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.

抽象的 β 淀粉样肽导致 Cav1.2 失调 L 型 Ca2+ 通道 (LTCC) 是基因转录、神经元兴奋性和突触的关键调节因子 Cav1.2 是大脑中普遍存在的 LTCC（Hell et al., 1993, JCB 123, 949-962）。 Ca2+ 通过 LTCC 流入很早就与老年症状和阿尔茨海默病 (AD) 有关（例如， Science 272, 1017）我们发现老年大鼠的 PKA 介导的磷酸化显着增加。 丝氨酸 1928 上的 Cav1.2 (Davare and Hell, 2003, PNAS 100, 16018-23)，增加 Cav1.2 通道活性 (Qian, …, Hell, 2017, Sci Sig 10, eaaf9659) 我们还发现 Cav1.2 与 形成独特的信号复合体。 b2 肾上腺素受体 (b2 AR) 还包含 cAMP 介导的所有其他必需蛋白质 Cav1.2 的调节（例如，Davare 等人，2001，Science 293, 98），使 Cav1.2 成为 b2 AR 信号传导的主要目标。 b 淀粉样肽 1-42 (Ab) 是 AD 的一个标志，寡聚 Abo 会刺激 b2 AR。 单通道记录表明 Abo 通过 b2 AR 增加 Cav1.2 活性。 增加是由 Cav1.2 的主要 PKA 位点 S1928 磷酸化介导的，并触发下游事件 最终导致神经元损伤的目标 1 是测试 Abo 是否通过 Cav1.2- 调节 Cav1.2。 相关的 b2 AR 发生在树突和棘中（Ca2+ 成像），无论其部分是由增加的介导的 Cav1.2 的表面插入，以及是否通过药理学或创新的 S1928A 阻断该信号传导 敲入 (KI) 小鼠减轻 Abo 神经毒性 目的 2 是测试 Abo - b2 AR - S1928 信号传导是否受到刺激。 Ca2+ 可渗透 (CP) AMPAR（GluA1 同聚物）的表面插入和阻断 CP-AMPAR 可减轻 Abo 我们将使用 SEP 标记的 GluA1 和 GluA2 的尖端实时成像并分析现场 EPSP。 和 mEPSC 在谷氨酸再摄取抑制之前和之后测量突触周围的 AMPAR 活性 目标 3 将测试 Abo - b2 AR - S1928 信号在 Abo 增强长期抑郁症中的作用， 和长期增强作用，Abo 与我们的 S1928A KI 小鼠杂交会损害这种增强作用。 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 这个项目的重点是我们早期从事的。 Abo（分钟范围）的效应会触发随后的神经毒性事件，因为我们需要了解所有 Abo 毒性方面此外，作为早期事件，Abo - b2 AR - S1928 信号传导构成了潜在的目标。 对于 AD 的早期药物干预，我们将在某种程度上探索 Cav1.2 的后期下游事件。 通过测试 Abo - b2AR - 阻滞剂是否可以改善 6-48 小时范围内的神经毒性来调节失调 我们还将探讨是否在神经培养物中阻断该信号传导。 AD 的 5xFAD 小鼠模型将改善这些小鼠的突触功能 我们的肽的开发。 破坏 b2 AR 与 Cav1.2 结合的方法具有高度创新性，因为它可以特异性阻断信号传导 Abo 通过 b2 AR 产生信号，而不影响涉及 b2 AR 的其他信号通路。

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

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

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

• 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