BACKGROUND： Previous studies have demonstrated that postsynaptic density protein-95 （PSD-95） is widely distributed in the central nervous system and is related to the development of the CNS and sensory signal trans...BACKGROUND： Previous studies have demonstrated that postsynaptic density protein-95 （PSD-95） is widely distributed in the central nervous system and is related to the development of the CNS and sensory signal transmission as well as acute or chronic nerve cell death following ischemic brain injury. OBJECTIVE： To semi-quantitatively determine the pathological changes of apoptotic facial neurons and the expression of PSD-95 in the facial nucleus following facial nerve injury of varying extents using immunohistochemical staining methods. DESIGN, TIME AND SETTING： Randomized, controlled animal experiments were performed in the Ultrasonic Institute of the Second Affiliated Hospital of Chongqing University of Medical Sciences from September to December 2007. MATERIALS： Sixty-five healthy, adult, Sprague-Dawley （SD） rats, both male and female, were used for this study. Rabbit anti-rat PSD-95 polyclonal antibody was purchased from Beijing Biosynthesis Biotechnology Co., Ltd. METHODS： SD rats were randomly assigned into a control group with five rats and three injured groups with 20 rats per group. Exposure, clamp and cut for bilateral facial nerve trunks were performed in the rats of the injury groups, and no injury was inflicted on the rats of the control group. MAIN OUTCOME MEASURES; The brainstems of all the rats were excised on days 1, 3, 7, and 14 post injury, and then the facial nuclei were stained with hematoxylin-eosin to observe any pathological changes due to apoptosis in facial neurons. PSD-95 expression in facial nuclei was detected by immunohistochemistry and the number of PSD-95 positive cells was counted under a light microscope. RESULTS： The expression of PSD-95 in the facial nucleus and morphology of the facial neuron within the exposure group had no obvious changes at various points in time tested （P 〉 0.05）. However, the expressions of PSD-95 in the facial nucleus of the clamp group and cut group increased on day 1 post injury （P 〈 0.05）, and showed further increase on day 7 post injury （P 〈 0.01 ）. This did not decrease until day 14 post injury. Facial neuron apoptosis was detected on day 3 post injury and this was even more obvious on day 7 and was maintained to day 14 post injury. The number of cells expressing PSD-95 and displaying severe degrees of facial neuron apoptosis were as follows： cut group 〉 clamp group 〉 exposure group. CONCLUSION： The apoptotic extent of facial neurons and the expression of PSD-95 in apoptotic facial neurons increased with the degree of aggravation of injured severity of facial nerve.展开更多
