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

基本信息

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
项目类别：
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&apos 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

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&apos 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

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项目摘要

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是通过疼痛感应神经元（伤害感受器）转导有害刺激的使者。感觉神经元特异性ATP门控离子通道P2X3的克隆产生了对ATP直接激活伤害感受器的能力的强烈研究。但是，我们有新的证据表明，感觉神经元还表达了对ATP或相关化合物反应的G蛋白偶联受体（P2Y受体）家族的成员，并有助于伤害感受信号传导。这些受体在神经元中的特征很差：在8个已知家庭成员中，仅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受体可以根据信号转导途径的耦合将其分为两组。我们假设GQ耦合受体是促痛的，而Gi-C偶联受体是镇痛的。该提案包括3个特定目标，旨在确定哪种P2Y家庭成员有助于伤害性信号，并且可能是治疗干预疼痛的有用靶标。具体目标1将使用定量PCR，免疫组织化学和Wester印迹技术来确定哪些P2Y受体在感觉神经元中表达了哪些P2Y受体，以及是否对炎症性损伤的反应而表达是否变化。特定的目标2将使用钙成像和事后免疫细胞化学化学的GQ耦合和GI耦合P2Y受体的GQ耦合和GI耦合P2Y受体的兴奋性和抑制作用。特定的目标3将检查P2Y ADP受体对体内行为疼痛反应阈值的贡献，并确定P2Y受体的药理或遗传操纵是否在急性和持续性炎症性疼痛的模型中是镇痛作用。这些研究将证明新的受体家族对感觉神经元信号的贡献，并将为核苷酸在持续性疼痛的环境中充当信号分子的机制提供宝贵的见解。该提案中的经验将检验以一种强大的调节剂的态度，对核苷酸的p2y家族的作用有强大的作用，对nocicepteptors的作用有强大的作用。我们将直接测试对这些受体操纵的可能性，包括在临床治疗非伴侣相关综合症的临床治疗中使用拮抗剂是一种有效的镇痛治疗。