衰老晚期海马CA1区胶质细胞超微结构改变

选用ＳＤ大白鼠２０只分青年组（３个月）、老年组（３４￣３６个月），应用透视电镜观察比较海马ＣＡ１区胶质细胞的超微结构变化，结果如下：和青年组相比，老年组海马ＣＡ１区胶质细胞胞体及突起出现反应性增生，肥大、包绕神经元及终末，胶质细胞内的线粒体数量减少，肿胀、空洞化线粒体增多，粗面内质网稀疏，脂竭素沉积严重等改变，表现为胶质细胞严重退变，表明衰老晚期海马ＣＡ１区胶质细胞出现对神经元退变代偿的反应性增大     

脑内移植时星状胶质细胞的变化

本文用胶质纤维酸性蛋白免疫组织化学方法研究了帕金森氏病模型鼠、老年性痴呆模型鼠及皮质损伤鼠等在脑移植２～３个月后移植区及其与宿主交界面的胶质细胞变化。结果发现不同动物模型的胚脑移植区及其与宿主交界面的胶质变化基本相同．Ｎｉｓｓｌ染色切片上可见移植区和宿主之间有直径小于８μｍ的小细胞集聚。胶质纤维酸性蛋白免疫组织化学法发现移植区和宿主脑交界面有深染的阳性纤维及细胞或交界面形成一包膜状结构；还有的在交界面无明显的胶质纤维酸性蛋白阳性纤维分隔，呈现两者相互融合的趋势．移植区内可见两种胶质细胞，一种与正常胶质细胞近似，另一种为胞体大、突起粗且深染的反应性胶质细胞；移植区内的血管周围也见有反应性胶质细胞包绕。本研究表明移植也可引起胶质反应，并且移植区内反应性胶质细胞可能对植入的神经元提供其生存所需要的营养物质。     

大鼠脑缺血长期存活后海马可逆性的研究

本实验试图进一步探讨经长期存活后，脑缺血及不全脑缺血大鼠海马的可逆性变化，并验证它们之间的关系。脑缺血模型采用ｐｕｌｓｉｎｅｌｌｉ四血管结扎及其发迹法，苔藓纤维显示采用Ｔｉｍｍ染色法。结果显示，大鼠脑缺血２０ｍｉｎ，现地存活９０ｄ后，海马ＣＡ１区细胞几乎全剖丧失，ＣＡ２／ＣＡ３也出现严重的细胞消亡，ＣＡ１区明显萎缩，多数伴有苔伯枝抽芽，同期不全脑缺血大鼠海马来未见或仅见局限于ＣＡ１的部分细胞丢失，     

激光损伤视网膜后神经元凋亡及GFAP表达的研究

目的 观察兔视网膜激光损伤后神经元细胞有无凋亡改变及视网膜Ｍｕｌｌｅｒ细胞胶质纤维酸性蛋白（ＧＦＡＰ）的表达变化。方法 应用末脱氧核苷酸转移酶介导的ｄＵＴＰ－Ｘ切口末标记法（ＴＵＮＥＬ法）标记凋亡细胞。应用免疫组化染色显示视网膜Ｍｕｌｌｅｒ细胞ＧＦＡＰ表示。结果伤后６ｈ、１、３、７ｄ视网膜各层可见散在分布的ＴＵＮＥＬ阳性凋亡细胞,尤以外核层多见,伤后３ｄ,视网膜可见Ｍｕｌｌｅｒ细胞ＧＦＡＰ表达;伤     

碱性成纤维细胞生长因子的自分泌与反应性星形细胞胶质化

目的观察碱性成纤维细胞生长因子（ｂＦＧＦ）的自分泌现象及其与反应性星形胶质化的关系。方法在大鼠星形胶质细胞（ＡＳ）原代培养的机械损伤模型上，以免疫细胞化学及原位杂交技术检测ｂＦＧＦ、ＰＣＮＡ、ＧＦＡＰ和ＧＦＡＰ－ｍＲＮＡ的动态变化。结果（１）损伤边缘的ＡＳ于伤后２小时起表达ｂＦＧＦ，１２小时达高峰，２天后回落；（２）损伤边缘的ＡＳ于伤后６小时起表达ＧＦＡＰ－ｍＲＮＡ，１天达高峰，２天后回落；（３）伤后１天起，ＧＦＡＰ表达明显增强，ＡＳ胞体肥大并向损伤区伸出粗大突起，２天时ＧＦＡＰ表达至高峰；（４）损伤边缘的部分ＡＳ从伤后２小时起表达ＰＣＮＡ，１天时达到高峰。结论（１）受损的ＡＳ能自分泌ｂＦＧＦ，这是ＡＳ受损后的早期反应之一，具有促进反应性星形胶质化的作用；（２）反应性星形胶质化时，ＧＦＡＰ表达增强是转录水平上ＧＦＡＰ－ｍＲＮＡ增加的结果；（３）反应性星形胶质化以ＡＳ肥大为主，增生为辅。     

