Role of Lamellipodin in fast, clathrin-independent endocytosis.

Lamellipodin 在快速、不依赖网格蛋白的内吞作用中的作用。

• 批准号: BB/N000226/1
• 负责人: Matthias Krause
• 金额: $ 45.45万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FN000226%2F1
• 关键词: Role; Lamellipodin; fast; clathrin; independent

The human body is composed of billions of cells. In order for the body to function properly these cells have to communicate with each other. They communicate by sending out signals that can be recognized by "antenna", which are called "receptors", on the surface of other cells. When a signal reaches a receptor, the receptor becomes activated, which stimulates cells to react and change their behaviour. The signal is stopped by removal of the activated receptors from the cell surface by uptake into vesicles, small compartments that are surrounded by a membrane. This process is called "endocytosis". Endocytosis is important for the normal functioning of our body but if it "goes wrong" it can lead to disease. For example, in the case of wound healing, upon injury, cells in the wound send out signals to attract other cells to repair the wound. When these "repair" cells receive the signal, they actively migrate to the wound to close it. When the process of endocytosis is not working properly in the repair cells, wound healing may be delayed and this may contribute to many diseases such as diabetic foot ulcers, venous leg ulcers, and pressure ulceration (decubitus) of immobilised patients. Especially, in an ageing population pathological wound healing becomes a major challenge. Cells send out signals by filling small vesicles inside the cell "X" with the compound that acts as a signal. The vesicle is then transported to the cell membrane of cell "X", the boundary of the cell with the outside world, and the content is released to the outside when the membrane of the vesicle fuses with the membrane of the cell "X". Nerve cells have many contacts with each other, called synapses, to relay signals to each other. This is the fundamental basis of how the brain works. At these synapses the vesicles with the stored signal is released to activate a neighbouring nerve cell "Y" by fusion of the vesicle with the membrane of the nerve cell "X". The signal is then diffusing to nerve cell "Y" where it activates receptors. Upon release of the signal the vesicle membrane has to be retrieved in order for the vesicle to be made again. The retrieval of the membrane is mediated again by endocytosis and is called synaptic vesicle recycling. Aberrant synaptic vesicle recycling impairs the communication of the nerve cells and this is thought to contribute to many neuropsychiatric diseases, which are major challenges to life long health and wellbeing such as epilepsy, depression, and schizophrenia. We propose to study the basic mechanisms of endocytosis of receptors and synaptic vesicle recycling using single cells cultured in the laboratory. We will analyze how cells normally control the uptake of receptors using biochemical approaches. We will also visualize individual proteins recruited to the vesicles by labeling them with a fluorescent dye. This will allow us to analyze the recruitment in living cells in real time using a microscope. Our study might permit us to uncover the role of the different proteins for the uptake of the receptors thereby explaining the mechanism of this fundamental biological process. We will also be using a microscope to make movies of cells migrating in a dish while exposed to signals from other cells. We will use this assay to uncover how endocytosis may contribute to the directed movement of cells to the signal as it would normally happen during wound healing. The outcome of the project will give novel insights into the normal function of individual cells within our body and in the long run may provide the knowledge for the development of targeted therapies for diseases with altered endocytosis such as pathological wound healing as well as neuropsychiatric diseases.

