Retardation in somatosensory cortex development induced by postnatal BrdU treatment in mice

Cerebral dysgeneses are in the background of several neurological and mental disturbances. The aim of the present study was to investigate structural and activity changes following disturbed postnatal neuronal development in mice. Newborn C57Bl6 mice were exposed to 5-bromo-2′-deoxyuridine (BrdU: daily 50 μg/g body weight) during a period between postnatal days P0–P5 or P0–P11, respectively, and neuronal malformation and malfunctioning of somatosensory (barrel field) cortex was analyzed in adolescent animals. Alterations in histological architecture of interneuronal and glial elements were studied and correlated with electrophysiological modifications. Between P30 and P35 days litters underwent ex vivo electrophysiological experiments to examine the changes in basic excitability and in synaptic efficacy. Parallel immunohistochemistry was performed to detect BrdU, GABA and GFAP.     

Review of Alexander disease: Beyond the classical concept of leukodystrophy

Alexander disease is classified as one of the leukodystrophies, which are degenerative diseases primarily affecting the cerebral white matter. Formal diagnosis is achieved by showing diffuse accumulation of Rosenthal fibers in the brain by biopsy or autopsy. Showing a heterozygous mutation in the glial fibrillary acidic protein (GFAP) gene is currently sufficient for diagnosis. The mechanisms of Rosenthal fiber formation remain unclear. However, both the quality and quantity of GFAP are important. GFAP-ε (rodent homologous GFAP-δ), one of the alternatively spliced GFAP isoforms, may also play a modulating role in aggregate formation. The current ease of diagnosis has accelerated the accumulation of a wide variety of patients with Alexander disease along with the widespread use of MRI. In contrast to the classical infantile type, patients with juvenile and adult types mainly complain of bulbar symptoms and usually show progressive atrophy of the lower brainstem and cervical spinal cord with mild or minimal leukodystrophic changes. Among the many MRI findings of Alexander disease, periventricular linear lesions with various names depending on the thickness and shape seem to represent the unique pathophysiology, because the subventricular zone of the adult human brain includes special astrocytes that behave as multipotent progenitor cells and specifically produce GFAP-ε. Except for a few mutations, no clear phenotype–genotype correlation has been established for Alexander disease, although male preponderance in the infantile type suggests that phenotypes may be partly affected by gender.     

Growth activity of tumors at different intracranial structures: immunohistochemical study with bromodeoxyuridine

Summary To elucidate the environmental influence on the growth of a tumor, bromodeoxyuridine (BrdU) uptake in multiple tumor foci within the intracranial cavity was studied immunohistochemically with a monoclonal antibody. Walker 256 tumor implanted intracerebrally produced multifocal tumors presented as (1) intraparenchymal solid tumor, (2) tumor in the choroid plexus, and (3) leptomeningeal dissemination. The BrdU-labeling indices, or the S-phase fractions (% of nuclei labeled by BrdU divided by the number of tumor cell nuclei scored; LI), of those tumors were 48.4±1.1, 59.1±1.3, and 27.9±5.9, respectively (means ± SEM). These differences in LI, or the tumor growth activity, are discussed in relation to the different environmental conditions in different host structures. These host structure-related modification of tumor growth would be important in evaluating the proliferative activity of tumors growing at various intracranial structures.Supported in part by the Grant-in-Aid for Cancer Research (59-22) from Ministry of Welfare     

Cerebromeningeal pleomorphic xanthoastrocytoma. Report on four cases: Clinical,radiologic and pathological features. (Including a case with malignant evolution)

Three cases of cerebromeningeal pleomorphic xanthoastrocytoma are presented and the recurrence of a fourth case, previously described in 1980, is examined because of its evolution towards malignancy.Clinical, radiologic and pathological aspects are discussed and the general features of the previously reported cases of xanthoastrocytoma are reviewed.     

Response of rat cerebral cortical astrocytes to freeze- or cobalt-induced injury: an immunocytochemical and gap-FRAP study

Astrocytic response in the immediate vicinity of freeze- and cobalt-induced lesions has been examined at the light and ultrastructural level. However, the temporal and spatial distribution of astrocytic reactivity throughout the rat cerebral cortex, using glial fibrillary acidic protein (GFAP) immunolabeling, has not been examined. The first purpose of this study was to establish the chronological distribution of astrocytic reactivity, as measured by changes in GFAP immunoreactivity, following freeze- or cobalt-induced injury to the rat cerebral cortex. Cobalt metal also has been proposed to have a direct effect on astrocytes and has been shown to stimulate in vitro astrocytes to become reactive. The second purpose of this report was to determine if cobalt had an effect on in vitro astrocytic gap junctional dye coupling as measured by fluorescence recovery after laser-photobleaching (gap-FRAP). Although the chronological development of the increased GFAP immunoreactivity was different for the freeze- and cobalt-induced lesions, astrocytes initially showed an increase in GFAP immunoreactivity in the region surrounding these lesions. This initial response was followed by a spread of increased GFAP immunoreactivity throughout certain regions of the ipsilateral cerebral hemisphere and then by a restriction of the increased immunolabeling to the lesion site. Cobalt also had a direct effect on in vitro astrocytes as demonstrated by the inhibition of astrocytic gap junctional dye coupling. Based on gap-FRAP analysis, cobalt significantly blocked fluorescence recovery (2.5%) as compared to the fluorescence recovery in control astrocytes (26%). It is proposed that the initial increase in GFAP immunoreactivity may be due to decreased gap junctional activity.     