目的:检测吗啡条件性位置偏爱(conditioned place preference,CPP)重现大鼠海马区突触后致密质-95(PSD-95)的蛋白和DNA表达,观察PSD-95对吗啡成瘾记忆的影响。方法:建立大鼠吗啡CPP模型,自然消退后,通过环境来诱发CPP的重现,应用免疫组...目的:检测吗啡条件性位置偏爱(conditioned place preference,CPP)重现大鼠海马区突触后致密质-95(PSD-95)的蛋白和DNA表达,观察PSD-95对吗啡成瘾记忆的影响。方法:建立大鼠吗啡CPP模型,自然消退后,通过环境来诱发CPP的重现,应用免疫组化和RT-PCR的方法,观察吗啡CPP重现组大鼠海马区PSD-95的表达,并与吗啡CPP消退组、生理盐水对照组进行比较。结果:吗啡CPP环境激发组、吗啡CPP消退组与生理盐水对照组相比,以及吗啡CPP环境激发组与吗啡CPP消退组相比,海马区PSD-95的表达明显降低,差异具有高度显著性(P<0.01)。结论:吗啡CPP重现和消退大鼠海马区PSD-95表达明显降低,海马区PSD-95可能未参与成瘾记忆。展开更多
目的研究脑血流低灌注对大鼠学习记忆、脑组织突触后致密物(postsynaptic density,PSD)超微结构及PSD-95蛋白表达的影响。方法利用双侧颈总动脉永久性结扎制备SD大鼠脑血流低灌注模型。将SD大鼠随机分为假手术组和脑血流低灌注模型组。...目的研究脑血流低灌注对大鼠学习记忆、脑组织突触后致密物(postsynaptic density,PSD)超微结构及PSD-95蛋白表达的影响。方法利用双侧颈总动脉永久性结扎制备SD大鼠脑血流低灌注模型。将SD大鼠随机分为假手术组和脑血流低灌注模型组。采用Morris水迷宫检测大鼠学习记忆情况。采用透射电镜结合Image Pro Plus图像分析系统定量分析大鼠海马突触数量、PSD长度和厚度。采用免疫组化观察大鼠脑组织PSD-95的表达。结果脑血流低灌注大鼠空间参考记忆及工作记忆能力减退,在第2至4天游泳训练时间段,搜寻隐藏平台路径较假手术组明显增加(P<0.05);撤离平台后,在原平台象限游泳时间及路径较假手术组降低,准确穿越原平台所在位置次数减少(P<0.05);当平台移动到第Ⅱ、Ⅳ象限时,搜索移动平台路径较假手术组明显延长(P<0.05)。与假手术组比较,脑血流低灌注大鼠海马突触数量明显减少,PSD长度变短,厚度变薄(P<0.05);海马CA1区、丘脑前内侧核、颞叶皮层PSD-95积分吸光度降低(P<0.05)。结论脑血流低灌注所致学习记忆功能减退与海马突触后致密物结构损伤、PSD-95蛋白表达下降有关。展开更多
目的探讨氟西汀(fluoxetine,FLXT)对创伤后应激障碍(post-traumatic-stress-disorder,PTSD)大鼠记忆及大鼠海马神经元突触后致密物蛋白95(postsynaptic density 95,PSD-95)和突触素Ⅰ(synapsinⅠ)表达的影响。方法采用国际认定的SPS方...目的探讨氟西汀(fluoxetine,FLXT)对创伤后应激障碍(post-traumatic-stress-disorder,PTSD)大鼠记忆及大鼠海马神经元突触后致密物蛋白95(postsynaptic density 95,PSD-95)和突触素Ⅰ(synapsinⅠ)表达的影响。方法采用国际认定的SPS方法刺激大鼠建立PTSD大鼠模型,应用水迷宫实验观察氟西汀对PTSD大鼠学习记忆的影响,采用免疫荧光染色和免疫印迹法检测海马神经元PSD-95和突触素Ⅰ水平。结果 Morris水迷宫前5天的定位航行实验结果显示,SPS刺激后大鼠(PTSD大鼠)找到水下平台的游泳距离和潜伏期比正常组大鼠长,给予氟西汀的PTSD大鼠找到水下平台的距离和潜伏期较未用氟西汀处理的PTSD大鼠缩短。Morris水迷宫第6天空间探索实验结果显示,PTSD大鼠穿越平台的次数和在靶象限花费时间的百分比明显低于正常对照大鼠,而氟西汀可显著增加PTSD大鼠在水迷宫实验中的穿台次数和在靶象限停留的时间百分比。免疫荧光染色和Western Blot检测显示,PTSD大鼠PSD-95和突触素Ⅰ的水平降低,氟西汀干预可抑制PTSD大鼠PSD-95和突触素Ⅰ水平的降低。结论氟西汀可通过抑制PTSD大鼠海马神经元细胞PSD-95和SynapsinⅠ水平的下调,减轻PTSD大鼠空间记忆和学习能力的损伤。展开更多
目的观察小鼠学习记忆能力及其海马区突触功能相关蛋白脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、突触后致密蛋白95(postsynaptic density protein 95,PSD95)及GluA1表达的增龄性变化。方法观察10周龄(青年组)和21...目的观察小鼠学习记忆能力及其海马区突触功能相关蛋白脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、突触后致密蛋白95(postsynaptic density protein 95,PSD95)及GluA1表达的增龄性变化。方法观察10周龄(青年组)和21月龄(老年组)C57BL/6雄性小鼠Morris水迷宫训练和测试表现,并应用Western blot技术检测两组小鼠海马区BDNF、PSD95、GluA1的蛋白表达。结果与青年组相比,老年组小鼠水迷宫测试中的学习记忆能力明显下降(P均<0.05),其海马区总蛋白BDNF、PSD95、GluA1的表达量均显著下降(P均<0.05),海马区膜蛋白GluA1的表达量也明显下降(P<0.05)。结论老年小鼠学习记忆能力下降伴随着海马区突触功能相关蛋白BDNF、PSD95、GluA1表达的一致下降。展开更多