人体由数十亿个细胞组成。为了使身体正常运作，这些细胞必须相互沟通。它们通过发出信号进行交流，这些信号可以被其他细胞表面的“天线”（称为“受体”）识别。当信号到达受体时，受体被激活，刺激细胞反应并改变其行为。信号通过从细胞表面移除活化的受体而被阻止，所述活化的受体通过摄取到囊泡（被膜包围的小隔室）中而被移除。这个过程被称为“内吞作用”。内吞作用对我们身体的正常运作很重要，但如果它“出错”，它可能导致疾病。例如，在伤口愈合的情况下，在受伤时，伤口中的细胞发出信号以吸引其他细胞来修复伤口。当这些“修复”细胞接收到信号时，它们会主动迁移到伤口以闭合伤口。当修复细胞中的内吞过程不能正常工作时，伤口愈合可能会延迟，这可能会导致许多疾病，如糖尿病足溃疡，静脉性腿部溃疡和固定患者的压力性溃疡（褥疮）。特别是在老龄化人群中，病理性伤口愈合成为一个主要挑战。细胞通过在细胞“X”内的小泡中填充作为信号的化合物来发出信号。然后，囊泡被运输到细胞“X”的细胞膜，即细胞与外界的边界，并且当囊泡的膜与细胞“X”的膜融合时，内容物被释放到外部。神经细胞之间有许多联系，称为突触，以相互传递信号。这是大脑如何工作的根本基础。在这些突触处，具有存储的信号的囊泡被释放，以通过囊泡与神经细胞“X”的膜的融合来激活相邻的神经细胞“Y”。然后信号扩散到神经细胞“Y”，在那里它激活受体。在释放信号时，囊泡膜必须被收回，以便再次制造囊泡。细胞膜的回收又由内吞作用介导，称为突触囊泡再循环。异常的突触囊泡再循环损害神经细胞的通信，这被认为是导致许多神经精神疾病的原因，这些疾病是终身健康和福祉的主要挑战，例如癫痫症，抑郁症和精神分裂症。 我们拟利用实验室培养的单细胞研究受体内吞和突触囊泡再循环的基本机制。我们将分析细胞通常如何使用生物化学方法控制受体的摄取。我们还将通过用荧光染料标记单个蛋白质来观察它们被招募到囊泡中。这将使我们能够使用显微镜在真实的时间内分析活细胞中的招募。我们的研究可能使我们能够揭示不同蛋白质对受体摄取的作用，从而解释这一基本生物过程的机制。我们还将使用显微镜拍摄细胞在培养皿中迁移的电影，同时暴露于来自其他细胞的信号。我们将使用这种测定来揭示内吞作用如何有助于细胞对信号的定向运动，因为它通常发生在伤口愈合期间。该项目的成果将为我们体内单个细胞的正常功能提供新的见解，从长远来看，可能为开发具有改变的内吞作用的疾病（如病理性伤口愈合以及神经精神疾病）的靶向治疗提供知识。

项目成果

Brain specific Lamellipodin knockout results in hyperactivity and increased anxiety of mice.

脑特异性层状蛋白敲除导致小鼠的多动症和焦虑增加。

• DOI: 10.1038/s41598-017-05043-3
• 发表时间: 2017-07-14
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Bodo C;Fernandes C;Krause M
• 通讯作者: Krause M

Glycogen synthase kinase 3 controls migration of the neural crest lineage in mouse and Xenopus.

• DOI: 10.1038/s41467-018-03512-5
• 发表时间: 2018-03-19
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Gonzalez Malagon SG;Lopez Muñoz AM;Doro D;Bolger TG;Poon E;Tucker ER;Adel Al-Lami H;Krause M;Phiel CJ;Chesler L;Liu KJ
• 通讯作者: Liu KJ

Nance-Horan Syndrome-like 1 protein negatively regulates Scar/WAVE-Arp2/3 activity and inhibits lamellipodia stability and cell migration

Nance-Horan 综合征样 1 蛋白负向调节 Scar/WAVE-Arp2/3 活性并抑制片状伪足稳定性和细胞迁移

• DOI: 10.1101/2020.05.11.083030
• 发表时间: 2020
• 作者: Law A

Lamellipodin promotes invasive 3D cancer cell migration via regulated interactions with Ena/VASP and SCAR/WAVE.

• DOI: 10.1038/onc.2016.47
• 发表时间: 2016-09-29
• 期刊: Oncogene
• 影响因子: 8

Dissection, Culture and Analysis of Primary Cranial Neural Crest Cells from Mouse for the Study of Neural Crest Cell Delamination and Migration.

小鼠原代颅神经嵴细胞的解剖、培养和分析，用于神经嵴细胞分层和迁移的研究。

• DOI: 10.3791/60051
• 发表时间: 2019
• 期刊: JoVE
• 作者: Gonzalez Malagon SG