A Female Specific Role of RGSz1 in Modulation of Chronic Pain

RGSz1 在调节慢性疼痛中的女性特异性作用

• 批准号: 9981335
• 负责人: Venetia Zachariou
• 金额: $ 43.26万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9981335
• 关键词: Acute Pain; Affect; Affinity; Attenuated; Behavior; Biochemical; Biochemistry; Biological Assay; Brain; Cell Surface Receptors; Data; Electrophysiology (science); Elements; Enterobacteria phage P1 Cre recombinase; Estrogen Nuclear Receptor; Estrogen Receptors; Female; Fractionation; G-Protein-Coupled Receptors; GTP Phosphohydrolase Activators; GTP-Binding Protein Regulators; GTP-Binding Protein alpha Subunits, Gs; Gene Expression; Gene Silencing; Genes; Genetic; Genetic Transcription; Golgi Apparatus; Gonadal Hormones; Hypersensitivity; Immunofluorescence Immunologic; In Situ; Inflammation; Label; LoxP-flanked allele; Maintenance; Mediating; Messenger RNA; Methodology; Modeling; Molecular; Monitor; Mus; Neurons; Nociception; Norepinephrine; Norepinephrine Receptors; Pain; Pathologic; Pathway interactions; Pattern; Perception; Peripheral; Peripheral nerve injury; Pharmacology; Physiological; Play; Property; Proteins; RGS1 gene; Regulation; Ribosomes; Role; Sensory; Serotonin; Signal Transduction; Spinal Cord Plasticity; Symptoms; Testing; Thermal Hyperalgesias; Transcript; Translating; Viral Vector; Western Blotting; Woman; Work; adeno-associated viral vector; base; central pain; central sensitization; chromatin immunoprecipitation; chronic pain; chronic painful condition; inflammatory neuropathic pain; knock-down; knockout gene; male; mechanical allodynia; men; midbrain central gray substance; mouse model; nerve injury; next generation; novel; overexpression; pain model; pain processing; patch clamp; protein expression; receptor; receptor binding; reduce symptoms; response; serotonin receptor; sex; transcriptome sequencing; transmission process; vector

SUMMARY While the mechanisms modulating the transmission and perception of chronic pain differ between men and women, the exact neuroanatomical and molecular differences existing between the sexes remain only partly understood. This proposal focuses on the study of intracellular mechanisms that modulate symptoms of long-term pain states in female mice. We have identified signal transduction mechanisms in the mouse ventrolateral periaqueductal gray (vlPAG) that selectively modulate sensory symptoms of inflammatory and neuropathic pain in female mice. The signal transduction modulator Regulator of G Protein Signaling-1 (RGSz1) controls the function of G protein coupled receptors (GPCRs) by binding to activated Gi subunits, including the Golgi enriched Gz. RGSz1 negatively modulates the amplitude and direction of signal transduction of several GPCRs involved in the modulation of pain processing, including the serotonin 5HT1A receptor. We recently found that RGSz1 mRNA and protein levels are dynamically regulated in the vlPAG of female (but not male) mice by long-term peripheral inflammation. Constitutive deletion of the RGSz1 gene or vlPAG-specific knockdown of RGSz1, exacerbate sensory hypersensitivity behaviors such as thermal hyperalgesia and mechanical allodynia in female mice, but they have no effect in male mice. We will investigate the mechanisms regulating the expression of RGSz1 in the male and female vlPAG at various points after the induction of peripheral nerve injury or inflammation. We will apply several genetic mouse models for regional inactivation or overexpression of RGSz1 in vlPAG neuronal subsets, along with brain biochemistry, electrophysiology and voltammetry to understand the sex-specific role of RGSz1 in the function of the descending inhibitory pathway in models of chronic pain. Since RGSz1 plays a prominent role in the function of the Golgi apparatus, will apply chronic pain models to understand the impact of RGSz1 on the expression levels of trans- Golgi components in the presence and in the absence of pain. Finally, we will use RNA Sequencing to understand the impact of RGSz1 on gene expression adaptations underlying chronic pain states.

概括 虽然调节慢性疼痛的传递和感知的机制不同 男性和女性之间存在确切的神经解剖学和分子差异 两性之间的关系仍然只有部分了解。本提案的重点是研究 调节雌性小鼠长期疼痛状态症状的细胞内机制。我们 确定了小鼠腹外侧导水管周围灰质的信号转导机制 (vlPAG) 选择性调节炎症和神经性疼痛的感觉症状 雌性小鼠。信号转导调节剂 G 蛋白信号传导调节剂 - 1 (RGSz1) 通过与激活的 Gi 结合来控制 G 蛋白偶联受体 (GPCR) 的功能 亚基，包括高尔基体富集的 Gz。 RGSz1 负向调制幅度并 参与疼痛处理调节的几个 GPCR 的信号转导方向， 包括血清素 5HT1A 受体。我们最近发现RGSz1 mRNA和蛋白 雌性（而非雄性）小鼠的 vlPAG 水平通过长期动态调节 周围炎症。 RGSz1 基因的组成型缺失或 vlPAG 特异性敲低 RGSz1，加剧感觉超敏行为，例如热痛觉过敏和 对雌性小鼠有机械性异常性疼痛，但对雄性小鼠没有影响。我们将调查 不同条件下雄性和雌性 vlPAG 中 RGSz1 表达的调节机制 诱发周围神经损伤或炎症后的穴位。我们会申请几个 vlPAG 神经元中 RGSz1 区域失活或过度表达的遗传小鼠模型 子集，以及大脑生物化学、电生理学和伏安法，以了解 RGSz1 在下行抑制途径功能中的性别特异性作用 慢性疼痛。由于 RGSz1 在高尔基体的功能中起着突出的作用，因此 应用慢性疼痛模型来了解 RGSz1 对反式表达水平的影响 存在和不存在疼痛时的高尔基体成分。最后，我们将使用RNA 测序以了解 RGSz1 对基因表达适应的影响 慢性疼痛状态。