Dysregulation of Cav1.2 by beta amyloid peptide

β 淀粉样肽导致 Cav1.2 失调

• 批准号: 10521735
• 负责人: JOHANNES W HELL
• 金额: $ 161.05万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10521735
• 关键词: Adenylate Cyclase; Adrenergic Receptor; Affect; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid beta-Protein; Antibodies; Binding; Biotinylation; Brain; C-terminal; Chronic; Clinical; Complex; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Dendritic Spines; Development; Endocytosis; Endosomes; Etiology; Event; Excision; Excitatory Postsynaptic Potentials; Functional disorder; Gene Expression; Genetic Transcription; Glutamates; Hippocampus (Brain); Image; Impairment; Intervention; Isradipine; Knock-in; Knock-in Mouse; Label; Left; Link; Long-Term Depression; Long-Term Potentiation; Measures; Mediating; Mus; Neuronal Dysfunction; Neurons; Norepinephrine; Pathway interactions; Peptides; Permeability; Pharmacology; Phase; Phosphorylation; Phosphorylation Site; Proteins; Rattus; Recycling; Regulation; Regulator Genes; Reporting; Rodent; Rodent Model; Role; Science; Secretory Vesicles; Serine; Signal Pathway; Signal Transduction; Site; Surface; Symptoms; Synapses; Synaptic Transmission; Synaptic plasticity; Testing; Toxic effect; Up-Regulation; Western Blotting; abeta accumulation; aged; amyloid peptide; density; improved; inhibitor; innovation; mouse model; neuron loss; neuronal excitability; neurotoxic; neurotoxicity; peptide L; postsynaptic; protein kinase A kinase; recruit; reuptake; synaptic function; trafficking; uptake

Abstract Dysregulation of Cav1.2 by beta amyloid peptide L-type Ca2+ channels (LTCC) are key regulators of gene transcription, neuronal excitability, and synaptic functions. Cav1.2 is the prevalent LTCC in brain (Hell et al., 1993, JCB 123, 949-962). Chronically increased Ca2+ influx via LTCCs has been implicated early on in senile symptoms and Alzheimer’s disease (AD) (e.g., Science 272, 1017). We found that aged rats have significantly increased PKA-mediated phosphorylation of Cav1.2 on Serine 1928 (Davare and Hell, 2003, PNAS 100, 16018-23), which increases Cav1.2 channel activity (Qian, …, Hell, 2017, Sci Sig 10, eaaf9659). We also found that Cav1.2 forms a unique signaling complex with the b2 adrenergic receptor (b2 AR) that also contains all other proteins necessary for the cAMP-mediated regulation of Cav1.2 (e.g., Davare et al., 2001, Science 293, 98), making Cav1.2 a prime target for b2 AR signaling. b amyloid peptide 1-42 (Ab) is a hallmark of AD. Oligomeric Abo stimulates the b2 AR. Supportive evidence from single channel recording indicates that Abo increase Cav1.2 activity via the b2 AR. We hypothesize that this increase is mediated by phosphorylation of S1928, the main PKA site of Cav1.2, and triggers downstream events that ultimately cause neuronal damage. Aim 1 is to test whether Cav1.2 dysregulation by Abo via Cav1.2- associated b2 AR occurs in dendrites and spines (Ca2+ imaging), whether it is in part mediated by increased surface insertion of Cav1.2, and whether blocking this signaling pharmacologically or in innovative S1928A knock-in (KI) mice alleviates Abo neurotoxicity. Aim 2 is to test whether Abo - b2 AR - S1928 signaling stimulates surface insertion of Ca2+ permeable (CP) AMPARs (GluA1 homomers) and blocking CP-AMPARs alleviates Abo neurotoxicity. We will use cutting edge live imaging of SEP-tagged GluA1 and GluA2 and analyze field EPSPs and mEPSC before and after inhibition of glutamate re-uptake to measure AMPAR activity in the perisynaptic space. Aim 3 will test the role of Abo - b2 AR - S1928 signaling in augmentation of long-term depression by Abo, and in long-term potentiation, which is impaired by Abo. Crossing our S1928A KI mice with highly amyloidogenic 5xFAD mice will show whether Abo - b2 AR - S1928 signaling is important for Ab-induced dysfunction of Cav1.2 activity and synaptic transmission and its plasticity. This project is focused on what we hypothesize is an early effect of Abo (minute range) that triggers subsequent neurotoxic events because we need to understand all aspects of Abo toxicity. Also, as an early event the Abo - b2 AR - S1928 signaling constitutes a potential target for early pharmacological intervention in AD. We will to some degree explore later events downstream of Cav1.2 dysregulation by testing whether neurotoxicity in the 6-48h range can be improved by blockers of Abo - b2AR - CaV1.2 signaling and of CP-AMPAR in neuronal cultures. We will also explore whether blocking this signaling in the 5xFAD mouse model of AD will improve synapse function in these mice. The development of our peptide that disrupts binding of the b2 AR to Cav1.2 is highly innovative because it allows to specifically block signaling by Abo via the b2 AR without affecting other signaling pathways that involve the b2 AR.

