OBJECTIVETo study the potential interaction between PrP protein and glial fibrillary acidic protein (GFAP) and identify the binding region within PrP with GFAP.

METHODSThe supernatant of healthy and scrapie-infected hamsters' brain homogenate was prepared, while various recombinant PrP or GFAP proteins were expressed using prokaryotic-expressing or in-vitro translation system. The possible molecular interaction between PrP proteins and GFAP was tested by Pull-down and immunoprecipitation assays.

RESULTSBoth native PrP(C) and its protease-resistant isoform (PrP(Sc)) formed complexes with the native GFAP. The full-length recombinant PrP proteins interacted with GFAP. The domain responsible for interacting GFAP was located at C-terminal of PrP (residues 91 to 231).

CONCLUSIONThe studies of the association of PrP with GFAP may further provide insight into a potential role of GFAP in the biological function of PrP and the pathogenesis of prion disease.

Animals ; Brain ; metabolism ; Cricetinae ; Gene Deletion ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Immunoprecipitation ; Mice ; Prions ; genetics ; metabolism ; Protein Binding ; Recombinant Proteins ; metabolism

OBJECTIVETo investigate whether gliosis in the brain tissues of the hamsters infected with various amounts of scrapie strain 263K is correlated with the inoculation doses or the incubation times.

METHODSThe total values of glial fibrillary acidic protein (GFAP) in brains were evaluated by Western Blots and the GFAP-stained cells were detected by immunohistochemistry (IHC). The characteristics of GFAP distributions among various groups were defined by quantitive and statistic analyses.

RESULTSCompared with the brain tissues of normal hamsters, remarkably higher total GFAP levels and more GFAP-stained cells were observed in the brain tissues of infected ones, howbeit, no significant difference was addressed among the infected groups.

CONCLUSIONInoculations of various amounts of scrapie strain 263K into experimental hamsters intracerebrally induced the similar patterns of gliosis in the brains at the clinically terminal stage, regardless of infectious doses and incubation times.

Animals ; Brain ; metabolism ; pathology ; Cricetinae ; Gene Expression ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Gliosis ; metabolism ; pathology ; Humans ; PrPSc Proteins ; metabolism ; Prion Diseases ; metabolism ; pathology

Animals ; Epilepsy ; genetics ; Gene Expression Regulation ; drug effects ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Hippocampus ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Scorpion Venoms ; pharmacology

Alzheimer Disease ; pathology ; China ; Genes, p53 ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Multiple Sclerosis ; pathology ; Nervous System Neoplasms ; classification ; genetics ; metabolism ; pathology

OBJECTIVETo study the effect of melatonin on the expressions of glial fibrillary acidic protein (GFAP), nuclear factor-κB (NF-κB p65) and synaptophysin in mice of different ages.

METHODSTwenty young male B6C3F1 mice (5.5 months) and 20 aged mice (26 months) were both divided into control and melatonin treatment (daily dose of 0.04 mg/kg) groups. After 2.5 months of treatment, the brain tissues of the mice were collected to examine the expressions of GFAP, NF-κB and SYN by immunohistochemistry.

RESULTSIn the control groups, the expression of NF-κB p65 in the brain tissue increased with age, whereas a reverse change was found in melatonin-treated aged rats (P<0.05). Synaptophysin expression also decreased with age, but melatonin treatment significantly enhanced its expression in aged mice (P<0.05). GFAP expression in the brain tissue increased with age regardless of melatonin treatment (P>0.05).

CONCLUSIONGFAP expression is almost not affected by melatonin treatment in aged mice. Melatonin can reduce the expression levels of NF-κB p65 and synaptophysin in the brain tissue to protect the brain and slow down the aging process.

Aging ; metabolism ; Animals ; Brain ; metabolism ; Chimera ; Glial Fibrillary Acidic Protein ; Male ; Melatonin ; pharmacology ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; NF-kappa B ; genetics ; metabolism ; Nerve Tissue Proteins ; genetics ; metabolism ; Synaptophysin ; genetics ; metabolism ; Transcription Factor RelA ; genetics ; metabolism

OBJECTIVETo explore the changes in the expressions of glial fibrillary acidic protein (GFAP) and growth- associated protein-43 (GAP-43) in retinal ganglial cells after neural transplantation.