Neuroinflammation and astrocytic reaction in the course of Phoneutria nigriventer (armed-spider) blood–brain barrier (BBB) opening

Phoneutria nigriventer spider venom (PNV) causes uneven BBB permeability throughout different cerebral regions. Little is known about cellular and molecular responses which course with the PNV-induced BBB opening. We investigate by immunohistochemistry (IHC) and Western blotting (WB), the GFAP, S100, IFN-γ and TNF-α proteins expression in hippocampus and cerebellum after different time-points from venom or saline intravenous injection. All proteins variably altered its expression temporally and regionally. WB showed increased GFAP content at 15–45 min followed by a shift below the control level which was less pronounced in hippocampus. IHC showed reactive gliosis during all the trial period. In cerebellum, GFAP was mostly immunodetected in astrocytes of the molecular layer (Bergmann glia), as was S100 protein. The maximum S100 immunolabeling was achieved at 5 h. IFN-γ and TNF-α, expressed mostly by hippocampal neurons, increased along the trial period, suggesting a role in BBB permeability. In envenomed animals, closer contacts astrocyte–astrocyte, granule cells–granule cells and astrocytes-Purkinje cells were observed in cerebellum. Closer contacts between neurons–neurons–astrocytes–astrocytes were also seen in hippocampus. PNV contains serotonin, histamine, Ca²⁺ channels-blocking toxins, some of which affect glutamate release. The hypothesis that such substances plus the cytokines generated, could have a role in BBB permeability, and that calcium homeostasis loss and disturbance of glutamate release are associated with the marked GFAP/S100 reaction in Bergmann glia is discussed. The existence of a CNS mechanism of defense modulated differentially for fast synthesis and turnover of GFAP, S100, IFN-γ and TNF-α proteins was evident. A clear explanation for this differential modulation is unclear, but likely result from regional differences in astrocytic/neuronal populations, BBB tightness, and/or extent/distribution of microvasculature and/or ion channels density/distribution. Such differences would respond for transient characteristics of BBB disruption. This in vivo model is useful for studies on drug delivery throughout the CNS and experimental manipulation of the BBB.     

Bombycis corpus Extract (BCE) Protects Hippocampal Neurons Against Excitatory Amino Acid-Induced Neurotoxicity

Bombycis corpus (BC) or Bombyx Batryticatus, a batryticated silkworm and white-stiff silkworm, is a drug consisting of the dried larva of silkworm, Mobyz mori L., dead and stiffened due to the infection of Beauveria (Bals.) Vuill. In a previous paper (Kim et al., Pharmacol. Res., 43, 12-16, 2001), BC was shown to protect amyloid-β-induced cytotoxicity. In the present study, we have found that BCE can prevent or reduce the neurotoxic actions in the hippocampus of the glutamate agonists N-methyl-D-aspartic acid (NMDA) in vitro or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainic acid in vitro. Pre-treatment with BCE (0, 1, 2, 5, and 10 μg/ml for 6-8 h) protected primary hippocampal cultures from embryonic day 18 (E18) embryos against NMDA-induced toxicity (0.1, 1, 10, and 50 mM/ml). BCE added either with NMDA (1 mM) or 1 h later had lesser, but still significant, protective actions. BCE also reduced NMDA-induced toxicity (1 mM). BCE (10 μg/ml) protected cultured neurons against the neurotoxic actions of either AMPA (25 μM) or kainic acid (1 mM) as well. Because the release of glutamate has been implicated in the neural damage after cerebral ischemia and other neural insults, these results suggest that BCE may contribute significantly to protect human brain to such damage.     

