Expression of glial fibrillary acidic protein (GFAP) by cultured angiofibroma stroma cells from patients with tuberous sclerosis

Large dendritic cells were cultured from facial angiofibromas of six patients with tuberous sclerosis. The cells were examined immunocytochemically for expression of selected cytoskeletal and non-structural proteins and the results compared with the staining profiles obtained with normal skin fibroblasts and normal glial cells. In similarity to normal glia, the angiofibroma stroma cells expressed glial fibrillary acidic protein (GFAP). Conversely, by analogy to fibroblasts, the abnormal stroma cells produced fibronectin and did not react with the antibody to S-100 protein. By immunogold labelling it was established that GFAP and vimentin were co-localized in intermediate filaments of the angiofibroma cells.     

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.     

Insulin influences astroglial morphology and glial fibrillary acidic protein (GFAP) expression in organotypic cultures.

Variations in the levels and timing of exposure to insulin-related peptides influence the phenotypic appearance of astroglia present in organotypic cultures of the E17 mouse cerebellum as well as the expression of glial fibrillary acidic protein (GFAP) mRNA and its encoded protein. The morphology of GFAP-immunoreactive cells was influenced by the levels of insulin added in an age-specific manner. Fetal radial glia were selectively and significantly (P less than 0.001) increased by high (10 micrograms/ml) insulin levels, comprising the majority of the GFAP-positive cells seen. In contrast, there was an almost complete reversal of this pattern elicited by low (10 pg/ml) insulin levels, where GFAP-positive cells appeared undifferentiated and epithelioid (P less than 0.001). In newborn cultures, on the other hand, the morphological responses to both high and low levels of insulin were considerably attenuated and involved radial glia primarily, whose numbers were significantly increased by the high insulin levels. Exposure to high levels of insulin was accompanied by an increase in GFAP mRNA expression, as determined by non-isotopic (biotin) in situ hybridization histochemistry, and intense GFAP immunoreactivity, while low insulin levels elicited minimal expression of both message and protein product. In view of the critical interdependence of developing neurons and radial glia with respect to neuronal migration and the differentiation of neurons and astroglia, the responses observed suggest developmentally regulated mechanisms by which insulin-related peptides themselves may influence directly and indirectly both neuronal and astroglial differentiation.     

Expression of glial fibrillary acidic protein and its relation to tract formation in embryonic zebrafish (Danio rerio)

To address possible roles of glial cells during axon outgrowth in the vertebrate central nervous system, we investigated the appearance and distribution of the glial-specific intermediate filament, glial fibrillary acidic protein (GFAP), during early embryogenesis of the zebrafish (Danio rerio). Immunopositive cells first appear at 15 hours, which is at the time of, or slightly before, the first axon outgrowth in the brain. Immunopositive processes are not initially present in a pattern that prefigures the location of the first tracts but rather are distributed widely as endfeet adjacent to the pia, overlying most of the surface of the brain with the exception of the dorsal and ventral midline. The first evidence for a specific association of immunopositive cells with the developing tracts is observed at 24 hours in the hindbrain, where immunopositive processes border axons in the medial longitudinal fasciculus. By 48 hours, immunopositive processes have disappeared from most of the subpial lamina and are found exclusively in association with tracts and commissures in three forms: endfeet, radially oriented processes, and tangentially oriented processes parallel to axons. This last form is particularly prominent in the transverse plane of the hindbrain, where they define the boundaries between rhombomeres. These results suggest that glial cells contribute to the development and organization of the central nervous system by supporting early axon outgrowth in the subpial lamina and by forming boundaries around tracts and between neuromeres. The results are discussed in relation to previous results A neuron-glia interactions and possible roles of glial cells in axonal guidance. © 1995 Wiley-Liss, Inc.     

Involvement of glial fibrillary acidic protein (GFAP) expressed in astroglial cells in circadian rhythm under constant lighting conditions in mice

To clarify the role of glial fibrillary acidic protein (GFAP)-expressed glial cells in the circadian clock, we examined GFAP expression in the suprachiasmatic nucleus (SCN) and the intergeniculate leaflet (IGL) under various lighting conditions in mice. We demonstrated that GFAP expression did not show daily change in the SCN under a light-dark cycle; however, long-term housing under constant lighting conditions led to dramatic changes in GFAP expression, i.e., a decrease in the SCN and an increase in the IGL. Furthermore, mice that had a targeted deletion in the GFAP gene (GFAP mutant mice) showed longer and more arrhythmic circadian activity rhythms in constant lighting conditions than wild-type mice, while GFAP mutant mice exhibited stable circadian rhythms both in a light-dark cycle and constant darkness, and showed normal entrainment to environmental light stimuli. These results suggest that the GFAP-expressed astroglial cells in the SCN and the IGL may have some role in circadian oscillation under constant lighting conditions.     

