Regulation of Axial Extension in Vertebrate Embryos

脊椎动物胚胎轴向延伸的调节

基本信息

批准号：
8699787
负责人：
Paul Michael Skoglund
金额：
$ 29.84万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8699787
关键词：
Actins Acute Adhesions Anencephaly Anterior Automobile Driving Biological Biological Assay Biomechanics Blastopores Cadherins Cell Shape Cell-Cell Adhesion Cells Chemicals Complex Congenital Abnormality Crosslinker Data Defect Dependence Development Diagnostic Dorsal Embryo Equilibrium Event Failure Feedback Filament Germ Layers Goals Health Homeostasis Human Infertility Intercalated Cell Lead Lip structure MYLK gene Mediating Mesoderm Cell Modeling Molecular Morphogenesis Motor Myosin ATPase Myosin Regulatory Light Chains Myosin Type II Neural Tube Closure Neurons Nonmuscle Myosin Type IIB Pathway interactions Pattern Pharmaceutical Preparations Phase Phosphorylation Phosphotransferases Probability Process Production Proteins Rana Regulation Research Personnel Role Series Shapes Signal Transduction Spinal Dysraphism Stress Stretching System Takeda brand of pioglitazone hydrochloride Testing Therapeutic Intervention Tissues Up-Regulation Western Blotting Work base crosslink gastrulation inhibitor/antagonist intercalation metaplastic cell transformation myosin VI myosin phosphatase non-muscle myosin novel relating to nervous system research study

Actins Acute Adhesions Anencephaly Anterior Automobile Driving Biological Biological Assay Biomechanics Blastopores Cadherins Cell Shape Cell-Cell Adhesion Cells Chemicals Complex Congenital Abnormality Crosslinker Data Defect Dependence Development Diagnostic Dorsal Embryo Equilibrium Event Failure Feedback Filament Germ Layers Goals Health Homeostasis Human Infertility Intercalated Cell Lead Lip structure MYLK gene Mediating Mesoderm Cell Modeling Molecular Morphogenesis Motor Myosin ATPase Myosin Regulatory Light Chains Myosin Type II Neural Tube Closure Neurons Nonmuscle Myosin Type IIB Pathway interactions Pattern Pharmaceutical Preparations Phase Phosphorylation Phosphotransferases Probability Process Production Proteins Rana Regulation Research Personnel Role Series Shapes Signal Transduction Spinal Dysraphism Stress Stretching System Takeda brand of pioglitazone hydrochloride Testing Therapeutic Intervention Tissues Up-Regulation Western Blotting Work base crosslink gastrulation inhibitor/antagonist intercalation metaplastic cell transformation myosin VI myosin phosphatase non-muscle myosin novel relating to nervous system research study

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项目摘要

DESCRIPTION (provided by applicant): The proposal "Regulation of Vertebrate Axial Extension" aims to expand on the observation previously made by this investigator that a cortical acto-myosin network is required for the cellular convergence and extension that drives the shape change of frog embryos at gastrulation. Because this sub-membranous actin network requires the myosin IIB (MIIB) complex to function in this morphogenesis we aim to determine whether this MII dependence requires actin-activated contractility, or if MII functions solely as a actin crosslinker during axial extension. Preliminary evidence depleting the myosin regulatory light chain (MRLC), required for expression of contractile but not crosslinking activity in MII, indicates that myosin II contractility is required. Because regulation of MII contractility is controlled by the phosphorylation state of Ser19 on MRLC, we perturbed a possible dynamic homeostasis of this Ser19-P event by adding an inhibitor of myosin phosphatase to embryos, and assaying for Ser19 phosphorylation by quantitative western blotting to reveal an acute upregulation of Ser19-P. This experiment reveals that there is an active, balanced process involving myosin phosphatase and a kinase or kinases to regulate Ser19-P levels in intercalating cells, and further acute drug experiments indicate that the relevant kinase is MLCK. Because Ca ++ is an upstream regulator of MLCK, we reexamined Ca++ dynamics in intercalating cells, finding a novel pattern of Ca++ fluxes, as well as a plausible mechanism for generating these fluxes through a putative Ca++ channel protein that we show to be required for gastrulation. We also identify a role for a second motor protein in regulating the cortical actin network, allowing us to propose a model for CE consisting of a simple oscillating regulatory circuit with biomechanical feedback control, as well as the means to test this hypothesis.

描述（由申请人提供）：提案“脊椎动物轴向延伸的调节”旨在扩展该研究者先前对的观察结果，即细胞收敛和延伸的皮质肌动肌动蛋白网络是必需的，该网络需要驱动胃肠道的青蛙胚胎的形状变化。由于该亚膜肌动蛋白网络需要肌球蛋白IIB（MIIB）复合物在这种形态发生中起作用，因此我们旨在确定该MII依赖性是否需要肌动蛋白激活的收缩力，或者MII在轴向扩展过程中仅充当肌动蛋白交叉链接。肌球蛋白调节轻型链（MRLC）的初步证据表达收缩性而不是MII中的交联活动所需的，这表明需要肌球蛋白II II收缩性。 Because regulation of MII contractility is controlled by the phosphorylation state of Ser19 on MRLC, we perturbed a possible dynamic homeostasis of this Ser19-P event by adding an inhibitor of myosin phosphatase to embryos, and assaying for Ser19 phosphorylation by quantitative western blotting to reveal an acute upregulation of Ser19-P.该实验表明，存在一个涉及肌球蛋白磷酸酶的活性，平衡的过程，以及一种激酶或激酶，以调节插入细胞中的Ser19-P水平，进一步的急性药物实验表明相关激酶是MLCK。由于Ca ++是MLCK的上游调节剂，因此我们在插入细胞中重新检查了Ca ++动力学，找到了Ca ++通量的新模式，以及通过推定的Ca ++通道蛋白产生这些通量的合理机制，我们证明我们是胃肠道所必需的。我们还确定了第二个运动蛋白在调节皮质肌动蛋白网络中的作用，使我们能够为CE提出一个模型，该模型由具有生物力学反馈控制的简单振荡调节回路以及检验该假设的手段。