癫痫大鼠内嗅皮质神经元及GFAP免疫反应活性的变化

目的观察癫癎敏感大鼠内嗅皮质神经元及GFAP免疫反应活性的变化.方法用红藻氨酸(KA)10 mg·kg-1诱发大鼠出现癫癎发作.1个月后,用免疫组化及硫瑾染色的方法观察大鼠内嗅质层(EC)神经元及神经胶质原纤维酸性蛋白(GFAP)免疫反应阳性细胞数目的变化.结果发现EC的第3层细胞有大量的神经元损失,伴有GFAP免疫反应活性增强.结论这一现象可能与动物癫癎敏感性长期增强有关.     

癫痫敏感大鼠内嗅皮层GABA及Kappa受体免疫反应活性的变化

目的研究癫痫敏感大鼠内嗅皮层(EC)GABA及Kappa受体免疫反应活性的变化。方法用惊厥剂量(10mg／kg．sc)的红藻氨酸(Kainic acid,KA)制备癫痫发作敏感大鼠,用免疫组化方法观察内嗅皮层(EC)GA- BA和Kappa受体免疫反应活性的变化。结果与对照组比较,KA后4周,癫痫敏感动物EC部位GABA免疫反应活性(GABA-IR)及Kappa受体免疫反应活性(Kappa-R-IR)均明显减少(P<0．05)。结论 EC处GABA-IR及Kap- pa-R-IR的长期下调很可能是大鼠癫痫发作敏感性长期增强的重要原因。     

癫痫大鼠脑内白细胞介素6的表达特征

有临床资料显示，有癫痫病史或癫痫发作的患者脑脊液和血浆白细胞介素6(IL-6)浓度升高，动物实验也证明，在各种癫痫发作模型中脑内IL-6表达增加。我们试图通过研究红藻氨酸(KA)诱导癫痫发作的动物模型相关脑区中IL-6的表达特征及其与c—Fos表达的比较，进一步探讨IL-6在癫痫发作中的意义及其细胞学来源。     

红藻氨酸致痫大鼠海马Fos和GFAP的共同表达

目的 研究红藻氨酸（kainic acid,KA)诱导大鼠癫痫发作后海马（hippocampus,HI)内神经元和星形胶质细胞的时空效应性反应变化。方法 大鼠侧脑室内注射KA，用抗即刻早期基因Fos蛋白和抗胶质原纤维酸性蛋白（GFAP）的双重免疫荧光组织化学方法结合激光共聚焦显微镜技术，显示痫性发作后HI同一部位内反应性神经元与星形胶质细胞的分布。结果 KA诱导大鼠癫痫发作，HI内的Fos阳性神经元和GFAP阳性星形胶质细胞明显增多。两分布范围基本一致，且癫痫诱发30min后GFAP开始增多，1h达高峰；1h后Fos阳性产物开始增多；2h达高峰；部分Fos阳性神经元周围有GFAP免疫反应产物包绕，显示反应性神经元（Fos阳性）与反应性星形胶质细胞（GFAP阳性）之间关系密切。结论 HI内的神经元和星形胶质细胞与癫痫发作直接相关且存在相互关系。可能共同参与癫痫的发生及其调节。     

红藻氨酸诱导大鼠癫痫发作脑组织HSP70表达的免疫组化？…

目的探讨热休克蛋白（ＨＳＰ）７０在癫痫发作后的表达及作用。方法 用免疫组化免疫免印迹分析法观察红藻氨酸诱导大鼠癫痫发作后ＨＳＰ７０的表达及其动态演变过程。     

甘珀酸对癫痫大鼠前脑connexin-32和GFAP表达研究

目的研究大鼠在脑室埋管注射甘珀酸(CBX)预处理后戊四氮导致癫痫发作时前脑内胶质纤维酸性蛋白(GFAP)和连接蛋白-32(Cx32)表达及其相互关系. 方法动物分为生理盐水(NS)对照组(NS组)、CBX预处理对照组(CBX组)、戊四氮(PTZ)致痫组(PTZ组)、CBX预处理后再PTZ致痫组(CBX PTZ组),应用免疫荧光组织化学双重标记显示GFAP和Cx32在前脑的表达及其相互关系.结果 CBX PTZ组的大鼠癫痫发作的行为表现比PTZ组显著加重,星形胶质细胞GFAP的表达也比PTZ组明显升高,值得注意的是CBX组的星形胶质细胞GFAP的表达比NS组明显升高.Cx32在PTZ引起的癫痫大鼠的大脑皮层、海马和杏仁核内是增加的,而CBX预处理后的癫痫模型中Cx32却降低了.在Cx32与GFAP双标的切片上发现,Cx32与GFAP阳性的星形胶质细胞非常接近,在CBX预处理后的癫痫模型中GFAP阳性的星形胶质细胞显著增加的同时Cx32却明显降低. 结论在整体动物CBX预处理可以导致癫痫发作增强,这可能与星形胶质细胞增生及其Cx32表达降低有关.     

