孤独症大鼠前额叶皮质突触相关蛋白的表达

目的 观察孤独症大鼠突触相关蛋白突触素(SYN)、突触后致密蛋白-95(PSD-95)、桥尾蛋白(gephyrin)在前额叶皮质的表达变化,探讨突触相关蛋白在孤独症发病中的作用。方法 采用孕12.5 d(E12.5)一次性腹腔注射丙戊酸钠(600 mg/kg)的孕鼠所产下的子代鼠作为模型组,同样方法注射同等剂量的生理盐水的孕鼠所产下子代鼠作为正常组。通过斜板实验、睁眼实验、三箱实验验证模型是否成功;Western blotting方法对比生后42天两组大鼠突触相关蛋白SYN、PSD-95、gephyrin在前额叶皮质的表达变化。结果 1)成功建立孤独症动物模型:与正常组比较,模型组大鼠生长发育迟缓;社会交往能力异常、对新鲜事物偏好障碍,差异均有统计学意义(P均<0.05);2)Western blotting结果显示:与正常组比较,孤独症大鼠突触相关蛋白SYN、PSD-95表达显著增多(P<0.05),gephyrin蛋白表达显著减少(P<0.05)。结论 孤独症大鼠前额叶皮质突触蛋白表达异常。     

菖蒲郁金汤对高皮质激素血症小鼠海马NF-L及SYP蛋白表达的影响

目的 探讨菖蒲郁金汤拮抗皮质酮诱导海马神经元损伤的作用。方法 将C57BL/6N小鼠随机分为空白组、模型组、氟西汀组、菖蒲郁金汤高剂量组、菖蒲郁金汤中剂量组及菖蒲郁金汤低剂量组。除空白组外,其余各组通过皮质酮诱导高皮质激素血症,予受试药物连续14d,观察小鼠体质量、脾脏质量及胸腺质量变化,采用强迫游泳实验和强迫悬尾实验评价菖蒲郁金汤对小鼠不动时间的影响,Western blot法检测神经微丝轻链蛋白（NF-L）及突触囊泡蛋白（SYP）蛋白表达。结果 菖蒲郁金汤可缓减模型动物体质量、脾脏质量及胸腺质量的降低（P<0.05）;减少强迫游泳实验、强迫悬尾实验小鼠不动时间（P<0.01）;上调NF-L及SYP蛋白的表达（P<0.01）。结论 菖蒲郁金汤通过上调NF-L及SYP的表达,拮抗高皮质激素血症所致的海马神经元损伤,发挥抗抑郁样行为效应。      

大鼠颅脑外伤后突触素P38在中枢神经系统的表达变化

目的观察大鼠颅脑外伤后突触素p38在中枢神经系统表达的变化。方法将SD大鼠分为正常对照组、假手术对照组和损伤组,损伤组用侧位液压冲击法应用不同压力制成颅脑损伤动物模型,于损伤后不同时间点采集标本,采用免疫组织化学技术,检测大鼠损伤灶周边皮层及海马CA1区p38的表达变化,结果采用方差分析和q检验进行统计学分析。结果创伤性颅脑外伤后,损伤灶周边皮层及海马CA1区p38免疫阳性表达增强。结论中枢神经系统损伤后,突触素p38表达增强,且损伤越重,表达增强越明显。     

累加电针对坐骨神经痛大鼠海马及下丘脑突触素表达的影响*

目的：观察电针（EA）镇痛的累积效应,及其与海马及下丘脑突触素（SYN）表达的关系。方法：Wistar雌鼠70只,随机分为正常对照组（n=10）,慢性压迫性损伤（CCI）组（n=30）,去卵巢（OVX）＋CCI组n=30）。后2组又各分为不电针、EA2d和2周组,每组10例。OVX动物去除双侧卵巢,每天颈背部皮下注射D-半乳糖,7周后水迷宫法测试动物学习记忆能力。结扎坐骨神经造成CCI慢性痛模型。电针双侧“足三里”-“阳陵泉”穴,1次／d,不同组分别电针2d、2周。辐射热照射测定大鼠缩腿潜伏期,以两足的差值作为痛敏分数（HAS）。结果：CCI后,动物HAS明显增大。CCI＋EA2周组HAS的绝对值显著低于CCI组、CCI＋EA2d组（P〈O．05）;显著低于OVX＋CCI＋EA2周组（尸〈O．05）。与CCI组比较．CCI＋EA2周组下丘脑PVN、海马CA1区SYN积分灰度值明显较低（表达上调;P〈0．05）;而CCI＋EA2d组无明显变化。OVX＋CCI＋EA2周组两脑区SYN积分灰度值明显低于OVX＋CCI组（P〈0．05）,而显著高于CCI＋EA2周组（P〈0．05）。与CCI＋EA2周组比较,OVX＋CCI＋EA2周组SYN表达的上调程度低（P〈0．05）。结论：重复电针有累积镇痛效应：海马及下丘脑SYN表达上调与针刺累积效应密切相关;神经记忆力的减退在可减弱针刺镇痛的累积效应。     

