congenital craniofacial disorders. From a microarray screen, I have identified multiple actin cytoskeleton regulator (ACR) genes that are predominantly expressed in neural crest cells. My preliminary studies reveal that Myosin-X and Caldesmon are both critical for proper CNC cell migration via regulating cell morphology and membrane protrusions. Based on these findings, I hypothesis that neural crest cells achieve their unique migratory ability through specific ACRs to rapidly and precisely control their cytoskeletal dynamics. In this proposal, I will investigate the mechanism of neural crest migration by further analyzing the activities of these ACRs, and establishing their physical and functional interactions with actin structures. Specifically, I will combine classical embryology techniques with modern cell biology, biochemistry, and in vivo microscopy to study the dynamic arrangements of actin cytoskeleton during CNC migration and examine how ACRs in the neural crest regulate this process. Experiments will be carried out in frog and chick, taking into account the advantages of both model systems: simple gene and tissue manipulation in frog, and high-resolution imaging and late stage electroporation in chick. The following specific aims will be performed: 1) Examine the dynamics of actin cytoskeleton during normal neural crest migration by real time imaging. 2) Determine the role of Myosin-X and Caldesmon in regulating actin dynamics during CNC migration. 3) Functionally characterize Cdc42ep1 and additional candidate genes identified in the microarray screen for their roles in neural crest development. 4) Determine functional and physical interactions between ACRs in CNC migration.

描述（由申请人提供）：神经rest是在脊椎动物胚胎发生过程中广泛迁移的独特细胞群。特别有趣的是，颅神经rest（CNC）细胞在整个面部迁移，形成大多数颅面骨骼。它们的扩散，迁移或生存的缺陷导致许多疾病和先天缺陷，包括唇裂/pa/palete，Treacher-Collins综合征和Digeorge综合征。尽管在理解神经rest发育方面的方面最近取得了进展，但仍不清楚神经rest迁移的机制。因此，研究其迁移行为和其中的肌动蛋白细胞骨架的调节对于理解先天性颅面疾病的基础至关重要。从微阵列筛选中，我已经确定了多个肌动蛋白细胞骨架调节剂（ACR）基因，这些基因主要表达在神经rest细胞中。我的初步研究表明，肌球蛋白X和Caldesmon对于通过调节细胞形态和膜突起而适当的CNC细胞迁移至关重要。基于这些发现，我假设神经rest细胞通过特定的ACR实现其独特的迁移能力，以快速而精确地控制它们的细胞骨架动力学。在此提案中，我将通过进一步分析这些ACR的活性并建立其与肌动蛋白结构的物理和功能相互作用来研究神经rest迁移的机制。具体而言，我将将经典的胚胎学技术与现代细胞生物学，生物化学和体内显微镜相结合，以研究CNC迁移过程中肌动蛋白细胞骨架的动态排列，并检查神经crest中的ACR在神经cn迁移中如何调节这一过程。考虑到实验将在青蛙和小鸡中进行两个模型系统的优点：青蛙中的简单基因和组织操纵，以及雏鸡的高分辨率成像和晚期电穿孔。将执行以下特定目的：1）检查正常神经波峰通过实时成像迁移期间肌动蛋白细胞骨架的动力学。 2）确定肌球蛋白X和Caldesmon在调节CNC迁移过程中调节肌动蛋白动力学中的作用。 3）功能表征CDC42EP1和在微阵列屏幕中鉴定出的其他候选基因在神经rest发育中的作用。 4）确定CNC迁移中ACR之间的功能和物理相互作用。