Control of AMPA receptor trafficking during hippocampal long-term depression (LTD): the role of the p38 MAPK-MK2-LIMK1-cofilin1 signalling cascade

海马长期抑郁 (LTD) 期间 AMPA 受体运输的控制：p38 MAPK-MK2-LIMK1-cofilin1 信号级联的作用

• 批准号: BB/H018344/1
• 负责人: Sonia Correa-Muller
• 金额: $ 46.9万
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2010
• 资助国家: 英国
• 起止时间: 2010 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FH018344%2F1
• 关键词: Control; AMPA; receptor; trafficking; during

Learning and memory are processes associated with long-lasting change in information flow between nerve cells in the brain (synapse transmission). Decreased information flow, known as long-term depression (LTD), is induced in the hippocampus by the activation of group I-metabotropic glutamate receptors (GpI-mGluR), which leads through a cascade of events to the loss of AMPA receptor (AMPAR) from the synapse. Therefore, it is very important to know the identity and functional role of the proteins that are involved in the cascade of events initiated by the activation of GpI-mGluR located on the surface of the nerve cell leading to the loss (internalisation) of AMPAR also situated on the surface of the nerve cell. This process involves several steps and different proteins. p38 mitogen-activated protein kinase (p38 MAPK) is known to be involved in the induction of GpI-mGluR-LTD. A kinase is an enzyme which function is to add phosphate to another protein (substrate). The added phosphate activates or inhibits the function of the substrate. During GpI-mGluR-LTD, the substrates for p38 MAPK are not yet known. The aim of this proposal is to test the hypothesis that MK2, which is a direct substrate of p38 MAPK, is the missing link between mGluR-mediated p38 MAPK activation and the internalisation of the AMPAR during mGluR-LTD using a broad range of techniques. Pilot data have already found cofilin1 as a potential target of p38 MAPK during GpI-mGluR-LTD. Furthermore, increased levels in p38 MAPK and MK2 phosphorylation was also observed in mGluR-LTD. To confirm the biological function of p38 MAPK-MK2-LIMK1-cofilin1 signalling cascade, we will use primary cultures from MK2/3 and MK2 knockout mice combined with molecular biology techniques, immunocytochemistry, confocal imaging and electrophysiological recordings. First, we will use neurons lacking MK2/3 and induce mGluR-LTD to address the internalisation of AMPAR. We will also replace the phosphorylated amino acids in the cofilin1 and LIMK1 to inactive amino-acids and determine whether the replacement affects the function of the endogenous protein. For example, if the endogenous protein is involved in the internalisation of AMPAR, will the manipulated protein block the internalisation of AMPAR. AMPAR has a very important role in neuronal function and therefore in brain function. Loss of brain function, which can be caused by several factors including normal ageing, has been associated with several neurological disorders such as Alzheimer's disease. Thus, understanding the mechanisms regulating internalisation of AMPAR will provide insight into both normal and abnormal synaptic mechanisms and may also identify novel targets to slow cognitive decline in ageing and in neurodegeneration.

学习和记忆是与大脑中神经细胞之间的信息流（突触传递）的长期变化相关的过程。减少的信息流，被称为长期抑郁症（LTD），是由I组代谢型谷氨酸受体（GpI-mGluR）的激活在海马中诱导的，这通过一系列事件导致AMPA受体（AMPAR）从突触中丢失。因此，了解参与由位于神经细胞表面上的GpI-mGluR的激活引发的级联事件的蛋白质的身份和功能作用非常重要，所述事件导致也位于神经细胞表面上的AMPAR的损失（内化）。这个过程涉及几个步骤和不同的蛋白质。已知p38丝裂原活化蛋白激酶（p38 MAPK）参与GpI-mGluR-LTD的诱导。激酶是一种酶，其功能是将磷酸盐添加到另一种蛋白质（底物）上。添加的磷酸盐激活或抑制底物的功能。在GpI-mGluR-LTD期间，p38 MAPK的底物尚不清楚。该提案的目的是测试的假设，MK2，这是一个直接底物的p38 MAPK，是mGluR介导的p38 MAPK激活和内化的AMPAR在mGluR-LTD使用广泛的技术之间的缺失环节。为了证实p38 MAPK-MK2-LIMK 1-cofilin 1信号通路的生物学功能，我们将使用MK2/3和MK2基因敲除小鼠的原代培养物，结合分子生物学技术，免疫细胞化学，共聚焦成像和电生理记录。首先，我们将使用缺乏MK 2/3的神经元并诱导mGluR-LTD来解决AMPAR的内化。我们还将替换cofilin 1和LIMK 1中的磷酸化氨基酸为无活性氨基酸，并确定替换是否影响内源性蛋白质的功能。例如，如果内源性蛋白质参与AMPAR的内化，那么被操纵的蛋白质是否会阻断AMPAR的内化。AMPAR在神经元功能中具有非常重要的作用，因此在脑功能中也具有非常重要的作用。脑功能的丧失可能由包括正常衰老在内的多种因素引起，与阿尔茨海默病等多种神经系统疾病有关。因此，了解AMPAR的内化调节机制将提供对正常和异常突触机制的深入了解，并且还可以确定减缓衰老和神经变性中认知下降的新靶点。

项目成果

Activity-Regulated Cytoskeleton-Associated Protein Controls AMPAR Endocytosis through a Direct Interaction with Clathrin-Adaptor Protein 2.

• DOI: 10.1523/eneuro.0144-15.2016
• 发表时间: 2016-05
• 期刊: eNeuro
• 影响因子: 3.4
• 作者: DaSilva LL;Wall MJ;P de Almeida L;Wauters SC;Januário YC;Müller J;Corrêa SA
• 通讯作者: Corrêa SA

The mechanistic link between Arc/Arg3.1 expression and AMPA receptor endocytosis.

• DOI: 10.1016/j.semcdb.2017.09.005
• 发表时间: 2017-09
• 期刊: Seminars in cell & developmental biology
• 影响因子: 7.3
• 作者: Mark J. Wall;Sonia A. L. Corrêa

The MK2/3 cascade regulates AMPAR trafficking and cognitive flexibility.

• DOI: 10.1038/ncomms5701
• 发表时间: 2014-08-19
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Eales, Katherine L.;Palygin, Oleg;O'Loughlin, Thomas;Rasooli-Nejad, Seyed;Gaestel, Matthias;Mueller, Juergen;Collins, Dawn R.;Pankratov, Yuriy;Correa, Sonia A. L.
• 通讯作者: Correa, Sonia A. L.

The Temporal Dynamics of Arc Expression Regulate Cognitive Flexibility.

• DOI: 10.1016/j.neuron.2018.05.012
• 发表时间: 2018-06-27
• 期刊: Neuron
• 影响因子: 16.2
• 作者: Wall MJ;Collins DR;Chery SL;Allen ZD;Pastuzyn ED;George AJ;Nikolova VD;Moy SS;Philpot BD;Shepherd JD;Müller J;Ehlers MD;Mabb AM;Corrêa SAL
• 通讯作者: Corrêa SAL

The Role of p38 MAPK and Its Substrates in Neuronal Plasticity and Neurodegenerative Disease.

• DOI: 10.1155/2012/649079
• 发表时间: 2012
• 期刊: Journal of signal transduction
• 作者: Corrêa SA;Eales KL
• 通讯作者: Eales KL