METHODSThirty-nine rats were randomized into normal control group, nerve amputation group and nerve amputation with peripheral nerve transplantation group. Immunohistochemistry was used to detect the changes in the expressions of GFAP and GAP-43 at different time points after the operations, and real-time PCR was employed to detect the mRNA expressions of 13 genes in the retinal ganglial cells of the rats.

RESULTSImmunohistochemistry showed obviously increased GFAP expressions in the retina following the nerve amputation. GFAP expression was down-regulated while GAP-43 expression upregulated in the retinal ganglial cells after peripheral nerve transplantation. Real-time PCR results showed that 5 days after the operations, retinal GFAP and GAP-43 expressions increased significantly in the nerve amputation group and peripheral nerve transplantation groups as compared with those in the control group, but GAP-43 expression decreased significantly in the former two groups afterwards.

CONCLUSIONThe regenerated retina may adjust the production of GFAP. The retinal ganglial cells express GAP-43 during retinal regeneration. Up-regulation of the expression of GAP-43 provides the evidence for nerve regeneration following the nerve transplantation.

Animals ; Axons ; Female ; GAP-43 Protein ; genetics ; metabolism ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Nerve Regeneration ; genetics ; Optic Nerve ; transplantation ; Optic Nerve Injuries ; metabolism ; Random Allocation ; Rats ; Retinal Ganglion Cells ; metabolism

BACKGROUNDMüller cells in the mammalian retina normally express low levels of glial fibrillary acidic protein (GFAP); however, its expression is upregulated in response to the loss of retinal neurons. The change in expression of GFAP is one of the earliest indicators of retinal damage and is correlated with the time course of disease. The aim of this study was to investigate the time course of degeneration and the expression of GFAP in the retina of mer knockout mice.

METHODSA total of 30 mer knockout mice, aged from 15 - 20 days to 1 year and 32 age-matched wild type mice as controls were tested. Immunohistochemistry was used to show the expression of GFAP in the central and peripheral retina of mer knockout and control mice at postnatal age of 15 days (P15d), 20 days (P20d), 4 weeks (P4w), 6 weeks (P6w), 8 weeks (P8w), 3 months (P3m), 6 months (P6m) and 1 years (P1y).

RESULTSThe expression of GFAP in the central and peripheral retina of wild type mice was limited to the retinal ganglion cell and nerve fiber layers. In the central retina of mer knockout mice, GFAP expression was upregulated at P4w and GFAP immunolabelling penetrates across the entire thickness of the retina at P8w; whereas in the peripheral retina, the GFAP expression was upregulated at P20d and GFAP immunolabelling penetrates the entire retina after P4w.

CONCLUSIONSIncreased expression of GFAP in Müller cells of mer knockout mice occur at P20d in the peripheral retina and P4w in the central retina. GFAP expression in Müller cells appears to be a secondary response to the loss of retinal neurons. Increased expression of GFAP may occur prior to any detectable morphological changes in the retina. This study suggests that the loss of retinal neurons may begin in the early stages of retinitis pigmentosa, prior to the discovery of any morphological changes in the retina.

Animals ; Glial Fibrillary Acidic Protein ; metabolism ; Immunohistochemistry ; Mice ; Mice, Knockout ; Proto-Oncogene Proteins ; genetics ; Receptor Protein-Tyrosine Kinases ; genetics ; Retina ; metabolism ; pathology ; Retinitis Pigmentosa ; genetics ; metabolism ; c-Mer Tyrosine Kinase

OBJECTIVETo observe the dose-effect relationship of electroacupuncture of different current intensities combined with Morphine of different dosage on alleviating the rats' tibial cancer pain, and explore the possible mechanism, which could provide the experiment basis for alleviating the tibial cancer pain by electroacupuncture combined with Morphine.

METHODSOne hundred female Wistar rats were randomly divided into a normal group, a model group and eight treatment groups, 10 cases in each group. The rats in the treatment groups were treated by combined therapies of electroacupuncture of different intensities with 2 Hz /100 Hz dense-disperse wave on "Jiaji"(EX-B 2)and different dosage Morphine in 2 factor 3 level conditions, once a day for 6 days. The pain thresholds were observed before the treatment and 0 min, 1 h, 2 h and 5 h after the first treatment as well as after 3 and 6 times of treatments. The glial fibrillary acidic protein (GFAP) expression was determined by immunohistochemical method.

