Regulation of embryonic epithelial morphogenesis

胚胎上皮形态发生的调节

• 批准号: 7898061
• 负责人: Shaohua Li
• 金额: $ 10.86万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7898061
• 关键词: 1-Phosphatidylinositol 3-Kinase; Adhesions; Affect; Apical; Apoptosis; Architecture; Basement membrane; Binding; Biochemical Genetics; Biological Models; Biology; Cancer Biology; Cell Polarity; Cell Survival; Cell membrane; Cells; Complex; Cues; DNA; DOCK1 protein; Data; Defect; Development; Dominant-Negative Mutation; Embryo; Embryonic Development; Environment; Epiblast; Epithelial; Epithelial cyst; Epithelium; Event; Exhibits; Exocytosis; Experimental Models; Figs - dietary; GTP Binding; Genes; Goals; Guanine Nucleotide Exchange Factors; IQ motif containing GTPase activating protein 1; In Situ Nick-End Labeling; In Vitro; Inner Cell Mass; Integrins; Lead; Maintenance; Malignant - descriptor; Mediating; Methods; Microtubules; Molecular; Morphogenesis; Mus; Natural regeneration; Pathway interactions; Phenotype; Play; Process; Proteins; Recruitment Activity; Regulation; Retroviridae; Role; Signal Transduction; Staging; Staining method; Stains; Testing; Time; Tissue Engineering; Tissues; Undifferentiated; Work; Wound Healing; blastocyst; caspase-3; embryonic stem cell; extracellular; genetic manipulation; implantation; integrin-linked kinase; knock-down; mutant; natural Blastocyst Implantation; novel; protein complex; protein transport; rho GTP-Binding Proteins; small hairpin RNA; tissue regeneration; tool; trafficking

Formation of polarized epithelium is fundamental to embryonic development. During mouse peri- implantation development, the undifferentiated inner cell mass of the blastocyst is converted from a nonpolar cell aggregate to a highly organized epithelial cyst. This morphogenetic transformation involves the polarization and basement membrane (BM)-dependent survival of epiblast epithelium. However, the molecular mechanisms underlying these processes are largely unknown. The mouse peri-implantation development can be recapitulated by in vitro cultured embryoid bodies (EBs) differentiated from embryonic stem cells. Using genetically modified EBs, we show for the first time that BM formation directs the assembly of an adhesion complex, which serves as a signaling platform to regulate epiblast polarization and survival. Our preliminary data supporting this idea, demonstrating that (1) assembly of the BM-associated adhesion complexes correlates with the activation of Rho GTPases Cdc42 and Rac1; (2) Cdc42 controls microtubule organization and the trafficking of apical polarity proteins, processes that are required for cell elongation and polarization; and (3) Rac1 activation plays an essential role in BM-dependent epiblast survival. These data strongly support our hypothesis that BM formation induces epiblast polarization and promotes epiblast survival via activation of Cdc42 and Rac1. To test this hypothesis, we propose to determine the mechanism through which the assembly of BM-associated adhesion complexes induces Cdc42 and Rac1 activation (Specific Aim 1 and 3). Next, we will determine the Cdc42-effector interactions that regulate epiblast elongation and apical polarization, focusing on the role of IQGAP1 in microtubule capture and the exocyst in apical polarity protein trafficking (Specific Aim 2). Finally, we will determine the Rac1- effector interactions that regulates epiblast survival (Specific Aim 4). The long-term goal of this study is to identify the extracellular cues and the transmembrane cascades that regulate embryonic epithelial morphogenesis, which are crucial to our understanding of epithelial biology, embryogenesis, and tissue regeneration. Elucidating the molecular mechanisms of embryonic epithelial tissue formation that integrate cellular polarity, differentiation and survival with tissue architecture is not only critical to our understanding of embryonic development and cancer biology but also has implications in tissue regeneration and tissue engineering.

极化上皮的形成是胚胎发育的基础。在小鼠交配期间- 在着床发育过程中，胚泡的未分化内细胞团从 非极性细胞聚集成高度组织化的上皮囊肿。这种形态发生的转变 涉及上胚层上皮的极化和基底膜（BM）依赖性存活。 然而，这些过程背后的分子机制在很大程度上是未知的。 体外培养的拟胚体可再现小鼠围着床期的发育过程 (EBs)从胚胎干细胞分化而来。使用转基因EB，我们首先展示了 BM形成指导粘附复合物的组装的时间，粘附复合物用作信号传导。 调节外胚层极化和存活的平台。我们的初步数据支持这一观点， 证明（1）BM相关粘附复合物的组装与 Rho GTP酶Cdc 42和Rac 1的激活;（2）Cdc 42控制微管组织， 顶端极性蛋白的运输，细胞伸长和极化所需的过程; （3）Rac 1的激活在BM依赖的上胚层存活中起重要作用。这些数据强烈 支持我们的假设，即BM形成诱导上胚层极化并促进上胚层存活 通过激活Cdc 42和Rac 1。为了验证这一假设，我们建议确定机制 BM相关粘附复合物的组装诱导Cdc 42和Rac 1活化 （具体目标1和3）。接下来，我们将确定Cdc 42-效应物相互作用，调节外胚层 伸长和顶端极化，重点是IQGAP 1在微管捕获和微管断裂中的作用。 顶极性蛋白运输中的外囊（特异性目的2）。最后，我们将确定Rac 1- 调节上胚层存活的效应物相互作用（特异性目的4）。 本研究的长期目标是确定细胞外信号和跨膜级联 调节胚胎上皮形态发生，这对于我们理解上皮细胞至关重要 生物学、胚胎发生和组织再生。阐明胚胎上皮组织形成的分子机制， 极性，分化和生存与组织结构不仅是至关重要的， 胚胎发育和癌症生物学，而且还涉及组织再生和组织 工程.