文摘BACKGROUND： Previous studies have demonstrated that postsynaptic density protein-95 （PSD-95） is widely distributed in the central nervous system and is related to the development of the CNS and sensory signal transmission as well as acute or chronic nerve cell death following ischemic brain injury. OBJECTIVE： To semi-quantitatively determine the pathological changes of apoptotic facial neurons and the expression of PSD-95 in the facial nucleus following facial nerve injury of varying extents using immunohistochemical staining methods. DESIGN, TIME AND SETTING： Randomized, controlled animal experiments were performed in the Ultrasonic Institute of the Second Affiliated Hospital of Chongqing University of Medical Sciences from September to December 2007. MATERIALS： Sixty-five healthy, adult, Sprague-Dawley （SD） rats, both male and female, were used for this study. Rabbit anti-rat PSD-95 polyclonal antibody was purchased from Beijing Biosynthesis Biotechnology Co., Ltd. METHODS： SD rats were randomly assigned into a control group with five rats and three injured groups with 20 rats per group. Exposure, clamp and cut for bilateral facial nerve trunks were performed in the rats of the injury groups, and no injury was inflicted on the rats of the control group. MAIN OUTCOME MEASURES; The brainstems of all the rats were excised on days 1, 3, 7, and 14 post injury, and then the facial nuclei were stained with hematoxylin-eosin to observe any pathological changes due to apoptosis in facial neurons. PSD-95 expression in facial nuclei was detected by immunohistochemistry and the number of PSD-95 positive cells was counted under a light microscope. RESULTS： The expression of PSD-95 in the facial nucleus and morphology of the facial neuron within the exposure group had no obvious changes at various points in time tested （P 〉 0.05）. However, the expressions of PSD-95 in the facial nucleus of the clamp group and cut group increased on day 1 post injury （P 〈 0.05）, and showed further increase on day 7 post injury （P 〈 0.01 ）. This did not decrease until day 14 post injury. Facial neuron apoptosis was detected on day 3 post injury and this was even more obvious on day 7 and was maintained to day 14 post injury. The number of cells expressing PSD-95 and displaying severe degrees of facial neuron apoptosis were as follows： cut group 〉 clamp group 〉 exposure group. CONCLUSION： The apoptotic extent of facial neurons and the expression of PSD-95 in apoptotic facial neurons increased with the degree of aggravation of injured severity of facial nerve.