红藻氨酸诱导性癫痫发作大鼠海马cNOS及iNOS的变化及作用

目的研究组成型一氧化氮合酶（ｃＮＯＳ）及诱导型一氧化氮合酶（ｉＮＯＳ）在红藻氨酸（ＫＡ）诱导癫痫发作中的变化及作用。方法采用免疫组织化学方法显示ｃＮＯＳ及ｉＮＯＳ的变化;Ｎｉｓｓｌ染色显示神经元的损害。结果ＫＡ３０分钟ｃＮＯＳ较对照组明显增加（Ｐ＜０．０５）,随后下降至正常水平;ＫＡ诱导２小时ｉＮＯＳ明显升高,以ＣＡ１区为著,至ＫＡ６小时达高峰,然后在高水平缓慢下降;Ｎｉｓｓｌ染色神经元变性坏死ＣＡ３＝齿状回＞ＣＡ１。结论ＫＡ诱导癫痫发作导致海马ｃＮＯＳ及ｉＮＯＳ表达增多,神经元的坏死与ＮＯＳ表达增多无明显关系。     

红藻氨酸诱导癫痫发作大鼠海马EphA5受体及其配体ephrinA3基因表达变化的研究

目的研究在红藻氨酸(Kainic acid,KA)诱导的损伤型颞叶癫痫(Mesial temporal lobe epilepsy,MTLE)的大鼠海马中,轴突导向因子EphA5受体及其配体ephrinA3基因表达的变化,探讨EphA5/ephrinA3与癫痫后海马兴奋性神经网络形成的作用和关系。方法侧脑室内微量注射KA,建立KA诱导的成年大鼠MTLE模型,用原位杂交法检测癫痫发作1d、1周、2周、3周、4周大鼠海马内EphA5/ephrinA3 mRNA的表达,定量分析表达的动态变化。结果EphA5/ephrinA3 mRNA于癫痫发作后1周,在海马齿状回颗粒细胞层和CA_3区锥体细胞层开始增强,2周达到高峰,4周恢复接近对照组水平。结论在KA所致的癫痫持续状态(Status epilepsy,SE)中,海马神经元通过增强EphA5/ephrinA3 mRNA的表达。调控MTLE大鼠海马内苔藓纤维和突触的重建,是癫痫后海马新的稳定的异常兴奋性神经网络形成的可能机制。     

红藻氨酸诱导癫痫大鼠前脑CX32的表达及辛醇干预的影响

目的 观察红藻氨酸(kainic acid,KA)诱导癫痫大鼠前脑缝隙连接蛋白32(connexin 32,CX32)的表达及辛醇干预的影响.方法 用免疫荧光法检测癫痫发作后各时间点大鼠皮质及海马CX32阳性细胞表达.同时观察致痫前给予辛醇干预对大鼠皮质及海马CX32阳性细胞表达的影响.结果 KA致痫组大鼠皮层及海马CX32阳性细胞在各时程明显高于正常对照组(P<0.05),并且随时程延长呈增加趋势,7d达高峰,在海马中的变化比皮层明显,但无统计学意义(P>0.05).辛醇干预组大鼠皮层及海马CX32阳性细胞在各相应时程明显低于KA致痫组(P<0.01).结论 CX32组成缝隙连接(gap junction,GJ)在癫痫的发生发展过程中起重要作用,辛醇可以减少癫痫大鼠CX32表达,降低癫痫敏感性,实现脑保护作用.     