雌三醇对胚胎小鼠脑发育的影响

用免疫吸附及免疫组织化学方法研究了在妊娠期给予雌三醇对小鼠胚胎脑发育的影响。免疫定量结果表明：在小鼠生后１４ ｄ,雌三醇可增加神经元特异性标记物 神经元特异性烯醇化酶及神经丝的表达,而神经胶质细胞特异性标记物 神经胶质纤维酸性蛋白不受影响。免疫组织化学反应结果表明：雌三醇增加了位于视束前区,下丘脑视交叉上核、腹内侧核以及杏仁核等结构神经元的神经元特异性烯醇化酶的阳性反应强度,也增加了位于视束前外侧核神经元的神经丝反应强度。雌三醇除了明显减少了海马神经胶质细胞的神经胶质纤维酸性蛋白的表达外,在脑的其它区域对该抗原的反应性未见影响。本研究结果表明：妊娠期给予雌三醇可影响胚胎脑“性中枢”（ＰＯＡ ＡＭ ＢＨ ＡＭ 功能复合体）的发育。     

马桑内酯所致慢性癫痫大鼠海马星形胶质细胞胶质纤维酸性蛋 …

用免疫组织化学方法研究了系统应用马桑内酯所致的慢性癫痫大鼠海马中星表胶质细胞伯胶质纤维酸性蛋白的表达。结果证明，整个海马的胶质纤维酸性蛋白免疫反应明显增强，并可见阳性细胞增生，胞体肥大，尤以齿状回门区和海分子腔隙层及始层为甚，此外，本实验还发现海胶质纤维酸性蛋白的阳性反应随发作后不同时间间隔（２ｈ～９ｄ）而不同，而直至发作后９ｄ胶质纤维酸性蛋白阳性反应程度仍高于对照组，此结果表明，马桑内酯所致癫痫     

不同年龄大鼠大脑皮质损伤后的星形胶质细胞反应

对幼年（３月龄）、成年（１２月龄）与老年（２７月龄）三组雄性Ｗｉｓｔａｒ大鼠的躯感皮质进行针刺后，在１～２１ｄ时间内用光镜与电镜观察损伤后的星形胶质细胞反应，探讨年龄因素对星形胶质细胞反应的影响。结果表明：反应性星形细胞在水肿、细胞分裂、吞噬、肥大与胶质界膜形成等方面均表现出程度上和发生时间上的年龄差异。幼年大鼠的星形细胞不仅水肿更为显著且出现更早，并且表现出向变性与最后死亡以及向代偿性合成增强两个方向发展、三组动物的星形细胞分裂活动以术后３ｄ最明显，尤以幼年组更多见，并出现活跃的吞噬现象．     

脑啡肽对早期反应性胶质增生形态及其神经营养功能的影响

目的　研究脑啡肽对早期反应性胶质细胞形态、增殖及神经营养功能的影响。方法　在相差显微镜下观察反应性胶质细胞形态,３Ｈ－ＴｄＲ掺入实验检测反应性胶质细胞增殖,以Ｇｒｉｅｓｓ反应测定亚硝酸盐含量表示一氧化氮（ＮＯ）量,原位杂交实验检测生长相关蛋白（ｇｒｏｗ ｔｈ ａｓｓｏｃｉａｔｅｄ ｐｒｏｔｅｉｎ,ＧＡＰ－４３）ｍ ＲＮＡ 的表达。结果　脑啡肽能促进损伤边缘的胶质细胞向损伤裸露区伸出宽大扁平的突起,增强反应性胶质细胞３Ｈ－ＴｄＲ的掺入（Ｐ＜０．０５）,抑制其ＮＯ的生成（Ｐ＜ ０．０５）;经脑啡肽处理的反应性胶质细胞能增强神经元ＧＡＰ－４３ ｍ ＲＮＡ 的表达（Ｐ＜０．０５）。结论　脑啡肽增强反应性胶质增生的同时抑制了反应性胶质细胞ＮＯ 的生成而增强反应性胶质细胞的神经营养功能。     