苯甲酸雌二醇对去卵巢大鼠海马突触素和钙结合蛋白免疫活性的影响

目的　探讨苯甲酸雌二醇对大鼠海马CA1区、齿状回突触素(synaptophysin, Syn)及钙结合蛋白(calbindinD28k, CaBP, parvalbumin, PV)的调节作用。方法　用大鼠卵巢切除(OVX)动物模型,用免疫组化结合图象分析的方法。结果　OVX组大鼠海马CA1区及齿状回突触素、PV,CaBP积分光密度及面积密度均显著低于对照组,补充苯甲酸雌二醇35 d,这3项指标的积分光密度及面积密度均告恢复,与OVX组相比,呈显著性差异。结论　苯甲酸雌二醇可能通过调节海马、齿状回神经元突触密度,为改善学习记忆功能,缓解AD病人的症状提供结构基础,并通过维持胞内Ca²⁺的恒定达到保护神经元的目的。     

Expression of synaptic proteins in the developing rat cerebellum following ionizing radiation

Various proteins regulating neurotransmission release and synaptic vesicle exocytosis have been implicated in axonal elongation and synaptic maturation. In the present study, immunohistochemistry to the presynaptic membrane proteins syntaxin-I and synaptosomal-associated protein of 25 kDa (SNAP-25) synaptic vesicle-associated proteins synaptophysin and synapsin-I and the neuronal maturation and axonal growth-related protein GAP-43, has been carried out in the normal developing cerebellum and following a single dose of ionizing radiation (2 Gy gamma-rays) at postnatal day 1. Our aim has been to learn about the morphological and possible functional modalities that occur during the progression of neuronal connectivity in normal and abnormal development. Expression of all these proteins is associated with the arrival of afferents in the subcortical white matter and with the maturation of the internal granule cell layer and molecular layer during normal development. In addition, SNAP-25 and GAP-43 are strongly expressed in granule cells of the external granule cell layer, thus suggesting that these proteins are involved in cell elongation of granule cells. Apoptosis appears at 3 h and peaks at 6 h following ionizing radiation. Radiation-induced apoptosis in the external granule cell layer produces a transient decrease in the expression of SNAP-25 and GAP-43 in the external granule cell layer. The external granule cell layer recovers at 48 h and external granule cells of proliferating cells also express SNAP-25 and GAP-43, thus indicating that proliferating cells in this layer are equipped with proteins involved in cell elongation. Furthermore, expression of synaptophysin, synapsin-I, syntaxin-I and SNAP-25 is the same in the cerebellum of irradiated and normal rats from this time to adulthood (3 months). These results point to the likelihood that recovery of the cerebellar cortex occurs following a single exposure of ionizing radiation during postnatal development.     

补阳还五汤对脑缺血再灌注大鼠恢复期突触可塑性的影响

目的:探讨补阳还五汤在脑缺血再灌注大鼠恢复期提高突触可塑性的机制研究。方法:建立大脑中动脉栓塞(MCAO)大鼠模型,随机分为假手术组、模型组、补阳还五汤组、补阳还五汤联合缝隙连接蛋白43(Cx43)抑制剂(Gap26)组,补阳还五汤每天灌胃2次(16 g·kg^-1),Gap26于术后第3天腹腔注射,每天1次(25μg·kg^-1);7 d后取材,采用透射电镜观察缺血侧海马突触和缝隙连接超微结构的改变,运用蛋白免疫印迹法(Western blot)和免疫荧光检测缺血侧海马突触素(SYN),生长相关蛋白-43(GAP-43)的表达。结果:电镜下观察到假手术组突触结构完整、清晰,突触数量多,缝隙连接结构清晰;模型组缺血侧海马突触结构溶解,突触数量减少,缝隙连接消失,存在较大间隙,与假手术组比较,SYN,GAP-43的表达明显增高(P<0.05,P<0.01);补阳还五汤组缺血侧海马突触结构较清晰,突触数量增多,缝隙连接结构较完整,与模型组比较,SYN,GAP-43的表达明显增强(P<0.05,P<0.01);而联合使用Gap26后缺血侧海马突触数量较补阳还五汤组减少,仅可见少量结构完整的缝隙连接,补阳还五汤增强SYN,GAP-43的作用被明显抑制(P<0.05,P<0.01)。结论:补阳还五汤可提高脑缺血再灌注恢复期缺血侧海马突触可塑性,其机制可能与增加Cx43的表达促进对SYN,GAP-43的干预有关。     