新生儿肠闭锁NT-3与Trk-C表达研究

目的了解神经节细胞NT-3、Trk-C、Nf蛋白和神经胶质细胞GFAP蛋白在肠闭锁中的分布情况,并探讨其临床意义.方法应用免疫组化技术检测15例先天性小肠闭锁患儿肠壁内NT-3、Trk-C、GFAP和Nf的表达情况,并以死于与肠道发育异常无关的尸检新生儿小肠标本6例作为正常对照.结果肠闭锁组肌间神经丛NT-3阳性细胞数明显低于对照组的阳性细胞数(t'=9.9000,P<0.05).Trk-C在对照组肠壁肌间神经丛中存在高表达,而在肠闭锁组存在低表达(t=22.2991,P<0.05).对照组肠壁肌间神经丛胶质细胞GFAP呈阳性表达,而肠闭锁组GFAP呈强阳性表达.Nf在对照组中的表达明显高于闭锁组.结论先天性小肠闭锁的近端肠壁内存在有神经节细胞的减少和神经干的变细,支持细胞在闭锁近端肠壁中呈增生表现.这种肠壁肌层神经丛的改变,可能会引起肠腔内压力减低,从而影响闭锁区域肠道的功能.     

Heterochronous maturation of regional brain astroglia: neuronal modulation of striatal glial cells differentiation ex vivo

Subcultured astroglial cells from striatum, cerebral cortex and ventral mesencephalon obtained from primary cultures of fetal (E14, E17 and E21) or postnatal (days 5-6) rats showed different regional, age-dependent morphological response (stellation) to cyclic AMP. While most of the cerebral cortex and ventral mesencephalic astroglial cell population was responsive at all ages tested, striatal cells at E14 and E17 were not. At age E21 striatal astroglia showed a significant shift toward a mature-like type of response to cyclic AMP. Postnatal striatal astroglia responded to cyclic AMP as the cortical and ventral mesencephalic astroglia did, with generalized stellation. Prenatal striatal astroglia was characterized immunocytochemically as A2B5+, fibronectin+, vimentin+, S-100+ and GFAP-. Failure of early prenatal (E14, E17) striatal astroglia to differentiate in response to cyclic AMP, was overcome by previous (5-7 days) co-culture with primary cell dissociates from postnatal-, but not from prenatal donors, from all brain regions tested including a non-target region for striatal cells, such as septum. This effect was duplicated when striatal astroglia was co-cultured with cell populations enriched in neurons through Percoll gradients. Only cell-to-cell contact co-cultures were able to induce a change in the studied response. Dead neuron-enriched populations obtained following various types of physical treatments were also able to change significantly striatal cell response toward cyclic AMP. Enriched astroglial populations from postnatal donors did not change striatal astroglial response toward cyclic AMP, except for ventral mesencephalic astroglia which induced a comparatively reduced but significant increase in striatal cell responsiveness. It is concluded that astroglial maturation and potential for phenotype expression during brain development proceeds with regional heterochrony. Also, that maturation of prenatal striatal astroglia responsiveness toward cyclic AMP is inducible by non-diffusible factors, probably of neuronal origin, expressed in live or dead primary cultures from various, homotopic and heterotopic, postnatal brain regions. It is further suggested that striatal afferents and/or mature local striatal neurons express membrane associated molecules that regulate responsiveness for phenotype expression of striatal glial cells, thus reinforcing the concept of a highly interactive, continuous neuron-glial developmental process that takes place during brain organization.     

BMP7蛋白对大鼠急性脊髓损伤后神经元修复作用的研究

目的探索骨形态发生蛋白7(bone morphogenetic protein 7,BMP7)是否具有促进大鼠急性脊髓损伤后神经元修复的作用。方法实验大鼠分为阴性对照组(negative control,NC)和BMP7实验组,以Allen's打击法建立大鼠脊髓损伤模型,BMP7组大鼠每天在脊髓损伤处注射50ng BMP7蛋白,连续7天,NC组对应注射等体积的0.9%氯化钠注射液。两组大鼠分别于术后6小时、3天、1周、2周、4周、8周进行HE染色以观察脊髓损伤处病理学改变;两组大鼠分别于术后6小时、3天、1周、2周、4周、8周进行免疫组化实验以检测损伤节段脊髓神经丝蛋白200(neurofilament protein 200,NF200)、胶质纤微酸性蛋白(glial fibrillary acidic protein,GFAP)及突触素(Synaptophysin,SYP)蛋白表达水平。结果 HE染色结果表明两组大鼠造模后脊髓损伤处神经元数量减少,神经突触数量减少,尼式体淡染且数量减少,组织中可见空洞形成,1周后,BMP7实验组大鼠脊髓损伤处尼式体染色逐渐加深,神经突触数量增多,变化较NC组明显。免疫组化结果表明BMP7实验组大鼠NF200阳性表达细胞数造模3天后逐渐增多,至第4周时达到高峰,且在1周、2周、4周及8周时均大于NC组;BMP7实验组及NC组术后GFAP阳性表达细胞数变化均不明显,且两组阳性细胞数差异不显著;BMP7实验组大鼠SYP阳性表达细胞数造模3天后逐渐增多,且在1周、2周、4周及8周时均大于NC组。结论 BMP7蛋白可以促进大鼠脊髓损伤后神经元的修复。