人视网膜星形胶质细胞发育及与血管前体细胞的关系

目的：研究人视网星菜胶质细胞的发育及与血管前体细胞的关系。材料和方法：收集１３４例发育各期胎儿视网膜和４例成人视网膜、石蜡包埋切片整装铺片，四种抗体免疫组分染色，光镜观察，结果：星形胶质细胞分为三种：（１）Ｓ－１００（＋）／胶质纤维酸性蛋白（ＧＦＡＰ）（＋）的双极形星形胶质细胞，视盘进入视网膜，与跟随其后的纤连蛋白（Ｆｎ）（＋）的血管前体细胞接触并相伴向锯齿缘迁移，足月后和成人此类星形胶质细胞主要     

Axon growth failure following corpus callosum lesions precedes glial reaction in perinatal rats

The present study compares the glial reactivity and the axon growth following corpus callosum (CC) lesions, in perinatal rats. Lesions were performed on fetal (E17 to E20) and early postnatal (P0 and P2) rats. The reactive glia and the presence of neural fibers were detected by immunohistochemical staining of glial fibrillary acidic protein (GFAP) and neurofilament protein (NFP), respectively. The callosal axons failed (at least in part) to penetrate the lesioned area already after E18 lesions, and the lesioned area was always impenetrable for axons after E20 and P0 lesions. In these cases, the lesioned part of the CC was completely or nearly devoid of GFAP as well as NFP. The distributions of the immunopositivities to GFAP and NFP also coincided with each other, both in the intact part of the CC and along the alternative courses of the callosal axons. GFAP-immunopositive reactive glia accompanied to the deficiency of NFP-immunostaining only when animals were lesioned at P2. Nestin immunostaining revealed astrocytes or their precursors already at P0, but reactive glia were detected only after P2 lesions, as with immunostaining to GFAP. The results suggest that the age after which the lesioned area proves to be impenetrable for axons can precede that age after which lesions provoke glial reaction. In this case the inhibition of axon growth is to be attributed to factors other than to the reactive glia. The presence of nestin-positive cells suggests that the lack of reactive glia along the lesion track was not due to the absence of astrocytes, but rather due to the lack of their reaction to lesion. In this developmental stage astroglia, when activated, seem to promote the growth of axons. Accepted: 19 May 2000     

GFAP-immunopositive structures in spiny dogfish, Squalus acanthias, and little skate, Raia erinacea, brains: differences have evolutionary implications

GFAP expression patterns were compared between the brains of a spiny dogfish (Squalus acanthias) and a little skate (Raia erinacea). After anesthesia, the animals were perfused with paraformaldehyde. Serial vibratome sections were immunostained against GFAP using the avidin-biotin method. Spiny dogfish brain contained mainly uniformly-distributed, radially arranged ependymoglia. From GFAP distribution, the layered organization in both the telencephalon and the tectum were visible. In the cerebellum, the molecular and granular layers displayed conspicuously different glial structures; in the former a Bergmann glia-like population was found. No true astrocytes (i.e., stellate-shaped cells) were found. Radial glial endfeet lined all meningeal surfaces. Radial fibers also seemed to form endfeet and en passant contacts on the vessels. Plexuses of fine perivascular glial fibers also contributed to the perivascular glia. Compared with spiny dogfish brain, GFAP expression in the little skate brain was confined. Radial glia were limited to a few areas, e.g., segments of the ventricular surface of the telencephalon, and the midline of the diencephalon and mesencephalon. Scarce astrocytes occurred in every brain part, but only the optic chiasm, and the junction of the tegmentum and optic tectum contained large numbers of astrocytes. Astrocytes formed the meningeal glia limitans and the perivascular glia. No GFAP-immunopositive Bergmann glia-like structure was found. Astrocytes seen in the little skate were clearly different from the mammalian and avian ones; they had a different process system - extra large forms were frequently seen, and the meningeal and perivascular cells were spread along the surface instead of forming endfeet by processes. The differences between Squalus and Raia astroglia were much like those found between reptiles versus mammals and birds. It suggests independent and parallel glial evolutionary processes in amniotes and chondrichthyans, seemingly correlated with the thickening of the brain wall, and the growing complexity of the brain. There is no strict correlation, however, between the replacement of radial ependymoglia with astrocytes, and the local thickness of the brain wall.     