摘要 β淀粉样肽对Cav1.2的调节异常 L型钙离子通道（LTCC）是基因转录、神经元兴奋性和突触传递的关键调节因子。 功能协调发展的Cav1.2是脑中普遍存在的LTCC（Hell等人，1993，JCB 123，949-962）。慢性增加 通过LTCC的Ca 2+内流在老年症状和阿尔茨海默病（AD）（例如， Science 272，1017）。我们发现老年大鼠PKA介导的磷酸化水平显著增加， Cav1.2作用于丝氨酸1928（Davare和Hell，2003，PNAS 100，16018-23），其增加Cav1.2通道活性 (Qian，Hell，2017，Sci Sig 10，eaaf 9659）。我们还发现Cav1.2形成了一种独特的信号复合物， b2肾上腺素能受体（b2 AR），还含有cAMP介导的所有其他蛋白质 Cav1.2的调节（例如，Davare等人，2001，Science 293，98），使得Cav1.2成为b2 AR信号传导的主要靶标。 B淀粉样肽1-42（Ab）是AD的标志。寡聚体ABO刺激b2 AR。支持性证据来自 单通道记录表明Abo通过b2 AR增加Cav1.2活性。我们假设这 增加是由S1928的磷酸化介导的，S1928是Cav1.2的主要PKA位点，并触发下游事件 最终导致神经元损伤目的1是测试是否Cav1.2失调由ABO通过Cav1.2- 相关的b2 AR发生在树突和棘（Ca 2+成像），无论它是部分介导的增加， Cav1.2的表面插入，以及是否阻止这种信号传导或在创新的S1928 A 基因敲入（KI）小鼠证实了Abo神经毒性。目的2是测试Abo-b2 AR - S1928信号传导是否刺激Abo-b2 AR - S1928信号传导。 Ca 2+渗透性（CP）AMPAR（GluA 1同聚体）的表面插入和阻断CP-AMPAR使Abo 神经毒性我们将使用SEP标记的GluA 1和GluA 2的尖端实时成像，并分析现场EPSP 和mEPSC在抑制谷氨酸再摄取之前和之后，以测量突触周膜中的AMPAR活性。 空间目的3将测试Abo-b2 AR-S1928信号传导在Abo增强长期抑郁中的作用， 和长时程增强，这是由ABO受损。将我们的S1928 A KI小鼠与高淀粉样蛋白生成小鼠杂交 5xFAD小鼠将显示Abo-b2 AR-S1928信号传导对于Ab诱导的Cav1.2功能障碍是否重要。 活动和突触传递及其可塑性。这个项目的重点是我们假设的早期 ABO（分钟范围）的影响，触发随后的神经毒性事件，因为我们需要了解所有 Abo毒性。此外，作为早期事件，Abo-b2 AR-S1928信号传导构成了潜在的靶点 用于AD的早期药物干预我们将在一定程度上探讨Cav1.2下游的后续事件。 通过测试6- 48小时范围内的神经毒性是否可以通过Abo-b2 AR-β受体阻断剂改善来检测调节异常。 CaV1.2信号传导和CP-AMPAR在神经元培养物中的作用。我们还将探讨阻断这种信号传导是否会导致 AD 5xFAD小鼠模型将改善这些小鼠中的突触功能。我们肽的开发 破坏b2 AR与Cav1.2的结合是高度创新的，因为它允许特异性阻断信号传导， 通过Abo通过b2 AR而不影响涉及b2 AR的其他信号传导途径。

项目成果

Homeostatic synaptic scaling: molecular regulators of synaptic AMPA-type glutamate receptors.

• DOI: 10.12688/f1000research.13561.1
• 发表时间: 2018
• 期刊: F1000Research
• 作者: Chowdhury D;Hell JW
• 通讯作者: Hell JW

DAPK1 Mediates LTD by Making CaMKII/GluN2B Binding LTP Specific.

• DOI: 10.1016/j.celrep.2017.05.068
• 发表时间: 2017-06-13
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Goodell DJ;Zaegel V;Coultrap SJ;Hell JW;Bayer KU
• 通讯作者: Bayer KU

SynDIG4/Prrt1 Is Required for Excitatory Synapse Development and Plasticity Underlying Cognitive Function.

• DOI: 10.1016/j.celrep.2018.02.026
• 发表时间: 2018-02-27
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Matt L;Kirk LM;Chenaux G;Speca DJ;Puhger KR;Pride MC;Qneibi M;Haham T;Plambeck KE;Stern-Bach Y;Silverman JL;Crawley JN;Hell JW;Díaz E
• 通讯作者: Díaz E

Age-Dependent Contributions of NMDA Receptors and L-Type Calcium Channels to Long-Term Depression in the Piriform Cortex.

NMDA受体和L型钙通道对梨状皮层长期抑郁的年龄依赖性贡献。

• DOI: 10.3390/ijms222413551
• 发表时间: 2021-12-17
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Rajani V;Maziar A;Man KNM;Hell JW;Yuan Q
• 通讯作者: Yuan Q

Loss of SynDIG1 Reduces Excitatory Synapse Maturation But Not Formation In Vivo.

• DOI: 10.1523/eneuro.0130-16.2016
• 发表时间: 2016-09
• 期刊: eNeuro
• 影响因子: 3.4
• 作者: Chenaux G;Matt L;Hill TC;Kaur I;Liu XB;Kirk LM;Speca DJ;McMahon SA;Zito K;Hell JW;Díaz E
• 通讯作者: Díaz E