F-BAR proteins in neuronal migration and process formation

F-BAR 蛋白在神经元迁移和过程形成中的作用

基本信息

批准号：
10317364
负责人：
Erik W Dent
金额：
$ 38.88万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10317364
关键词：
Actin-Binding Protein Actins Address Affect Axon Binding Brain CRISPR/Cas technology Cell membrane Cell physiology Cells Cerebral cortex Cytoskeleton Data Dendrites Destinations Development Electroporation Endocytic Vesicle Endocytosis Event Family Family member Filopodia Gap Junctions Huntington Disease Immunoprecipitation Label Link Malignant Neoplasms Mammalian Cell Mass Spectrum Analysis Membrane Microfilaments Microtubules Molecular Morphology Mus Nerve Degeneration Nervous System Physiology Neurites Neurons Pattern Peripheral Play Polymers Positioning Attribute Process Protein Family Proteins Publishing RNA Splicing Reagent Reporter Research Role Set protein Signal Pathway Structure TRIP10 gene Techniques Testing Variant Vesicle Work cell type design dimer dosage experimental study human disease in utero in vivo insight internal control knock-down migration nervous system development neuron development neuronal replacement novel novel strategies overexpression polymerization protein function rho GTP-Binding Proteins

项目摘要

Breaking neuronal symmetry is a fundamental process in the formation of a polarized neuron. Neurons in the developing cerebral cortex are born as spherical cells that must extend leading and trailing processes to migrate to their destination in the developing cortical plate. Cortical neurons then extend long axons and dendrites from these processes to create functional circuits. Cortical neuron migration and process extension is critically dependent on the microtubule and actin cytoskeleton, but relatively little is known about how the actin cytoskeleton and plasma membrane are coordinated during these events. Membrane protrusion and invagination are fundamental cellular activities that require coordination of the plasma membrane and underlying actin cytoskeleton. However, there is a dearth of data on how membrane protrusion and invagination are integrated in process outgrowth and neuronal migration. The F-BAR superfamily of proteins are involved in membrane curvature sensing and deformation through their F-BAR domain, positioning them as potentially important players in both membrane invagination and protrusion. Structurally, they form a curved dimer that self-multimerizes around endocytic vesicles, causing their elongation into tubules. The CIP4 family of proteins (CIP4, FBP17 and TOCA1) is a group of F-BAR proteins that bind actin-associated proteins. Like other F-BAR proteins, the CIP4 family is thought to function primarily in membrane invagination and endocytosis, but our recent work has implicated CIP4 in neuronal membrane protrusion as well. Lamellipodial-like protrusions induced by CIP4 strongly inhibit neurite outgrowth in culture. Conversely, we find that a close family member, FBP17, forms endocytic tubules in developing cortical neurons and promotes prominent filopodia formation, resulting in precocious neurite outgrowth. In this proposal we will test the novel hypothesis that protrusion through CIP4 and invagination through FBP17 act in opposing manners to regulate cortical neuron migration and process formation in the developing cortex. Specifically, we will: 1) Determine how CIP4 induces membrane protrusions and FBP17 forms endocytic tubules, 2) Establish how membrane tubulation results in precocious filopodia formation and neurite outgrowth, 3) Reveal the spatial and temporal expression pattern of endogenously-labeled CIP4 and FBP17 in mouse lines and 4) Resolve CIP4 and FBP17 function in cortical development in vivo. This work will provide fundamental insights into how proteins that bridge the membrane and actin cytoskeleton function to regulate process outgrowth and cortical neuron migration in the early developing mammalian brain. CIP4 and FBP17 have been implicated in Huntington's disease and several forms of cancer, underscoring the importance of determining the function of these proteins in the developing cerebral cortex.

打破神经元对称性是极化神经元形成的基本过程。神经元在发育中的大脑皮层天生是球形细胞，必须延伸前导过程和尾随过程才能迁移到达正在发育的皮质板中的目的地。然后皮质神经元从这些过程创建功能电路。皮质神经元迁移和过程扩展至关重要依赖于微管和肌动蛋白细胞骨架，但对于肌动蛋白细胞骨架如何和质膜在这些事件期间协调。膜突出和内陷是需要质膜和底层肌动蛋白细胞骨架协调的基本细胞活动。然而，缺乏关于膜突出和内陷如何在过程中整合的数据生长和神经元迁移。 F-BAR 蛋白质超家族参与膜曲率传感和通过其 F-BAR 域变形，将它们定位为两种膜中潜在的重要参与者内陷和突出。在结构上，它们形成弯曲的二聚体，围绕内吞进行自多聚化囊泡，导致它们伸长成小管。 CIP4 蛋白家族（CIP4、FBP17 和 TOCA1）是一组结合肌动蛋白相关蛋白的 F-BAR 蛋白。与其他 F-BAR 蛋白一样，CIP4 家族被认为主要在膜内陷和内吞作用中发挥作用，但我们最近的工作表明 CIP4 在神经元中发挥作用膜突出也。 CIP4诱导的片状伪足样突起强烈抑制神经突的生长文化。相反，我们发现一个亲密的家族成员 FBP17 在发育中的皮质中形成内吞小管神经元并促进显着的丝状伪足形成，导致神经突过早生长。在这个提案中我们将测试新的假设，即通过 CIP4 的突出和通过 FBP17 的内陷起到相反的作用调节发育中皮层中皮层神经元迁移和过程形成的方式。具体来说，我们将： 1) 确定 CIP4 如何诱导膜突出和 FBP17 形成内吞小管，2) 确定如何膜管导致早熟丝状伪足形成和神经突生长，3）揭示空间和小鼠品系中内源标记的 CIP4 和 FBP17 的时间表达模式以及 4) 解析 CIP4 和 FBP17 在体内皮质发育中发挥作用。这项工作将为蛋白质如何连接膜和肌动蛋白细胞骨架功能，调节过程生长和皮质神经元迁移在早期发育的哺乳动物大脑中。 CIP4 和 FBP17 与亨廷顿舞蹈病有关多种形式的癌症，强调确定这些蛋白质在发育中的功能的重要性大脑皮层。