Patients with suspected benign tumors and glial fibrillary acidic protein autoantibody: an analysis of five cases

Abstract

Aim: To investigate the clinical features of five glial fibrillary acidic protein (GFAP) antibody positive patients with suspected benign tumors and explore its underlying pathogenesis.

Materials and methods: Overall, 1018 serum and cerebrospinal fluid (CSF) samples were tested by indirect immunofluorescence assay and data from five patients with suspected tumors and positive for GFAP autoantibody in the CSF were analyzed retrospectively.

Results: The positive rate of GFAP antibody in the serum and CSF was 3.93% by indirect immunofluorescence assay. Tumors were diagnosed before or after neurologic onset in 5 of 40 patients (12.5%) and no deterioration of the tumors was found during the long-term follow-up. Of the five patients, one patient suffered a thyroid nodule, one patient had a small nodule in the left lung, two patients suffered meningiomas, and one patient had a suspicious eosinophilic granuloma.

Conclusion: GFAP autoimmunity may be a paraneoplastic immune response with a low frequency of tumor in Chinese patients with GFAP astrocytopathy.     

Methylprednisolone inhibits the expression of glial fibrillary acidic protein and chondroitin sulfate proteoglycans in reactivated astrocytes

Reactive gliosis caused by post-traumatic injury often results in marked expression of chondroitin sulfate proteoglycan (CSPG), which inhibits neurite outgrowth and regeneration. Methylprednisolone (MP), a synthetic glucocorticoid, has been shown to have neuroprotective and anti-inflammatory effects for the treatment of acute spinal cord injury (SCI). However, the effect of MP on CSPG expression in reactive glial cells remains unclear. In our study, we induced astrocyte reactivation using alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and cyclothiazide to mimic the excitotoxic stimuli of SCI. The expression of glial fibrillary acidic protein (GFAP), a marker of astrocyte reactivation, and CSPG neurocan and phosphacan were significantly elevated by AMPA treatment. The conditioned media from AMPA-treated astrocytes strongly inhibited neurite outgrowth of rat dorsal root ganglion neurons, and this effect was reversed by pretreatment with MP. Furthermore, MP downregulated GFAP and CSPG expression in adult rats with SCI. Additionally, both the glucocorticoid receptor (GR) antagonist RU486 and GR siRNA reversed the inhibitory effects of MP on GFAP and neurocan expression. Taken together, these results suggest that MP may improve neuronal repair and promote neurite outgrowth after excitotoxic insult via GR-mediated downregulation of astrocyte reactivation and inhibition of CSPG expression.     

Proliferation of gemistocytic cells and glial fibrillary acidic protein (GFAP)-positive oligodendroglial cells in gliomas: a MIB-1/GFAP double labeling study

Large gemistocytic cells are well-known elements of glial tumors. Recently, miniature gemistocytic cells and neoplastic glial fibrillary acidic protein (GFAP)-positive oligodendroglial cells, which are regularly seen in oligodendrogliomas, have been termed "transitional cells". The proliferative activity of the gemistocytic cell types and the GFAP-positive (gliofibrillary) oligodendrocytes was determined in eight astrocytomas, seven gemistocytic astrocytomas, eight glioblastomas, two monstrocellular glioblastomas, seven oligodendrogliomas and three mixed oligo-astrocytomas by immunohistochemical staining of the proliferation marker MIB-1 in combination with immunostaining for GFAP. Both large gemistocytic cells and the transitional cells showed cytoplasmic GFAP-positive staining. Neither in the classic gemistocytes nor in the minigemistocytes nuclear immunostaining for the MIB-1 antibody was observed. In contrast, MIB-1 staining was seen in the gliofibrillary oligodendrocytes. It is concluded that both large and miniature gemistocytic cell types contrast with gliofibrillary oligodendrocytes by their inability to proliferate. Received: 16 March 1995 / Revised: 24 May 1995 / Revised, accepted: 21 August 1995     

PKA and PKC activation induces opposite glial fibrillary acidic protein (GFAP) expression and morphology changes in a glioblastoma multiform cell line of clonal origin

