The role of spectraplakins as key integrators of axonal microtubule networks

Spectraplakins 作为轴突微管网络关键整合者的作用

• 批准号: BB/I002448/1
• 负责人: Andreas Prokop
• 金额: $ 54.91万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2011
• 资助国家: 英国
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FI002448%2F1
• 关键词: role; spectraplakins; key; integrators; axonal

The ability of neurons to extend long processes (axons) towards other neurons, muscles or glands, is a key process underlying the formation of the neuronal networks that make up our brain and coordinate our behaviour. For example, failure of axonal growth is often fatal or causes paralysis upon spinal injury. To find cures we have to acquire a thorough understanding of the mechanisms underlying axonal growth. Growing axons follow reproducible paths signposted by chemical cues that direct the growing axon. Axonal elongation is essentially driven by filamentous skeletal elements of cells, called actin and microtubules. Actin and microtubules have to cooperate closely, and their activity must be adaptable to instructions through the guiding chemical cues. Unravelling how these different factors (actin, microtubules and signals) integrate and cross-coordinate each other during axonal growth is the key task that needs to be addressed. To this end, we focus our work on putative integrator molecules, called spectraplakins. They can physically link microtubules to actin and signalling components and, in their absence, axonal growth is severely inhibited. However, how the links of spectraplakins to actin, microtubules, and signalling components help them to perform their growth promoting function remains to be resolved. To facilitate this task, we study the spectraplakin Short stop (Shot) of fruitflies, which represents a most helpful model. Thus, Shot can be studied with powerful and rapid strategies available in fruitflies; the data obtained are relevant and can be translated into biomedical research, since the characteristics and functions of Shot are virtually identical to those of mammalian or human spectraplakins. As the starting point of our project, we have identified three regions/domains of the Shot molecule that can link Shot to microtubules, actin and, most likely, signalling factors. We have shown that all of them are absolutely required for Shot function in axonal growth. Here we will address the detail of their function, using advanced microscopy, structure-function studies, and state-of-the art technology (mass spectrometry) to identify yet unknown binding factors. As an essential further strategy we will make systematic use of combinatorial genetics. We will combine mutations of Shot with mutations in other genes known to contribute to axonal growth. The combined mutant defects will give essential insights into the functional relationships of Shot to other factors, thus mapping Shot function into the systemic context of axonal growth. Our results will provide essential new insights into the function of spectraplakins in health and disease and the regulatory networks underlying axonal growth.

神经元将长突（轴突）延伸至其他神经元、肌肉或腺体的能力，是形成构成我们大脑并协调我们行为的神经元网络的关键过程。例如，轴突生长失败通常是致命的或在脊髓损伤时导致瘫痪。为了找到治疗方法，我们必须彻底了解轴突生长的机制。生长的轴突遵循由化学线索指示的可重复路径，这些化学线索指导着轴突的生长。轴突伸长本质上是由细胞的丝状骨骼元件（称为肌动蛋白和微管）驱动的。肌动蛋白和微管必须密切合作，并且它们的活性必须适应通过引导化学线索的指令。阐明这些不同因素（肌动蛋白、微管和信号）在轴突生长过程中如何相互整合和交叉协调是需要解决的关键任务。为此，我们将工作重点放在假定的积分分子上，称为“spectrumplakins”。它们可以将微管与肌动蛋白和信号成分物理连接，如果没有它们，轴突生长会受到严重抑制。然而，spectrumplakins 与肌动蛋白、微管和信号成分的联系如何帮助它们发挥促进生长的功能仍有待解决。为了促进这项任务，我们研究了果蝇的spectrumplakin Short stop（Shot），它代表了最有用的模型。因此，可以使用果蝇中强大而快速的策略来研究射击；所获得的数据具有相关性，可以转化为生物医学研究，因为 Shot 的特性和功能实际上与哺乳动物或人类的 Spectraplakins 相同。作为我们项目的起点，我们已经确定了 Shot 分子的三个区域/结构域，它们可以将 Shot 与微管、肌动蛋白以及最有可能的信号因子连接起来。我们已经证明，所有这些都是轴突生长中 Shot 功能所绝对必需的。在这里，我们将使用先进的显微镜、结构功能研究和最先进的技术（质谱）来详细介绍它们的功能，以识别尚不清楚的结合因子。作为一项重要的进一步策略，我们将系统地利用组合遗传学。我们将 Shot 的突变与已知有助于轴突生长的其他基因的突变结合起来。组合的突变缺陷将为 Shot 与其他因素的功能关系提供重要的见解，从而将 Shot 功能映射到轴突生长的系统背景中。我们的结果将为光谱蛋白在健康和疾病中的功能以及轴突生长的调控网络提供重要的新见解。

项目成果

Functional and Genetic Analysis of Spectraplakins in Drosophila.

• DOI: 10.1016/bs.mie.2015.06.022
• 发表时间: 2015-08
• 期刊: Methods in enzymology
• 作者: Ines Hahn;M. Ronshaugen;N. Sánchez-Soriano;A. Prokop

A new concept explaining the cell biology of axons and axon pathology

解释轴突细胞生物学和轴突病理学的新概念

• 发表时间: 2020
• 作者: Hahn, I.

A new concept explaining axonal cell biology, ageing and pathology

解释轴突细胞生物学、衰老和病理学的新概念

• 发表时间: 2021
• 作者: Hahn I

Spectraplakins promote microtubule-mediated axonal growth by functioning as structural microtubule-associated proteins and EB1-dependent +TIPs (tip interacting proteins).

• DOI: 10.1523/jneurosci.0416-12.2012
• 发表时间: 2012-07-04
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Alves-Silva J;Sánchez-Soriano N;Beaven R;Klein M;Parkin J;Millard TH;Bellen HJ;Venken KJ;Ballestrem C;Kammerer RA;Prokop A
• 通讯作者: Prokop A

Drosophila CLIP-190 and mammalian CLIP-170 display reduced microtubule plus end association in the nervous system.

果蝇夹190和哺乳动物夹170在神经系统中显示降低的微管和末端关联。

• DOI: 10.1091/mbc.e14-06-1083
• 发表时间: 2015-04-15
• 期刊: Molecular biology of the cell
• 影响因子: 3.3
• 作者: Beaven R;Dzhindzhev NS;Qu Y;Hahn I;Dajas-Bailador F;Ohkura H;Prokop A
• 通讯作者: Prokop A