the The Involvement of Respiration-Related Neurones in the Medullary Respiratory Rhythm Generation Center in the Opioid-Induced Respiratory Depression

延髓呼吸节律发生中心的呼吸相关神经元参与阿片类药物引起的呼吸抑制

• 批准号: 13671557
• 负责人: TAKITA Koichi
• 金额: $ 2.11万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671557/
• 关键词: Respirtory neurones; Whole cell recording; Blind patch clamp; New born rat; Immunohistochemistry; Opioid; Nociceptin; orphanin FQ; ブラインドパッチクランプ法; Mu-オピオイド受容体; ノシセプチン; 免疫組織染色; C-Fos; 中枢性化学受容体

In brainstem-spinal cord preparations isolated from newborn rats, whole cell recording technique by blind patch clamp method was applied to record membrane potentials of respiration-related neurones in the medullary respiratory rhythm generating center. Neurones recorded was marked with 0.5% neurobiotin. To examine whether the neurones marked with neurbiotin have μ-opioid receptors, immunohistochemistry of μ-opioid receptors was also performed. In the present study, we have established the whole cell recording technique in brainstem-spinal cord preparations isolated from newborn rats, and the whole cell recordings from Type I, Type II, Type III inspiratory neurones and pre-inspiratory neurones in the medullary respiration center were successfully achieved. However, these respiratroy related-neurones did not show immunoreactivity of μ-opioid receptors.To elucidate the roles nociceptin/orphanin FQ (N/OFQ) and the nociceptin/orphanin FQ peptide (NOP) receptor (previously termed ORL1 receptor) play in the medullary structures in the generation of respiratory rhythm, the respiratory effects of N/OFQ and the NOP antagonist, Comp B were examined, using in vitro newborn rat preparations. N/OFQ reduced fR in a concentration-dependent manner. Pretreatment with CompB 10μM prevented the N/OFQ 10 nM-induced fR reduction, whereas CompB itself did not affect fR. These results show that N/OFQ acts as a neuromodulator to reduce fR in the respiratory rhythm-generating center in the medulla oblongata, and this action of N/OFQ is mediated by NOP receptors.

在从新生大鼠分离的脑干脊髓标本中，采用盲膜片钳法的全细胞记录技术来记录髓质呼吸节律发生中心的呼吸相关神经元的膜电位，并用0.5％神经生物素标记记录的神经元。神经生物素标记的神经元是否具有μ阿片受体，本研究还进行了μ阿片受体的免疫组织化学分析，建立了全细胞记录技术。从新生大鼠中分离出脑干脊髓制剂，并成功获得了延髓呼吸中枢的I型、II型、III型吸气神经元和吸气前神经元的全细胞记录，但这些呼吸相关神经元没有显示。 μ-阿片受体的免疫反应性。阐明伤害肽/孤啡肽 FQ (N/OFQ) 和伤害肽/孤啡肽的作用FQ 肽 (NOP) 受体（以前称为 ORL1 受体）在呼吸节律产生中的髓质结构中发挥作用，使用体外新生大鼠制剂检查了 N/OFQ 和 NOP 拮抗剂 Comp B 的呼吸作用。 OFQ 以浓度依赖性方式降低 fR，用 CompB 10μM 预处理可防止 N/OFQ 10 nM 诱导的 fR 降低，而 CompB 本身不影响 fR。研究表明，N/OFQ 作为神经调节剂，降低延髓呼吸节律发生中心的 fR，并且 N/OFQ 的这种作用是由 NOP 受体介导的。

项目成果

Takita K, Morimoto Y, Kemmotsu O.: "The roles of nociceptin/orphanin FQ and nociceptin/orphanin FQ peptide (NOP) receptor in respiratory rhythm generation in the medulla oblongata : an in vitro study"British Journal of Anaesthesia. (in press).

Takita K、Morimoto Y、Kemmotsu O.：“伤害感受肽/孤啡肽 FQ 和伤害感受肽/孤啡肽 FQ 肽 (NOP) 受体在延髓呼吸节律生成中的作用：体外研究”英国麻醉杂志。

Takita K., Morimoto Y., Kemmotsu O.: "Roles of nociceptin/orphanin FQ and nociceptin/orphanin FQ peptide receptor in respiratory rhythm generation in the medulla oblongata: an in vitro study"British Journal of Anaesthesia. (in press).

Takita K.、Morimoto Y.、Kemmotsu O.：“伤害感受肽/孤啡肽 FQ 和伤害感受肽/孤啡肽 FQ 肽受体在延髓呼吸节律生成中的作用：一项体外研究”英国麻醉杂志。