成人髓母细胞瘤患者的预后影响因素分析

目的探讨成人髓母细胞瘤(MB)患者性别、肿瘤部位、免疫组化指标、是否全切、辅助放化疗等临床、病理因素对预后的影响。方法回顾性分析25例成人髓母细胞瘤患者的临床资料,并对患者的预后进行电话随访。使用Kaplan-Meier曲线和Log-rank检验进行生存分析,寻找影响预后的因素。结果本组患者中,男性14例(56.0%),女性11例(44.0%),平均年龄为(25.1±5.9)岁。其中,12例患者(48.0%)的肿瘤位于小脑内,13例患者(52.0%)肿瘤突入四脑室;20例患者(80.0%)实现了肿瘤全切;23例患者(92.0%)术后进行了放疗;13例患者(52.0%)术后行化疗。本组患者的5年无进展生存和整体生存率分别为(65.9±10.6)%和(67.5±15.6)%。辅助放疗(P<0.001)和突触素阳性表达(P=0.017)与较好的5年无进展生存相关。在术后行辅助放疗的基础上,再行辅助化疗并无确切疗效(P=0.202)。结论成人髓母细胞瘤患者术后行辅助放疗是必需的,是否行辅助化疗尚存争议。辅助放疗和突触素阳性表达与较好的预后相关。免疫组化指标对评估成人髓母细胞瘤患者的预后有价值。     

Synaptophysin – a common constituent of presumptive secretory microvesicles in the mammalian pinealocyte: A study of rat and gerbil pineal glands

Recent studies have established that pinealocytes of the mammalian pineal gland contain marker molecules of neuroendocrine cells or paraneurons like the synaptic vesicle-associated protein synaptophysin (p38). The objective of this study was to identify the subcellular synaptophysin-positive compartment and to characterize in detail the intracellular distribution of this protein in rat and gerbil pinealocytes. An analysis of serial semithin sections of plastic-embedded pineals immunostained for synaptophysin, including computer-assisted optical density measurements of synaptophysin immunoreactivities, demonstrated unequivocally that synaptosphysin was highly concentrated in dilated process terminals of the pinealocytes. More than 75% of these process terminals were found to border or lie within the pericapillary space. At the ultrastructural level, they contained accumulations of small clear vesicles of variable size that turned out to be the site of synaptosphysin immunoreactivity when immunogold staining was performed. In addition, microvesicles surrounding synaptic ribbons were also immunolabeled. Hence, the pinealocyte is the first neuroendocrine cell type that has now been shown to concentrate synaptophysin-positive microvesicles in perivascular process endings. This observation lends strong support to the hypothesis that small clear vesicles in neuroendocrine cells in general, and in pinealocytes in particular, serve secretory functions. The quantitative analysis of completely sectioned process endings revealed that the microvesicles outnumber by far the amount of dense core vesicles and therefore cannot arise by endocytosis of dense core vesicle membranes. Thus, small synaptic-like vesicles probably constitute an independent secretory pathway of the paraneuronal pinealocytes. In the present study, we could also establish the absence of immunoreactivity for synapsin I (belonging to a family of neuron-specific nerve terminal phosphorproteins) from pinealocytes. Synapsin I immunoreactivity was only detectable in intrapineal nerve terminals and varicosities. Taken together, the immunostaining patterns of the pineal gland obtained with antibodies directed against synaptic vesicle-associated proteins render the mammalian pinealocyte a very special type of neuroendocrine cell or paraneuron rather than a “classic” neuron. © 1993 Wiley-Liss, Inc.