Possible differentiation mechanisms were investigated in a glioblastoma multiform cell line (GL15) presenting an undifferentiated phenotype with weak glial fibrillary acidic protein (GFAP) and strong vimentin (VIM) expression. Serum-free conditions induced time-dependent increases of GFAP-mRNA and GFAP protein levels, associated with a process-bearing astrocytic morphology. Activation of protein kinase C (PKC) by tumor promoter phorbol 12-myrystate 13-acetate (PMA) induced a rapid morphological differentiation and a decrease in GFAP mRNA, whereas the GFAP level remained unchanged. Such parameters were shown to characterize a physiological differentiation stage in astroglial cultures. Treatment of process-bearing GL15 cells with dibutyryl cyclic AMP (dbcAMP), a protein kinase A (PKA) activator, induced a timedependent decrease in the GFAP mRNA and GFAP protein levels and reverted morphological changes induced by serum-free conditions. Neither PMA nor dbcAMP influenced the VIM mRNA expression. In GL15 cells, PKC and PKA activation have opposite effects. Understanding the role of these kinases in malignant transformation and in the in vitro differentiation process is of both basic and clinical interest. © 1995 Wiley-Liss, Inc.     

High cerebrospinal fluid concentration of glial fibrillary acidic protein (GFAP) in patients with normal pressure hydrocephalus

The concentration of glial fibrillary acidic protein (GFAP) in lumbar cerebrospinal fluid (CSF) was measured in 12 patients with normal pressure hydrocephalus (NPH) 11 patients with primary degenerative dementia (PDD), 8 patients with various other neurological diseases, and 18 patients without signs of organic nervous disease (controls).

Mean CSF GFAP concentration was significantly higher in NPH patients: 96 ± 23 ng/ml (SEM) when compared with PDD patients: 8.2 ± 1.9 ng/ml (P < 0.01), or with controls: 4.3 ± 0.7 ng/ml (P < 0.01). Only 2 NPH patients had a GFAP concentration within the range of the control group (2–14 ng GFAP/ml CSF). No significant differences were found between the PDD patients and the control group, or between the group of patients with other neurological diseases and the control group.

In addition, a rostro-caudal gradient of GFAP in CSF could be demonstrated. In 6 NPH and 2 PDD patients both ventricular and lumbar CSF samples were investigated. In all cases the ventricular GFAP concentration was higher than the lumbar concentration. The difference was statistically significant (P < 0.01).

Our results suggest that determination of CSF GFAP concentration might be of diagnostic value in discrimination between NPH patients and patients with enlarged ventricles associated with degenerative brain disease.     

正常大鼠脑组织伽玛刀照射后胶质细胞原纤维酸性蛋白的表达及变化

目的:观察正常大鼠脑受伽玛刀(γ-刀)照射后星形胶质细胞的反应。方法:45只正常成年大鼠脑接受γ-刀100Gy量的照射,分别成活0.5h、1h、3h、6h、12h、24h、3d、7d、14d、30d和3个月后被处死,固定取脑,冰冻切片,进行抗胶质细胞原纤维酸性蛋白(GFAP)的免疫组织化学反应。结果:①γ-刀照射后在靶区和海马等处的GFAP免疫反应(GFAPir)阳性细胞增多;②可见到2种阳性细胞,一种是胞体小、突起细短的细小型,另一种是胞体肥大、突起粗长、染色深的肥大型,存活时间越长,越靠近靶中央区,肥大型GFAPir细胞越多,肿胀越明显,反之则少;③GFAPir表达有2次高峰,第一次在照射后24h以内,第二次在照射后14d以后;④从照射后14d起海马内GFAPir细胞有的部位脱失,形成“斑秃”样分布。结论:以上结果表明GFAPir细胞在脑内的数量、形态及分布特点可以作为脑损伤的一种标志。     

Spinal cord multiple sclerosis lesions in Japanese patients: Schwann cell remyelination occurs in areas that lack glial fibrillary acidic protein (GFAP)

Summary To extend earlier observations on Schwann cell remyelination in multiple sclerosis (MS) lesions (Itoyama et al. 1983) we immunostained spinal cord sections from eight Japanese MS patients with antiserum to P_o glycoprotein, a major constituent of peripheral nervous system (PNS) myelin, myelin basic protein (MBP), and glial fibrillary acidic protein (GFAP). Spinal cord sections from six of the eight Japanese MS patients contained large clusters of peripheral myelin sheaths with anti-P_o immunoreactivity. In lesions found in four of the six patients, thousands of P_o-stained PNS myelin sheaths were present. Necrosis was prominent in these lesions which included more than half of the spinal cord's transverse area. The number and density of regenerating myelin sheaths of peripheral origin were much greater than we observed in MS spinal cord lesions of white people (Itoyama et al. 1983). Anti-GFAP immunoreactivity was present in most brain and spinal cord lesions. However, the areas in lesions that contained large groups of PNS myelin sheaths lacked anti-GFAP immunoreactivity. Our data suggest that spinal MS lesions that are large, severely demyelinated, and partially necrotic may contain factors that inhibit fibrous astrogliosis. These factors, other substances in the large lesions and/or the lack of astrocytic scarring could then promote Schwann cell invasion, multiplication, and remyelination of surviving axons.     

