Pro- and Anti-Nociceptive Actions of P2y Nucleotide Receptors in Sensory Neurons

感觉神经元中 P2y 核苷酸受体的促伤害和抗伤害作用

• 批准号: 7371619
• 负责人: DEREK C MOLLIVER
• 金额: $ 32.48万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7371619
• 关键词: ADP Receptors; ATP Receptors; Acute; Afferent Neurons; Agonist; Analgesics; Animal Model; Behavior; Behavioral; Behavioral Model; Calcium; Clinical Treatment; Cloning; Coupled; Coupling; Development; Esthesia; Failure; Family; Family member; Freund' s Adjuvant; G alpha q Protein; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gated Ion Channel; Genes; Goals; Heating; Hyperalgesia; Image; Immunohistochemistry; Inflammatory; Injection of therapeutic agent; Injury; Investigation; Knockout Mice; Maintenance; Mechanical Stimulation; Mechanics; Mediating; Modeling; Mus; Mutant Strains Mice; Neurons; Nociception; Nociceptors; Normal Statistical Distribution; Nucleotides; Opioid; P2Y2 receptor; Pain; Pain management; Persistent pain; Play; Polymerase Chain Reaction; Reporting; Role; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Spinal Ganglia; Stimulus; Syndrome; TRPV1 gene; Techniques; Testing; Therapeutic Intervention; Thermal Hyperalgesias; allodynia; base; cannabinoid receptor; design; extracellular; genetic manipulation; immunocytochemistry; in vivo; inflammatory pain; insight; member; neurochemistry; purinoceptor P2Y1; purinoceptor P2Y12; receptor; receptor coupling; receptor expression; response

DESCRIPTION (provided by applicant): Extracellular ATP is a messenger in the transduction of noxious stimuli by pain-sensing neurons (nociceptors). The cloning of the sensory neuron-specific ATP-gated ion channel P2X3 generated intense investigation into the ability of ATP to directly activate nociceptors. However, we have new evidence that sensory neurons also express members of a family of G protein-coupled receptors (P2Y receptors) that respond to ATP or related compounds and contribute to nociceptive signaling. These receptors are poorly characterized in neurons: of the 8 known family members only P2Y1 and P2Y2 have been evaluated in sensory neurons and both have been implicated in nociceptive signal transduction. P2Y receptors can be divided into 2 groups based on their coupling to signal transduction pathways; we hypothesize that Gq-coupled receptors are proalgesic while Gi- coupled receptors are analgesic. This proposal consists of 3 Specific Aims designed to identify which P2Y family members contribute to nociceptive signaling and may be useful targets for therapeutic intervention in pain. Specific Aim 1 will use quantitative PCR, immunohistochemistry and Wester blotting techniques to determine which P2Y receptors are expressed in sensory neurons and whether expression changes in response to inflammatory injury. Specific Aim 2 will characterize excitatory and inhibitory actions of Gq-coupled and Gi-coupled P2Y receptors in dissociated sensory neurons using calcium imaging and post-hoc immunocytochemistry. Specific Aim 3 will examine the contribution of P2Y ADP receptors to behavioral pain response thresholds in vivo and will determine whether pharmacological or genetic manipulation of P2Y receptors is analgesic in models of acute and persistent inflammatory pain. These studies will demonstrate the contributions of a new family of receptors to sensory neuron signaling and will provide valuable insight into the mechanisms through which nucleotides act as signaling molecules in the setting of persistent pain.Experiments in this proposal will test the hypothesis that members of the P2Y family of nucleotide receptors are powerful regulators of nociceptor sensitivity that play an important role in the maintenance of persistent pain. We will directly test the possibility that manipulation of these receptors, including the use of antagonists already in development for clinical treatment of non-pain-related syndromes, is an effective analgesic treatment in animal models of persistent pain.

描述（由申请人提供）：细胞外 ATP 是疼痛感知神经元（伤害感受器）转导有害刺激的信使。感觉神经元特异性 ATP 门控离子通道 P2X3 的克隆引发了对 ATP 直接激活伤害感受器能力的深入研究。然而，我们有新的证据表明，感觉神经元也表达 G 蛋白偶联受体（P2Y 受体）家族的成员，这些受体对 ATP 或相关化合物做出反应，并有助于伤害性信号传导。这些受体在神经元中的特征很少：在 8 个已知家族成员中，只有 P2Y1 和 P2Y2 在感觉神经元中进行了评估，并且两者都与伤害性信号转导有关。 P2Y 受体根据其与信号转导途径的耦合可分为 2 组：我们假设 Gq 偶联受体具有促痛作用，而 Gi 偶联受体具有镇痛作用。该提案由 3 个具体目标组成，旨在确定哪些 P2Y 家族成员有助于伤害性信号传导，并可能成为疼痛治疗干预的有用目标。具体目标 1 将使用定量 PCR、免疫组织化学和 Wester blotting 技术来确定哪些 P2Y 受体在感觉神经元中表达，以及表达是否因炎症损伤而发生变化。具体目标 2 将使用钙成像和事后免疫细胞化学来表征分离感觉神经元中 Gq 耦合和 Gi 耦合 P2Y 受体的兴奋和抑制作用。具体目标 3 将检查 P2Y ADP 受体对体内行为疼痛反应阈值的贡献，并将确定 P2Y 受体的药理学或基因操作是否在急性和持续性炎症疼痛模型中具有镇痛作用。这些研究将证明新的受体家族对感觉神经元信号传导的贡献，并将为核苷酸在持续性疼痛中充当信号分子的机制提供有价值的见解。本提案中的实验将检验这样的假设：核苷酸受体的 P2Y 家族成员是伤害感受器敏感性的强大调节剂，在持续性疼痛的维持中发挥着重要作用。我们将直接测试操纵这些受体的可能性，包括使用已经开发的用于非疼痛相关综合征临床治疗的拮抗剂，在持续性疼痛的动物模型中是一种有效的镇痛治疗。