Objective Aims to delineate the distribution profile of three isoforms of vesicular glutamate transporter （VGluT）, viz. VGluT1-3, and their cellular localization within vestibular nuclear complex （VNC）. Methods Br...Objective Aims to delineate the distribution profile of three isoforms of vesicular glutamate transporter （VGluT）, viz. VGluT1-3, and their cellular localization within vestibular nuclear complex （VNC）. Methods Brain sections from normal Sprague-Dawley rats were processed immunohistochemically for VGluT detection, employing avidinbiotinylated peroxidase complex method with 3-3＇-diaminobenzidine （DAB） as chromogen. Results The whole VNC expressed all of the three transporters that were observed to be localized to the fiber endings. Compared with VGluT1 and VGluT3, VGluT2 demonstrated a relatively homogeneous distribution, with much higher density in VNC. VGluT3 displayed the highest density in lateral vestibular nucleus and group X, contrasting with the sparse immunostained puncta within vestibular medial and inferior nuclei. Conclusion Glutamtatergic pathways participate in the processing of vestibular signals within VNC mainly through the re-uptake of glutamate into synaptic vesicles by VGluT1 and 2, whereas VGluT3 may play a similar role mainly in areas other than medial and inferior nuclei of VNC.展开更多
目的观察慢性束缚应激后,小鼠清醒状态下咬肌肌电水平以及支配咬肌运动的三叉神经运动核(trigeminal motor nucleus,Vmo)神经元的变化,为探究心理因素与颞下颌关节紊乱病发生的相关中枢调控机制提供实验依据。方法32只雄性小鼠被随机分...目的观察慢性束缚应激后,小鼠清醒状态下咬肌肌电水平以及支配咬肌运动的三叉神经运动核(trigeminal motor nucleus,Vmo)神经元的变化,为探究心理因素与颞下颌关节紊乱病发生的相关中枢调控机制提供实验依据。方法32只雄性小鼠被随机分为对照组、应激组,对应激组小鼠施加4 h/d、连续14 d的慢性束缚应激;对照组小鼠正常饲养。14 d后,通过旷场实验与高架十字迷宫实验观察小鼠的行为学改变;检测清醒状态下小鼠咬肌肌电水平;采用全细胞膜片钳技术观察Vmo神经元的电生理特性,并利用免疫组织荧光技术观察Vmo内Ⅰ型、Ⅱ型囊泡膜谷氨酸转运体(vesicular glutamate transporter 1/2,VGLUT1/2)的表达情况。结果应激组小鼠在旷场实验的中央活动时间(P=0.0004)与中央活动路程(P=0.0004)均显著低于对照组;高架十字迷宫实验中应激组小鼠的开臂进入次数百分比(P=0.0002)与滞留开臂时间百分比(P=0.0013)均显著低于对照组,显示存在明显的焦虑样行为。对照组和应激组小鼠在应激开始前,咬肌累积肌电(integral electromyography,iEMG)(P=0.8779)及振幅均方根(root mean square,RMS)(P>0.9999)均无明显差异;应激结束后,应激组小鼠咬肌的iEMG(P=0.0004)和RMS值(P=0.0001)均显著高于对照组。对照组小鼠在应激前后的iEMG(P=0.7989)和RMS值(P>0.9999)比较无显著差异;应激组小鼠在应激结束后,其咬肌的iEMG(P=0.0011)和RMS值(P=0.0019)显著高于应激前水平。电生理结果显示,在电流钳模式下,当输入60、80、100 pA电流时,应激组小鼠Vmo神经元的放电频率显著高于对照组(P<0.05);应激组小鼠Vmo神经元的自发性兴奋性突触后电流频率(P=0.0030)与幅度(P=0.0002)显著高于对照组。免疫组织荧光染色结果显示,应激组小鼠Vmo部位的VGLUT1(P=0.0010)与VGLUT2荧光强度(P=0.0013)均显著高于对照组。结论慢性束缚应激能够导致小鼠的焦虑样行为及咬肌肌电活动�展开更多
Activation of inflammatory responses regulates the transmission of pain pathways through an integrated network in the peripheral and central nervous systems.The immunopotentiator thymosin alpha-1(Tal)has recently been...Activation of inflammatory responses regulates the transmission of pain pathways through an integrated network in the peripheral and central nervous systems.The immunopotentiator thymosin alpha-1(Tal)has recently been reported to have anti-inflammatory and neuroprotective functions in rodents.However,how Tα1 affects inflammatory pain remains unclear.In the present study,intraperitoneal injection of Tal attenuated complete Freund's adjuvant(CFA)-induced pain hypersensitivity,and decreased the up-regulation of pro-inflammatory cytokines(TNF-α,IL-1β,and IL-6)in inflamed skin and the spinal cord.We found that CFA-induced peripheral inflammation evoked strong microglial activation,but the effect was reversed by Tα1.Notably,Tα1 reversed the CFA-induced up-regulation of vesicular glutamate transporter(VGLUT)and down-regulated the vesicular γ-aminobutyric acid transporter(VGAT)in the spinal cord.Taken together,these results suggest that Tα1 plays a therapeutic role in inflammatory pain and in the modulation of microgliainduced pro-inflammatory cytokine production in addition to mediation of VGLUT and VGAT expression in the spinal cord.展开更多
