Molecular mechanisms of store-operated calcium channels in pain

钙池操纵的钙通道在疼痛中的分子机制

• 批准号: 9317548
• 负责人: Huijuan Hu
• 金额: $ 34.23万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9317548
• 关键词: Adult; Agonist; Analgesics; Attenuated; Behavior; Brain; Ca(2+)-Transporting ATPase; Calcium; Calcium Channel; Cell membrane; Cell physiology; Cells; Central Nervous System Diseases; Data; Development; Endoplasmic Reticulum; Formalin; Freund' s Adjuvant; Goals; Grant; Hypersensitivity; Image; Individual; Intervention; Ionomycin; Knock-out; Knowledge; Link; Mediating; Methods; Microscope; Modeling; Molecular; Mus; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; Neuraxis; Neuroglia; Neurons; Neuropathy; Nociception; Pain; Pathologic; Pharmacology; Play; Posterior Horn Cells; Property; Proteins; Public Health; Research; Role; STIM1 gene; Signal Transduction; Slice; Small Interfering RNA; Synapses; Synaptic Transmission; Synaptic plasticity; Testing; Thapsigargin; Therapeutic; Therapeutic Intervention; Time; Western Blotting; base; behavior test; central sensitization; chronic pain; inflammatory pain; inhibitor/antagonist; insight; interest; knock-down; live cell imaging; nerve injury; neuronal excitability; neurotransmitter release; novel; novel strategies; painful neuropathy; patch clamp; public health relevance; response; sensor; spared nerve; tool

DESCRIPTION (provided by applicant): Chronic pain continues to be a major public health concern with considerable effort directed towards achieving a better understanding of the underlying mechanisms to facilitate the development of more effective and safer therapeutics. The research in my lab focuses on the role of store-operated calcium channels (SOCCs) in the modulation of pain. SOC channels are composed of recently discovered Orai subunits (Orai1/2/3, pore-forming subunits), and stromal interaction molecules (STIM) 1 and STIM2 (the ER Ca2+ sensors, serve as their activators). Activation of SOCCs by the release of calcium from endoplasmic reticulum (ER) leads to sustained high levels of cytosolic Ca2+ that are required for many calcium-dependent cellular processes. In the central nervous system (CNS), SOC channels are known to influence neurotransmitter release and synaptic plasticity. Results from our current R21 grant have demonstrated that SOCCs are functionally expressed in dorsal horn neurons and their pharmacological properties are similar to those in non-excitable cells. We have identified STIM1 and Orai1 as essential components of SOCCs in dorsal horn neurons and have demonstrated that the knockdown of STIM1 and Orai1 proteins individually reduces spare nerve injury (SNI)-induced neuropathic pain. Recently, we found that the SOC inhibitor YM- 58483 not only attenuates neuropathic pain, but also dramatically diminishes inflammatory pain. Of interest is the finding that a SOCC inhibitor blocks NMDA and mGluR1/5 agonist- induced sustained calcium entry and nociceptive behavior at later time points, suggesting that SOCCs are involved in functions of NMDA receptors and mGluR1/5. Based on these findings, we hypothesize that SOCC involvement in pain is mediated by their interaction with NMDA receptors and mGluR1/5. To test this hypothesis, we will combine the methods of patch-clamp recording, calcium imaging recording, Western Blot analysis and behavioral testing. Overall, our long-term goal is to understand the mechanisms of chronic pain. The goals of this proposal are to identify endogenous upstream molecules of SOCC signaling and explore functional consequences of SOCC activation. These studies will extend our knowledge of how the SOCC signaling modulates pain. Our findings will establish functional links between SOCCs and group I mGluR, and between SOCCs and NMDA receptors. These studies will provide novel insights into mechanisms of pain and other CNS disorders and may suggest new approaches to therapeutic intervention.

描述（由申请人提供）：慢性疼痛仍然是一个主要的公共卫生问题，相当大的努力旨在更好地了解潜在的机制，以促进更有效和更安全的治疗方法的发展。我实验室的研究重点是钙库操纵的钙通道（SOCCs）在疼痛调节中的作用。SOC通道由最近发现的奥赖亚基（Orai 1/2/3，孔形成亚基）和基质相互作用分子（STIM）1和STIM 2（ER Ca 2+传感器，充当它们的激活剂）组成。通过从内质网（ER）释放钙激活SOCC导致持续高水平的胞质Ca 2+，这是许多钙依赖性细胞过程所需的。在中枢神经系统（CNS）中，已知SOC通道影响神经递质释放和突触可塑性。从我们目前的R21资助的结果表明，SOCCs在背角神经元中功能性表达，其药理学性质与非兴奋细胞中的相似。我们已经确定了STIM 1和Orai 1作为背角神经元中SOCC的重要组成部分，并证明了STIM 1和Orai 1蛋白的敲除分别减少了备用神经损伤（SNI）诱导的神经性疼痛。最近，我们发现SOC抑制剂YM- 58483不仅能减轻神经性疼痛，而且能显著减轻炎症性疼痛。令人感兴趣的是发现SOCC抑制剂在稍后的时间点阻断NMDA和mGluR 1/5激动剂诱导的持续钙内流和伤害感受行为，表明SOCC参与NMDA受体和mGluR 1/5的功能。基于这些发现，我们假设SOCC参与疼痛是通过它们与NMDA受体和mGluR 1/5的相互作用介导的。为了验证这一假设，我们将结合联合收割机的方法，膜片钳记录，钙成像记录，Western Blot分析和行为测试。总的来说，我们的长期目标是了解慢性疼痛的机制。该提案的目标是确定SOCC信号传导的内源性上游分子，并探索SOCC激活的功能后果。这些研究将扩展我们对SOCC信号如何调节疼痛的认识。我们的研究结果将建立SOCCs和I组mGluR之间的功能联系，以及SOCCs和NMDA受体之间的功能联系。这些研究将为疼痛和其他中枢神经系统疾病的机制提供新的见解，并可能提出新的治疗干预方法。