文摘目的:检测吗啡条件性位置偏爱(conditioned place preference,CPP)重现大鼠海马区突触后致密质-95(PSD-95)的蛋白和DNA表达,观察PSD-95对吗啡成瘾记忆的影响。方法:建立大鼠吗啡CPP模型,自然消退后,通过环境来诱发CPP的重现,应用免疫组化和RT-PCR的方法,观察吗啡CPP重现组大鼠海马区PSD-95的表达,并与吗啡CPP消退组、生理盐水对照组进行比较。结果:吗啡CPP环境激发组、吗啡CPP消退组与生理盐水对照组相比,以及吗啡CPP环境激发组与吗啡CPP消退组相比,海马区PSD-95的表达明显降低,差异具有高度显著性(P<0.01)。结论:吗啡CPP重现和消退大鼠海马区PSD-95表达明显降低,海马区PSD-95可能未参与成瘾记忆。
文摘目的研究脑血流低灌注对大鼠学习记忆、脑组织突触后致密物(postsynaptic density,PSD)超微结构及PSD-95蛋白表达的影响。方法利用双侧颈总动脉永久性结扎制备SD大鼠脑血流低灌注模型。将SD大鼠随机分为假手术组和脑血流低灌注模型组。采用Morris水迷宫检测大鼠学习记忆情况。采用透射电镜结合Image Pro Plus图像分析系统定量分析大鼠海马突触数量、PSD长度和厚度。采用免疫组化观察大鼠脑组织PSD-95的表达。结果脑血流低灌注大鼠空间参考记忆及工作记忆能力减退,在第2至4天游泳训练时间段,搜寻隐藏平台路径较假手术组明显增加(P<0.05);撤离平台后,在原平台象限游泳时间及路径较假手术组降低,准确穿越原平台所在位置次数减少(P<0.05);当平台移动到第Ⅱ、Ⅳ象限时,搜索移动平台路径较假手术组明显延长(P<0.05)。与假手术组比较,脑血流低灌注大鼠海马突触数量明显减少,PSD长度变短,厚度变薄(P<0.05);海马CA1区、丘脑前内侧核、颞叶皮层PSD-95积分吸光度降低(P<0.05)。结论脑血流低灌注所致学习记忆功能减退与海马突触后致密物结构损伤、PSD-95蛋白表达下降有关。
文摘目的探讨氟西汀(fluoxetine,FLXT)对创伤后应激障碍(post-traumatic-stress-disorder,PTSD)大鼠记忆及大鼠海马神经元突触后致密物蛋白95(postsynaptic density 95,PSD-95)和突触素Ⅰ(synapsinⅠ)表达的影响。方法采用国际认定的SPS方法刺激大鼠建立PTSD大鼠模型,应用水迷宫实验观察氟西汀对PTSD大鼠学习记忆的影响,采用免疫荧光染色和免疫印迹法检测海马神经元PSD-95和突触素Ⅰ水平。结果 Morris水迷宫前5天的定位航行实验结果显示,SPS刺激后大鼠(PTSD大鼠)找到水下平台的游泳距离和潜伏期比正常组大鼠长,给予氟西汀的PTSD大鼠找到水下平台的距离和潜伏期较未用氟西汀处理的PTSD大鼠缩短。Morris水迷宫第6天空间探索实验结果显示,PTSD大鼠穿越平台的次数和在靶象限花费时间的百分比明显低于正常对照大鼠,而氟西汀可显著增加PTSD大鼠在水迷宫实验中的穿台次数和在靶象限停留的时间百分比。免疫荧光染色和Western Blot检测显示,PTSD大鼠PSD-95和突触素Ⅰ的水平降低,氟西汀干预可抑制PTSD大鼠PSD-95和突触素Ⅰ水平的降低。结论氟西汀可通过抑制PTSD大鼠海马神经元细胞PSD-95和SynapsinⅠ水平的下调,减轻PTSD大鼠空间记忆和学习能力的损伤。
文摘目的观察小鼠学习记忆能力及其海马区突触功能相关蛋白脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、突触后致密蛋白95(postsynaptic density protein 95,PSD95)及GluA1表达的增龄性变化。方法观察10周龄(青年组)和21月龄(老年组)C57BL/6雄性小鼠Morris水迷宫训练和测试表现,并应用Western blot技术检测两组小鼠海马区BDNF、PSD95、GluA1的蛋白表达。结果与青年组相比,老年组小鼠水迷宫测试中的学习记忆能力明显下降(P均<0.05),其海马区总蛋白BDNF、PSD95、GluA1的表达量均显著下降(P均<0.05),海马区膜蛋白GluA1的表达量也明显下降(P<0.05)。结论老年小鼠学习记忆能力下降伴随着海马区突触功能相关蛋白BDNF、PSD95、GluA1表达的一致下降。
