【作者】 王宁黔；

【导师】 吴中海；

【作者基本信息】 第一军医大学 ， 神经生理学， 2002， 硕士

【摘要】 目的 1)应用HRP／WGA-HRP逆行/顺行束路追踪技术，对面神经后核内侧区(the medial area of retro facialis nucleus，mNRF)与脑干呼吸相关区域的纤维投射联系进行研究，探讨基本呼吸节律发生及调控的解剖学基础；2)应用新生大鼠离体延髓脑片标本，同步记录舌下神经根和mNRF的呼吸神经元单位的放电活动，进一步明确mNRF在基本呼吸节律发生中的作用：观察甘氨酸(Glycine，Gly)类受体相应的激动剂和拮抗剂对吸气神经元放电活动的影响，进一步探讨其在基本呼吸节律发生和调控中的作用。 方法 1)选用成年SD大鼠，体重200-250g，雌雄不拘。参照图谱，将HRP／WGA-HRP微量注射入mNRF，动物存活48小时后，灌注固定，取脑，冰冻切片，DAB／TMB方法成色，参照图谱光镜观察阳性胞体或纤维的所在区域。2)选用新生SD大鼠(0-3d)，雌雄不拘。参照并改良Suzue的方法制作离体延髓脑片标本，记录舌下神经放电作为呼吸活动的指标，并在延髓腹外侧mNRF区同步记录呼吸神经元单位的放电活动。并通过灌流液给药，观察不同药物对吸气神经元放电活动的影响。 结果 1)发现HRP标记神经元胞体主要位于双侧延髓孤束核(NTS)、疑核(AMB)、巨细胞网状核(Gi)、旁巨细胞网状背核(DpGi)、中间网状核(IRt)、外侧网状核(LRt)、中缝核群(raphe muclei)、桥脑臂旁核(PB)、K-F核、脊髓中间区(spinal intermediate zone)、脊髓后角、延髓及桥脑网状结构；2)WGA-HRP标记神经纤维主要存在于双侧孤束核、疑核、脊髓中间区、延髓网状结构；3)在面神经后核内侧区内可记录到多种类型的呼吸神经元放电活动；4)正常对照MKS液灌流时，mNRF区吸气神经元放电的峰频率为28.80±2.77次／s，以Glycine受体抑制剂士的宁(STR)灌流5分钟后，峰频率上升到38.60±4.33次／s 0＜0．01X后以Glycine受体激动剂甘氨酸（Glycine，Gly）灌流5分钟 后，峰频率下降至251513．42次儿o＜0刀5人 最后以MKS液洗脱，峰 频率回升到 31．7616．80次／s。 结论 以上1L2）结果提示，在成年SD大鼠标本上：mNRF与脑 干不同呼吸相关区域之间具有广泛的纤维投射联系。在基本呼吸节律发 生及维持过程中，脑干呼吸相关区域与mNRF可能通过这些神经结构的 相互作用对基本呼吸节律进行调控。 结果3）提示，在新生SD大鼠延髓脑片标本上mw区内存在各 类型的呼吸神经元。 通过结果4）可以看出：激活或抑制甘氨酸受体可以明显改变mN’RF 区吸气神经元的放电活动，调节放电的峰频率。提示：甘氨酸受体介导 了面神经后核内侧区吸气神经元的抑制性突触输入，并可能在基本呼吸 节律的发生中发挥重要抑制作用。更多还原

【Abstract】 Objectl.To illustrate the afferent/efferent projections of cells in the medial area of nucleus retrofacialis (mNRF) in the adult SD rat using retrograde/ anterograde tracing methods;2. To investigate the roles of glycine receptors in the primary respiratory rhythm generation and modulation by exploring the effects of glycine receptors agonist and antagonist on the discharge activities of inspiratory neurons in mNRF. Methods:1. Twenty-five adult SD rats of either sex,weighting 200-250g,were used in the first part of study. According to the atlas,HRP/WGA-HRP solution was microinjected with a microsyringe into mNRF unilaterally. Control:HRP solution was microinjected into the area 2mm away from mNRF. After a survival time of 48h,animals were perfused transcardially to fix the brain. The brains were removed and then frozen,sliced into 40(xm transverse sections. The sections were processed histochemically according to the tetramethylbenzidine(TMB) method. After reactions,the sections were mounted on gelatin-coated slides. HRP/WGA-HRP products were examined microscopically under bright-field illumination.2. The second part studies were performed on the in vitro brainstem slice isolated from neonatal rats (0-3 d). The respiratory activity was monitored by suction electrode applied on the central end of hypoglossal nerve and the respiratory neuronal discharges were recorded extracellularly by inserting theglass microelectrode into the mNRF of the slices. The inspiratory neurons were determined according to the temporal relationship between the neuronal discharge and the hypoglossal nerve activity. Drugs were administered by bath application and their effects on the inspiratory neuronal activity were investigated. Results:l.HRP labeled cell bodies were mainly found in AMB,NTS,Gi,raphe nuclei,PB-KF complex,spinal intermediate zone and reticular formation of medulla oblongata and pons.2.WGA-HRP labeled terminal fibers were mainly observed in AMB,NTS,intermediate zone and reticular formation of pons.S.Inspiratory/expiratory neuronal discharge activities were recorded in mNRF extracellularly.4. During the control perfusion,the peak frequency of inspiratory neurons was 28.80+2.77Hz. After application of STR,the peak frequency was increased to 38.60+4.33Hz (P<0.01). After application of Glycine,the peak frequency was decreased to 25.15+3.42Hz (P<0.05). After washout,the peak frequency was 31.76+6.80Hz. Conclusions:1 The first 2 results indicated that in the adult rats:1) There are widely anatomical connections between many respiratory related regions in brainstem and mNRF;2) These connections between many respiratory related regions in brainstem and mNRF may be involved in the adjustment of the basic respiratory rhythm generation.The other 2 results indicated that in the in vitro brainstem slice from neonatal rats:1) All subtypes of respiratory neurons located in mNEF. 2) Activation/inhibition of Glycine receptors can affect the inspiratory neuronal discharge in mNRF,which indicated that Glycine is involved in the generation and maintenance of the basic respiratory rhythm as an inhibitor.更多还原