Evidence for the presence of the sphingosine-1-phosphate receptor Edg-8 in human radial glial fibers

The distribution pattern of Edg-8 immunostaining in the human developing brain has been investigated with special reference to radial glial fibers. At 24 weeks of gestation, fragments of radial glial fibers are Edg-8-positive within the cortical plate and subplate of allocortical areas. These Edg-8-positive fragments often appear enlarged as varicosities and some of them terminate at blood vessels. Between 28 and 30 weeks of gestation, all iso- and allocortical areas contain Edg-8-immunolabelled radial glial fibers revealing curvature next to sulci. After 32 weeks of gestation, radial glial fibers gradually disappear; instead Edg-8-positive transitional stages between radial glia and astrocytes were found. The findings indicate that sphingosine-1-phosphate may play a regulatory role in the transformation of radial glial cells into astrocytes and may affect proliferative activity of these cells.     

GFAP-immunopositive structures in spiny dogfish, Squalus acanthias, and little skate, Raia erinacea, brains: differences have evolutionary implications

GFAP expression patterns were compared between the brains of a spiny dogfish (Squalus acanthias) and a little skate (Raia erinacea). After anesthesia, the animals were perfused with paraformaldehyde. Serial vibratome sections were immunostained against GFAP using the avidin-biotin method. Spiny dogfish brain contained mainly uniformly-distributed, radially arranged ependymoglia. From GFAP distribution, the layered organization in both the telencephalon and the tectum were visible. In the cerebellum, the molecular and granular layers displayed conspicuously different glial structures; in the former a Bergmann glia-like population was found. No true astrocytes (i.e., stellate-shaped cells) were found. Radial glial endfeet lined all meningeal surfaces. Radial fibers also seemed to form endfeet and en passant contacts on the vessels. Plexuses of fine perivascular glial fibers also contributed to the perivascular glia. Compared with spiny dogfish brain, GFAP expression in the little skate brain was confined. Radial glia were limited to a few areas, e.g., segments of the ventricular surface of the telencephalon, and the midline of the diencephalon and mesencephalon. Scarce astrocytes occurred in every brain part, but only the optic chiasm, and the junction of the tegmentum and optic tectum contained large numbers of astrocytes. Astrocytes formed the meningeal glia limitans and the perivascular glia. No GFAP-immunopositive Bergmann glia-like structure was found. Astrocytes seen in the little skate were clearly different from the mammalian and avian ones; they had a different process system – extra large forms were frequently seen, and the meningeal and perivascular cells were spread along the surface instead of forming endfeet by processes. The differences between Squalus and Raia astroglia were much like those found between reptiles versus mammals and birds. It suggests independent and parallel glial evolutionary processes in amniotes and chondrichthyans, seemingly correlated with the thickening of the brain wall, and the growing complexity of the brain. There is no strict correlation, however, between the replacement of radial ependymoglia with astrocytes, and the local thickness of the brain wall. Accepted: 6 March 2001     

胰岛素对大脑皮质损伤后胶质细胞和AchE纤维变化的影响

本实验采用SD大鼠56只,于顶皮质作一宽2mm的冠状切口,用尼氏染色和AchE阳性纤维染色,观察损伤后胰岛素对切口两侧胶质细胞变化的影响.结果:损伤组和损伤用药组损伤切口两侧的胶质细胞在1～2W内迅速增生和集聚;2～3W出现胶质瘢痕,4～8W内增生的胶质细胞减少,胶质瘢痕收缩成一条致密带,再生胆碱能纤维沿切口两侧生长,难以穿越瘢痕组织,胰岛素用药组皮质损伤切口两侧的胶质细胞增生虽较损伤组轻,但还不足以有效的抑制胶质瘢痕的形成.     

Glial changes following an excitotoxic lesion in the CNS--II. Astrocytes.

Astrocytes are involved, as are microglia/macrophages [Marty et al. (1991) Neuroscience 45, 529-539], in the formation of a glial scar after CNS lesions. This study was undertaken to follow the time-course of changes in the morphology and distribution of astrocytes that takes place during the formation of a glial scar after kainic acid injection in the rat thalamus. The astrocytes were identified using an antibody raised against glial fibrillary acidic protein (GFAP) and the progression of their reaction to the lesion was followed from 24 h to one year after the kainate injection. Three periods could be distinguished during the evolution of the astrocytic response in the neuron-depleted area. There was an initial appearance of a large number of GFAP+ cells. These cells displayed profound morphological differences from the normal. They were enlarged, round and devoid of processes. These GFAP+ astrocytes disappeared four days after the lesion. This increase of the GFAP+ cells in the neuron-depleted area may be due to cytoskeletal changes and thus an increased exposure of antigenic sites. In a second period between four and 14 days, the only GFAP+ elements present in the neuron-depleted area were long and straight processes. These processes entered the lesioned area from the periphery and seemed to follow axon bundles. Additionally, during the first weeks, the number of reactive astrocytes increased in a small band just around the area of neuronal loss. The third period began after two weeks. The lesioned area became gradually occupied by GFAP+ astrocytes.(ABSTRACT TRUNCATED AT 250 WORDS)     