红藻氨酸诱导的大鼠癫痫模型中海马神经元caspase-9活性的变化

目的 :观察红藻氨酸 (kainicacid ,KA)诱导的癫大鼠海马神经元caspase 9活性的变化。方法 :应用KA腹腔注射诱导大鼠癫模型 ,检测癫大鼠海马神经元caspase 9的活性。caspase 9活性的测定采用荧光底物分析法。结果 :注射KA后 1h大鼠海马神经元caspase 9活性开始升高 ,但与对照组相比 ,无统计学差异 ;注射KA后 3h大鼠海马神经元caspase 9活性达到高峰 ,与对照组相比P <0 0 5 ;然后开始下降 ,于注射KA后 12h接近于正常对照组水平。结论 :在KA腹腔注射诱导的大鼠癫模型中 ,海马神经元caspase 9活性仅在早期表达升高 ,提示caspase 9活性升高可能启动了癫大鼠海马神经元的损伤     

Specific monoclonal antibodies to glial fibrillary acidic protein (GFAP)

Anti-glial fibrillary acidic protein (GFAP) monoclonal antibody has been obtained by fusing SP2/O myeloma cells with splenic cells from mice immunized with human GFAP. It belongs to the IgG1 class and it recognizes an epitope on GFAP which is shared by each fragment of the protein. Immunohistological studies show that the epitope characterized is absolutely specific for GFAP.     

大鼠脑出血急性期延髓内脏带内胶质原纤维酸性蛋白表达的变化

观察脑出血急性期大鼠延髓内脏带 ( MVZ)内星形胶质细胞的反应。以尾壳核局部注射胶原酶制作脑出血模型 ,用抗星形胶质细胞特异性标识物胶质原纤维酸性蛋白 ( GFAP)的免疫细胞化学方法 ,研究脑出血后 MVZ内星形胶质细胞的变化。发现脑出血后 4h GFAP阳性细胞数量增多、胞体增大、突起伸长 ,在MVZ形成明显弧形带状分布 ,尤以 MVZ背内侧区、中间带及腹外侧区明显。提示 MVZ内星形胶质细胞可能参与了脑出血后的病理生理过程。     

Effects of Pulsed Magnetic Stimulation on GFAP Levels in Cultured Astrocytes

The present study evaluates the physiological effects of magnetic stimulation on astrocyte cultures. Cell cultures were exposed to pulsed magnetic stimulation (10 Hz, 10 sec) at the following levels: 0.10 tesla (T; Group A); 0.21 T (Group B); 0.42 T (Group C); and 0.63 T (Group D). Glial fibrillary acidic protein (GFAP) levels from immunoblots, total protein concentrations, and cellular morphology were analyzed at 0, 1, 3, 5, 7, 13, and 20 days poststimulation. Significantly higher GFAP levels were observed in Group D at day 3 (P = 0.0114). The change was transient as the GFAP levels quickly returned to control levels by day 5. No other significant changes in GFAP levels were observed. In comparison to control protein levels at day 0, concentrations from Groups B, C, and D were significantly lower (P < 0.006), whereas at day 3, Groups C and D were significantly higher (P < 0.02). Differences in astrocyte morphology due to magnetic stimulation were not observed. This study demonstrated that high intensity magnetic stimulation for only 10 sec induced a transient biological response. J. Neurosci. Res. 55:238–244, 1999. Published 1999 Wiley‐Liss, Inc.     

Chronic Ethanol-Induced Glial Fibrillary Acidic Protein (GFAP) Immunoreactivity: An Immunocytochemical Observation in Various Regions of Adult Rat Brain

In the present study, the effects of chronic ethanol (ETOH) treatment on the glial fibrillary acidic protein (GFAP) immunoreactivity was investigated in adult rat brains. ETOH were administered as increasing concentrations of 2.4%–7.2% (v/v) gradually for 21 days. Immunocytochemistry revealed that chronic-ETOH treatment increased synthesis of GFAP. The increase in the diameter and the number of GFAP (+) cells were statistically significant compared with the control group (p <. 05). An increase of GFAP immunoreactivity was evident in various white matter and gray matter structures. We concluded that functional astrocytic cells responded to chronic ETOH exposure by increasing the synthesis of GFAP.     