Previous studies have shown that electroacupuncture（EA） promotes recovery of motor function in Parkinson＇s disease（PD）. However the mechanisms are not completely understood. Clinically, the subthalamic nucleus（...Previous studies have shown that electroacupuncture（EA） promotes recovery of motor function in Parkinson＇s disease（PD）. However the mechanisms are not completely understood. Clinically, the subthalamic nucleus（STN） is a critical target for deep brain stimulation treatment of PD, and vesicular glutamate transporter 1（VGlu T1） plays an important role in the modulation of glutamate in the STN derived from the cortex. In this study,a 6-hydroxydopamine（6-OHDA）-lesioned rat model of PD was treated with 100 Hz EA for 4 weeks. Immunohistochemical analysis of tyrosine hydroxylase（TH） showed that EA treatment had no effect on TH expression in the ipsilateral striatum or substantia nigra pars compacta,though it alleviated several of the parkinsonian motor symptoms. Compared with the hemi-parkinsonian rats without EA treatment, the 100 Hz EA treatment significantly decreased apomorphine-induced rotation and increased the latency in the Rotarod test. Notably, the EA treatment reversed the 6-OHDA-induced down-regulation of VGlu T1 in the STN. The results demonstrated that EA alleviated motor symptoms and up-regulated VGlu T1 in the ipsilateral STN of hemi-parkinsonian rats, suggesting that up-regulation of VGlu T1 in the STN may be related to the effects of EA on parkinsonian motor symptoms via restoration of function in the cortico-STN pathway.展开更多
AIM:To investigate the activity of vesicular glutamate transporter-3(VGLUT3) in a visceral hyperalgesia rat model of irritable bowel syndrome,and the role of mast cells(MCs).METHODS:Transient intestinal infection was ...AIM:To investigate the activity of vesicular glutamate transporter-3(VGLUT3) in a visceral hyperalgesia rat model of irritable bowel syndrome,and the role of mast cells(MCs).METHODS:Transient intestinal infection was inducedby oral administration of Trichinella spiralis larvae in rats.On the 100th day post-infection(PI),the rats were divided into an acute cold restraint stress(ACRS)group and a non-stressed group.Age-matched untreated rats served as controls.The abdominal withdrawal reflex was used to measure the visceromotor response to colorectal distension(CRD).The expression levels of VGLUT3 in peripheral and central neurons were analyzed by immunofluorescence and western blotting.RESULTS:VGLUT3 expression in the L6S1 dorsal root ganglion cells was significantly higher in the PI group than in the control group(0.32±0.009 vs0.22±0.008,P<0.01),and there was no significant difference in the expression of VGLUT3 between MCdeficient rats and their normal wild-type littermates.Immunofluorescence showed that the expression levels of VGLUT3 in PI+ACRS rats were enhanced in the prefrontal cortex of the brain compared with the control group.CONCLUSION:VGLUT3 is involved in the pathogenesis of visceral hyperalgesia.Coexpression of c-fos,5-hydroxytryptamine and VGLUT3 after CRD was observed in associated neuronal pathways.Increased VGLUT3 induced by transient intestinal infection was found in peripheral nerves,and was independent of MCs.Moreover,the expression of VGLUT3 was enhanced in the prefrontal cortex in rats with induced infection and stress.展开更多
基金National Natural Science Foundation of China （No. 30470562）.
文摘Objective Aims to delineate the distribution profile of three isoforms of vesicular glutamate transporter （VGluT）, viz. VGluT1-3, and their cellular localization within vestibular nuclear complex （VNC）. Methods Brain sections from normal Sprague-Dawley rats were processed immunohistochemically for VGluT detection, employing avidinbiotinylated peroxidase complex method with 3-3＇-diaminobenzidine （DAB） as chromogen. Results The whole VNC expressed all of the three transporters that were observed to be localized to the fiber endings. Compared with VGluT1 and VGluT3, VGluT2 demonstrated a relatively homogeneous distribution, with much higher density in VNC. VGluT3 displayed the highest density in lateral vestibular nucleus and group X, contrasting with the sparse immunostained puncta within vestibular medial and inferior nuclei. Conclusion Glutamtatergic pathways participate in the processing of vestibular signals within VNC mainly through the re-uptake of glutamate into synaptic vesicles by VGluT1 and 2, whereas VGluT3 may play a similar role mainly in areas other than medial and inferior nuclei of VNC.