损毁单侧黑质-纹状体通路的大鼠SVZ、纹状体和黑质神经前体细胞的变化

研究成年大鼠脑室下区 (SVZ)神经前体细胞 (neural precursors)在黑质 -纹状体通路损伤后的反应 ,本研究用 6-羟多巴胺单侧纹状体注射以损毁黑质 -纹状体通路 ,损毁 10 d后腹腔注射 Brd U ,连续 4d,每日两次 ;在 SVZ、纹状体和黑质部位用免疫组化方法检测 Brd U、nestin以及 GFAP阳性细胞。结果显示 :(1) 6-羟多巴胺损毁黑质 -纹状体通路后 ,伤侧 SVZ的 Brd U阳性细胞数明显增多 ,并成簇分布 ;nestin和 GFAP阳性细胞数也增多 ,但以 GF AP阳性细胞增多明显 ;(2 )伤侧纹状体可见大量 Br-d U、GFAP以及少量 nestin阳性细胞分布 ,而健侧只有少量 GFAP阳性细胞 ;(3 )伤侧可见 Brd U阳性细胞在 SVZ和纹状体之间呈条带样分布 ;(4 )伤侧黑质除酪氨酸羟化酶阳性神经元减少外 ,未见 Brd U、GFAP和 nestin阳性细胞表达。上述结果表明 ,6-羟多巴胺损毁黑质 -纹状体通路后 ,SVZ神经前体细胞活动增强 ,有向纹状体迁移的趋势。     

Alterations in striatal glial fibrillary acidic protein expression in response to 6-hydroxydopamine-induced denervation

Following injection of 6-hydroxydopamine (6OHDA) into one side of the substantia nigra, immunohistochemical studies showed that the number of glial fibrillary acidic protein-positive [GFAP(+)] astrocytes in the striatum was significantly increased 1 day later and reached a maximum value, with intense immunoreactivity, 4 days after 6-OHDA injection. The number of GFAP(+) cells then gradually declined but was still 1.7 times the control value by 28 days postlesion. GFAP content, determined by immunoblot, and GFAP messenger RNA (mRNA) both reached maximal increases in the striatum 7 days after lesion: the mRNA returned to control values by 28 days, whereas GFAP content remained significantly elevated. Although the increases were all larger on the lesioned side, there were also significant changes on the contralateral side, as well as following saline injection. These results support the hypothesis that products released from damaged neurons are responsible for the induction of reactive gliosis, but cannot distinguish between effects mediated directly on the astrocytes or indirectly via other cells such as the microglia.     

An immunohistochemical study of the topography and cellular localization of three neural proteins in the sheep nervous system.

The peroxidase-anti-peroxidase (PAP) method was used to determine the topography and cellular localization of glial fibrillary acidic protein (GFAP), myelin basic protein (MBP) and carbonic anhydrase II (CAII) in the central nervous system (CNS), dorsal root ganglia and dorsal and ventral spinal nerve roots of the sheep. Parallel studies of mouse brain provided comparative data. Several fixatives were compared for their relative merits in preserving marker protein expression: GFAP was well preserved irrespective of the fixative employed; MBP was best preserved in formal sublimate and CAII was best preserved in Carnoy's fluid. In sheep, GFAP expression was seen in protoplasmic and fibrous astrocytes, Bergmann glial cells, a proportion of ependymal cells, amphicytes of spinal ganglia and in a proportion of presumed Schwann cells of dorsal and ventral spinal nerve roots. MBP expression was seen in mature and developing myelin sheaths of the central nervous system and in the cytoplasm of sparse myelinating oligodendroglia of the sub-cortical white matter of the cerebrum. CAII expression was seen in choroid plexus epithelium in all ages of sheep studied and, in a young lamb and an adult sheep, in glia and neuropil of ventral horn grey matter of the spinal cord and in the cytoplasm of white matter glia, presumed fibrous astrocytes, throughout the CNS. Compared with sheep brain, mouse brain showed the following differences in marker protein localization. GFAP was weakly expressed by protoplasmic astrocytes and not expressed in ependyma, oligodendroglia expressing intracytoplasmic MBP were frequent and widespread in neonatal mouse brain, CAII was expressed in myelin and oligodendroglia.(ABSTRACT TRUNCATED AT 250 WORDS)