Neuronal modulation of Schwann cell glial fibrillary acidic protein (GFAP)

Adult rat sciatic nerves contain cytoskeletal peptides that resemble CNS glial fibrillary acidic protein (GFAP) in immunoreactivity and molecular weight. Immunohistological examination of teased nerve fascicles indicated that these peptides are expressed selectively by Schwann cells related to small axons. Radiolabelled mouse and rat CNS GFAP cDNA probes hybridized with a single, 2.7 kb RNA band in Northern blots prepared from total RNA from both rat sciatic nerve and rat brain. Sciatic nerve GFAP mRNA was detectable by this means in adult, 2 month, or 21 day postnatal rats, but not in 3,6, or 10 day postnatal rats. Sciatic nerve transection caused a marked reduction in the level of GFAP mRNA in the axotomized distal stump. We conclude that Schwann cell synthesis of GFAP is developmentally regulated and that Schwann cells, unlike astroglia, require continued trophic input from small axons in order to express GFAP.     

Glial fibrillary acidic protein (GFAP) in rat brain tumors transplacentally induced by ethylnitrosourea (ENU)

The immunohistochemical distribution of glial fibrillary acidic protein (GFAP) in neoplastic lesions induced in the rat by ENU is reported. GFAP was present in hypertrophic reactive astrocytes, which were numerous in early neoplastic proliferations, in microtumors of the white matter, and in those collected at the periphery of large tumors. They were absent in cortical oligodendroglial foci and microtumors. No GFAP-positive cells were observed in hyperplasias of the white matter: astrocyte-like cells of large tumors were GFAP-negative. The significance of reactive astrocytes and the problem of the astrocytic component in transplacental ENU tumors are discussed.     

Water maze experience and prenatal choline supplementation differentially promote long‐term hippocampal recovery from seizures in adulthood

Status epilepticus (SE) in adulthood dramatically alters the hippocampus and produces spatial learning and memory deficits. Some factors, like environmental enrichment and exercise, may promote functional recovery from SE. Prenatal choline supplementation (SUP) also protects against spatial memory deficits observed shortly after SE in adulthood, and we have previously reported that SUP attenuates the neuropathological response to SE in the adult hippocampus just 16 days after SE. It is unknown whether SUP can ameliorate longer‐term cognitive and neuropathological consequences of SE, whether repeatedly engaging the injured hippocampus in a cognitive task might facilitate recovery from SE, and whether our prophylactic prenatal dietary treatment would enable the injured hippocampus to more effectively benefit from cognitive rehabilitation. To address these issues, adult offspring from rat dams that received either a control (CON) or SUP diet on embryonic days 12–17 first received training on a place learning water maze task (WM) and were then administered saline or kainic acid (KA) to induce SE. Rats then either remained in their home cage, or received three additional WM sessions at 3, 6.5, and 10 weeks after SE to test spatial learning and memory retention. Eleven weeks after SE, the brains were analyzed for several hippocampal markers known to be altered by SE. SUP attenuated SE‐induced spatial learning deficits and completely rescued spatial memory retention by 10 weeks post‐SE. Repeated WM experience prevented SE‐induced declines in glutamic acid decarboxylase (GAD) and dentate gyrus neurogenesis, and attenuated increased glial fibrilary acidic protein (GFAP) levels. Remarkably, SUP alone was similarly protective to an even greater extent, and SUP rats that were water maze trained after SE showed reduced hilar migration of newborn neurons. These findings suggest that prophylactic SUP is protective against the long‐term cognitive and neuropathological effects of KA‐induced SE, and that rehabilitative cognitive enrichment may be partially beneficial. © 2010 Wiley‐Liss, Inc.     

Glial fibrillary acidic protein (GFAP) in ependymal cells during development. An immunocytochemical study

Human ependymal cells show positive immunostaining for glial fibrillary acidic protein (GFAP) at one stage of the fetal development. The reaction seems to coincide with maturation of the epithelial layer and development of cilia. Two types of reactive cells are present: epithelial and tanycytes. The GFAP-positive reaction in both these cells is transient, appearing at different times and with different patterns in the various regions of the ventricular system. In order to explain the presence of detectable GFAP in developing ependymal cells and its absence in mature cells, it is proposed that either the synthesis of detectable amounts of GFAP occurs only at a stage of ependymal cell maturation, or that the intermediate filaments assembled in developing ependymal cells are antigenically distinct from those of the mature cells. The present findings indicate that tanycytes are not an immature form of ependymal cells but that they develop parallel to the epithelial cells. The role of the tanycytes remains obscure, but it is suggested that they are not related to radial glia.