基金supported by the Foundation for Distinguished Young Talents in Higher Education of Guangdong Province, China (2016KQNCX019 and 2016KQNCX027)the National Natural Science Foundation of China (31571041)+1 种基金the Guangdong Provincial Department of Education Innovating Strong National Engineering Major Project (2014GKXM031)Guangdong Provincial Universities and Colleges Pearl River Scholar Funded Scheme (2016)
文摘Activation of inflammatory responses regulates the transmission of pain pathways through an integrated network in the peripheral and central nervous systems.The immunopotentiator thymosin alpha-1(Tal)has recently been reported to have anti-inflammatory and neuroprotective functions in rodents.However,how Tα1 affects inflammatory pain remains unclear.In the present study,intraperitoneal injection of Tal attenuated complete Freund's adjuvant(CFA)-induced pain hypersensitivity,and decreased the up-regulation of pro-inflammatory cytokines(TNF-α,IL-1β,and IL-6)in inflamed skin and the spinal cord.We found that CFA-induced peripheral inflammation evoked strong microglial activation,but the effect was reversed by Tα1.Notably,Tα1 reversed the CFA-induced up-regulation of vesicular glutamate transporter(VGLUT)and down-regulated the vesicular γ-aminobutyric acid transporter(VGAT)in the spinal cord.Taken together,these results suggest that Tα1 plays a therapeutic role in inflammatory pain and in the modulation of microgliainduced pro-inflammatory cytokine production in addition to mediation of VGLUT and VGAT expression in the spinal cord.
基金supported by the Beijing Municipal Science and Technology Commission (Z161100002616007)National Key Research and Development Program (2016YFC1306300)+1 种基金the Major Program of the National Natural Science Foundation of China (81527901)the Natural Science Foundation of Beijing Municipality (7082008)
文摘Previous studies have shown that electroacupuncture（EA） promotes recovery of motor function in Parkinson＇s disease（PD）. However the mechanisms are not completely understood. Clinically, the subthalamic nucleus（STN） is a critical target for deep brain stimulation treatment of PD, and vesicular glutamate transporter 1（VGlu T1） plays an important role in the modulation of glutamate in the STN derived from the cortex. In this study,a 6-hydroxydopamine（6-OHDA）-lesioned rat model of PD was treated with 100 Hz EA for 4 weeks. Immunohistochemical analysis of tyrosine hydroxylase（TH） showed that EA treatment had no effect on TH expression in the ipsilateral striatum or substantia nigra pars compacta,though it alleviated several of the parkinsonian motor symptoms. Compared with the hemi-parkinsonian rats without EA treatment, the 100 Hz EA treatment significantly decreased apomorphine-induced rotation and increased the latency in the Rotarod test. Notably, the EA treatment reversed the 6-OHDA-induced down-regulation of VGlu T1 in the STN. The results demonstrated that EA alleviated motor symptoms and up-regulated VGlu T1 in the ipsilateral STN of hemi-parkinsonian rats, suggesting that up-regulation of VGlu T1 in the STN may be related to the effects of EA on parkinsonian motor symptoms via restoration of function in the cortico-STN pathway.
基金Supported by National Natural Science Foundation of China,No.30940033the Doctoral Program of Higher Education of China
文摘AIM:To investigate the activity of vesicular glutamate transporter-3(VGLUT3) in a visceral hyperalgesia rat model of irritable bowel syndrome,and the role of mast cells(MCs).METHODS:Transient intestinal infection was inducedby oral administration of Trichinella spiralis larvae in rats.On the 100th day post-infection(PI),the rats were divided into an acute cold restraint stress(ACRS)group and a non-stressed group.Age-matched untreated rats served as controls.The abdominal withdrawal reflex was used to measure the visceromotor response to colorectal distension(CRD).The expression levels of VGLUT3 in peripheral and central neurons were analyzed by immunofluorescence and western blotting.RESULTS:VGLUT3 expression in the L6S1 dorsal root ganglion cells was significantly higher in the PI group than in the control group(0.32±0.009 vs0.22±0.008,P<0.01),and there was no significant difference in the expression of VGLUT3 between MCdeficient rats and their normal wild-type littermates.Immunofluorescence showed that the expression levels of VGLUT3 in PI+ACRS rats were enhanced in the prefrontal cortex of the brain compared with the control group.CONCLUSION:VGLUT3 is involved in the pathogenesis of visceral hyperalgesia.Coexpression of c-fos,5-hydroxytryptamine and VGLUT3 after CRD was observed in associated neuronal pathways.Increased VGLUT3 induced by transient intestinal infection was found in peripheral nerves,and was independent of MCs.Moreover,the expression of VGLUT3 was enhanced in the prefrontal cortex in rats with induced infection and stress.