Distribution of glial fibrillary acidic protein (GFAP) in the cochlear nucleus of adult and aged rats

The age-related change in glial fibrillary acidic protein (GFAP) immunoreactivity was analyzed in young (3 months) and old (24 months) adult rat cochlear nuclei (CN). Quantitative analyses show a significant increase with age, in the number of GFAP positive astrocytes and processes in the old adult when compared with the young adult rat. There was also a differential distribution of GFAP immunoreactivity in the young adult CN where it predominates in the granular cell region, whereas in old rats, the GFAP immunoreactivity distribution was homogeneous in all parts of the nucleus. There was no change in the total number of neurons between these two stages in any part of the nucleus except for the antero-ventral CN, where a decrease in neuronal number was observed in the aged rats. The increase in GFAP immunoreactivity was related to an increase of both GFAP positive astrocyte number and processes. The increase of GFAP positive astrocytes may be due either to an alteration of auditory nerve fibers, changing the trophic interactions with post-synaptic cells, or to intrinsic alterations of CN neurons and local circuits reflecting aging of the CN.     

Late-onset Alexander disease with a V87L mutation in glial fibrillary acidic protein (GFAP) and calcifying lesions in the sub-cortex and cortex

Glial fibrillary acidic protein (GFAP) mutation has been reported in Alexander disease. We report a 31-year-old woman suffering from Alexander disease with a V87L mutation in GFAP. She showed psychomotor regression and a history of seizures, in addition to pendular nystagmus, dysarthria, spastic gait, and bladder dysfunction. Brain magnetic resonance imaging (MRI) showed atrophy of the medulla oblongata and mild cervical cord atrophy, deep white matter abnormalities, periventricular rim, and signal changes of the medulla oblongata and dentate hilum. Sequence analysis of her GFAP gene showed a heterozygous c.273G>C mutation predictive of a p.V87L amino acid substitution. We concluded that she was actually affected with Alexander disease. Twenty months later she fell down and sustained a head contusion. Urgent head computed tomography (CT) showed calcification in the subcortical and cortical regions, which may relate to the psychomotor regression and history of seizures. Calcification in the subcortical and cortical regions on head CT has not been reported in Alexander disease; this may be associated with a V87L mutation in GFAP.     

Immunocytochemical localization of glial fibrillary acidic protein (GFAP) in the area postrema of the cat. Light and electron microscopic study

Glial fibrillary acidic protein (GFAP) was demonstrated in the cytoplasm and processes of ependymal cells and astroglial components of the area postrema of the cat. These observations differ from the findings in the ependyma of the ventricular cavities which are consistently negative for the protein. Since some studies have suggested sensory functions of the glial cells in this emetic chemoreceptor trigger zone, a careful consideration of morphological and biochemical attributes of these cells seems appropriate.     

Evidence that formation of an intermediate filament-protein complex plays a primary role in aggregation of neurofilaments, glial fibrillary acidic protein (GFAP)-filaments and vimentin-filaments by 2,5-hexanedione

Using a variety of cell types in culture, we have investigated the relevance of intermolecular crosslinking involving intermediate filament proteins (IF-proteins) to the changes in distribution of intermediate filaments induced by the neurotoxicant, 2,5-hexanedione (2,5HD). Aggregation of vimentin-filaments (vimentin-IF), glial fibrillary acidic protein (GFAP)-IF and neurofilaments was preceded by appearance on immunoblots of a high molecular weight complex (IF-complex) labeled by antibodies against the IF-protein of the cell type: pV-170 from human skin fibroblasts and pV-130 from 3T3 mouse fibroblasts, which were labeled by anti-vimentin and, from cultures of dissociated mouse spinal cord and dorsal root ganglia, pNFH-300 labeled by antibody against the 200 kDa neurofilament protein (NF-200 or NF-H) and a doublet labeled by antiserum to GFAP (pGFAP-145).pV-170 was detected in human skin fibroblasts within one hour of exposure to 2,5HD and the amount of both pV-170 and pNFH-300 was related to the concentration of 2,5HD and the duration of exposure. Intermediate filament-complexes were not detected in PtK1 epithelial cells by labeling of transblots with anti-cytokeratin, nor are keratin-IF aggregated by 2,5HD. Intermediate filament-complexes were not detected in fibroblasts with IF-aggregates secondary to disruption of microtubules by colchicine or in fibroblasts from patients with giant axonal neuropathy.