RESULTSThe rats' pain thresholds were significantly increased with electroacupuncture of 2 mA and 1 mA (all P < 0.01) on the 0 min, 1 h and 2 h of the first treatment, between which there were no significant differences (all P > 0.05). The pain threshold was still increased by electroacupuncture of 2 mA on the 5 h of the treatment (P < 0.01), while that of 1 mA failed to take effect (P > 0.05). After 3 and 6 times of treatments, both electroacupuncture of 2 mA and 1 mA had the effect of increasing the pain threshold (all P < 0.01), and the effect of 2 mA was superior to that of 1 mA (P < 0.05), had the synergistic effect with 5 mg/(kg x d) Morphine (P < 0.05). After 6 times of treatments, both electroacupuncture of 2 mA and 1 mA could inhibit the expression of GFAP (both P < 0.01), and there was no significant difference between them (P > 0.05). Both of 5 mg/(kg x d) and 2.5 mg/(kg x d) of Morphine, however, didn't bring about inhibition effect (P > 0.05).

CONCLUSIONThere is a does-effect relationship on electroacupuncture of different current intensity for alleviating the tibial cancer pain in rats. The electroacupuncture with 2 mA, which is better than that with 1 mA, has the synergistic effect with 5 mg/(kg x d) of Morphine. The electroacupuncture can inhibit the expression of GFAP to cooperate with Morphine for the purpose of alleviating the rats' tibial cancer pain.

Animals ; Bone Neoplasms ; complications ; genetics ; metabolism ; Electroacupuncture ; instrumentation ; methods ; Female ; Gene Expression Regulation, Neoplastic ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Humans ; Pain ; etiology ; genetics ; metabolism ; Pain Management ; instrumentation ; methods ; Rats ; Rats, Wistar ; Spine ; metabolism ; Tibia ; metabolism

OBJECTIVETo establish differential proteomics profiles of glioblastoma cell lines from Chinese, and to provide reference for future basic studies.

METHODSTotal protein was extracted from 3 glioblastoma cell lines, CHG-5, TJ899 and TJ905. After normalization, the total protein was presented by two-dimensional (2D) electrophoresis, scanned and analyzed. Some of the identified protein spots were verified by immunocytochemistry of cell lines and immunohistochemistry of solid tumors. The glia cells were used as the control throughout the study.

RESULTSA total of 13 differential protein spots were selected, and eventually 10 were identified as unique proteins. These 10 proteins were involved in cytoskeleton forming, cellular metabolism, tumor migration, stress and inflammatory reaction. Immunocytochemistry and immunohistochemistry further confirmed these proteins present in the solid tumors.

CONCLUSIONDistinct differential proteomics profiles exist in glioblastoma cell lines and normal glia cells, likely related to the transformation of normal glia to glioma.

Brain Neoplasms ; genetics ; metabolism ; Cathepsin D ; metabolism ; Cell Line, Tumor ; Gene Expression Profiling ; Glial Fibrillary Acidic Protein ; metabolism ; Glioblastoma ; genetics ; metabolism ; Humans ; Microfilament Proteins ; metabolism ; Neuroglia ; metabolism ; Proteomics ; methods

OBJECTIVETo investigate the expression and significance of peroxisome proliferators-activated receptor gamma (PPAR gamma) in human glioma.

METHODSImmunohistochemical staining for PPAR gamma was performed using biopsy specimens of human glioma of various histological types. Expression of PPAR gamma and GFAP in glioma cell lines SWO-38, U251 and SHG-44 were analyzed using Western blotting and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSImmunohistochemical study showed that PPAR gamma was expressed in glioma tissues with positive rate of 37.5%. Western blotting and RT-PCR showed that PPAR gamma was expressed in both glioma cell lines SWO-38 and U251, but not in SHG-44 cells. However, high expression of GFAP was detected in SHG-44 cells.

CONCLUSIONPPAR gamma is associated with carcinogens of glioma. Actived PPAR gamma by agonist may be a novel approach to the treatment of glioma.

Blotting, Western ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Glial Fibrillary Acidic Protein ; biosynthesis ; genetics ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; PPAR gamma ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction