Glial Fibrillary Acidic Protein Activation Research Articles

A 1.8kb GFAP-promoter fragment is active in specific regions of the embryonic CNS.

The intermediate filament glial fibrillary acidic protein (GFAP) constitutes the major cytoskeletal protein in astrocytes (J. Neuroimmunol. 8 (1985) 203) and is traditionally referred to as a specific marker for astrocytes. To identify early glial precursors, we created GFAPpromoter-lacZ transgenic mice, using a 1.8kb 5' fragment of human GFAP. The expression of the transgene was first detected in the neuroepithelium at embryonic day 9.5. It was further found in the ventricular zone of the developing telencephalon, in the cerebellar primordium, trigeminal ganglia, and radial glia. Later, scattered beta-gal+ cells were seen in pons, brain stem and glia limitans. The results indicate that GFAP activity is regulated in a region-specific manner during central nervous system (CNS) development and that the gene is turned on in different cell types independently.

Delayed induction of JunB precedes CA1 neuronal death after global ischemia in the gerbil

The immediate early genes (IEGs), c- jun, junB and c- fos are expressed after global ischemia in the gerbil. The role of these genes remains unclear. Whilst moderate ischemia (7 min) causes delayed CA1 neuronal death, pre-conditioning with mild ischemia (2 min) neuroprotects the CA1 subfield. This differential response allows the specific expression patterns of IEGs to be associated with either delayed neuronal death, or cell survival, depending upon the insult severity. Using a graded insult strategy we have shown that (1) early IEG expression is prominent in the neuronal layers of the CA3, hilar and dentate regions, and (2) a delayed, secondary wave of JunB expression is localized to the selectively vulnerable CA1 neuronal layer after moderate ischemia. This expression precedes the histological and histochemical features of neuronal death. Delayed JunB expression was not observed in animals subject to 2 min ischemia. The glial fibrillary acidic protein (GFAP) promotor possesses an AP-1 binding site, the target for IEG dimers. To examine the possible link between IEG expression and astrocyte activation the transcriptional activation of GFAP was assessed. GFAP mRNA was evident within 8 h of ischemia after both insults. The extent of the astrocytic reaction was dependent upon the severity of the ischemia. The temporospatial distribution of IEG and GFAP expression differed, indicating that glial activation is unlikely to be regulated by the hippocampal expression of IEGs. We conclude that early IEG expression is involved in signalling mechanisms that invoke neuroprotective effects in the dentate and CA3 regions, and that delayed JunB expression in the CA1 subfield is associated with neuronal death, and may be involved in the commitment or execution phases of cell death. Early astrocytic responses may play a role in the mechanism of ischaemic tolerance.

Distribution of growth associated protein (B-50/GAP-43) and glial fibrillary acidic protein (GFAP) immunoreactivity in rat homotopic olfactory bulb transplants.

Recent studies on olfactory bulb homotopic transplantation after partial or subtotal bulb ablation have shown that regenerated olfactory axon are able to form glomeruli-like structures either in the transplant or in the spared olfactory bulb. To investigate the maturation of olfactory axons and their terminal connections in the transplant and remnant olfactory bulb we performed immunohistochemistry utilizing as markers B-50/GAP43 for neurite outgrowth and glial fibrillary acidic protein (GFAP) for the glial response to the surgery. Radioactively prelabeled olfactory bulbs (E 18) were homotopically transplanted in unilaterally bulbectomized neonatal rats (P6). Two to four months after transplantation in the partially bulbectomized rats the laminar organization of the olfactory bulb remnant depended on the extent of lesion. The transplant was disorganized showing irregularly distributed glomeruli. In a few cases pseudo-laminar organization was observed resembling that of the normal olfactory bulb. In 2 month-old transplants, outgrowing axons had a newly formed glomeruli displayed prominent B-50/GAP43 immunoreactivity. Four months after the operation B-50/GAP43 immunoreactivity was still present in the regrowing axons and some glomeruli in the transplant were B-50/GAP43 positive, while other glomeruli revealed a patchy pattern. The same B-50/GAP43 immunostained patchy structure were present in remnants of the lesioned olfactory bulb. Distinct increase of GFAP activity was observed along the olfactory axons and in the glomeruli-like structures. The persisting B-50/GAP43 immunoreactivity in the glomeruli of the transplant and in the remnants of lesioned olfactory bulb suggests that maturation of the newly formed glomeruli was delayed in comparison to the intact control olfactory bulb. Furthermore, the decrease of B-50/GAP43 immunoreactivity and the patchy distribution may indicate that synaptogenesis occurred in the glomeruli of the transplant, in spite of the altered topography. Together with previous findings on reinnervation of the piriform cortex from projection neurons situated in olfactory bulb transplants, our immunohistochemical data support the notion that olfactory bulb transplantation may reestablish to some degree the neuronal circuits affected by the experimental surgery.

Increase in glial fibrillary acidic protein following neural trauma

Immunohistochemical staining and quantitative evaluation of glial fibrillary acidic protein (GFAP) were carried out in a stab wound model of neural trauma in the rat. Increased GFAP staining was detected in reactive cortical astrocytes in the vicinity of the wound at 3, 7, and 30 d following injury. Western blots immunostained for GFAP also demonstrated an increase in GFAP in homogenates from the lesioned cortex, compared to the contralateral control side, on days 3, 7, and 30. Specific activity of GFAP expressed as a ratio of lesion/control values showed a fivefold increase from day 0 to day 7, with no further change on day 30. We conclude that neural trauma elicits a quantitative increase in GFAP in the rat cortex during the first week following injury. This increase correlates with both astrocyte hyperthrophy and proliferation. Thus, specific activity of GFAP is a reliable indicator of the onset and progression of astrogliosis in neural trauma.

Turnover of glial filaments in mouse spinal cord.

Twenty-day-old mice received a single tail vein injection of [guanido-14C]arginine. The cytoskeleton was extracted from the spinal cords at varying lengths of time thereafter. Glial fibrillary acidic protein (GFAP) formed a distinct, broad band that was widely separated from other protein bands in one-dimensional polyacrylamide gels. The purity of the GFAP band was verified by Western blot analysis of one- and two-dimensional electrophoretic patterns. In addition, enzyme-linked immunosorbent assay and quantitative Western blot analysis indicated that 95% of the total spinal cord GFAP was extracted in the cytoskeletal preparation. The specific activity of GFAP was obtained by eluting the protein from the cytoskeletal GFAP band in preparative one-dimensional gels. Specific activity reached a peak 2 h after injection with [14C]arginine. Forty percent of the incorporated radioactivity was still present in cytoskeletal GFAP at 9 weeks, indicating that a significant proportion of glial filaments turns over relatively slowly in vivo.