Coexpression of glial fibrillary acidic protein and vimentin in the central and peripheral nervous systems of the twitcher mutant

Chiu, Fung‐Chow; Sacchi, R.; Cláudio, Luz; Kobayashi, S.; Suzuki, Kinuko

doi:10.1002/glia.440010202

Cited by 23 publications

(7 citation statements)

References 36 publications

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“…As previously reported (Chiu et al, 1988), each cytoskel-eta1 pellet, containing 2-3.5 mg protein, was solubilized in 1 ml of 30 mM sodium phosphate buffer, pH 7.5, containing 6 M urea, 2 mM EGTA, 0.5 mM PMSF, 1% (vol/vol) 6-mercaptoethanol, and 1 .O pg/ml leupeptin, and passed through a 0.5-ml column of CM-52 (Whatman Ltd., Kent, England) to remove histones and other low-molecular weight basic proteins. The eluent (approximately 2.5 ml) was injected into a FPLC-Mono Q column (anionic exchanger), 5 X 50 mm (Pharmacia, Piscataway, NJ, U.S.A.), and developed with a 25-ml gradient of 0 to 0.6 MNaCl in the above buffer at a flow rate of 0.5 ml/min.…”

Section: Purification Of Gfap By Fplcsupporting

confidence: 72%

“…Assuming similar findings throughout the length of the spinal cord, this suggests that the decline of vimentin with diminishing inflammation may be balanced by an increase of vimentin in a subpopulation of reactive astrocytes. Others have noted that only a minority of reactive astrocytes in a number of gliotic conditions contain vimentin (Dahl et al, 198 1 ;Pixley and devellis, 1984;Schiffer et al, 1986;Chiu et al, 1988).…”

Section: Discussionmentioning

confidence: 99%

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Glial Fibrillary Acidic Protein Increases in the Spinal Cord of Lewis Rats with Acute Experimental Autoimmune Encephalomyelitis

Aquino

Chiu

Brosnan

et al. 1988

Journal of Neurochemistry

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Glial fibrillary acidic protein (GFAP) in the spinal cords of Lewis rats with acute experimental autoimmune encephalomyelitis (EAE) was quantitated by densitometry of both stained gels and immunoblots of electrophoretically separated cytoskeletal proteins. The experimental period ranged from 7 to 65 days postinoculation (dpi). Greater than 92% of the total spinal cord GFAP was recovered in the Triton-insoluble cytoskeletal pellet; less than 2% was truly soluble. GFAP increased gradually and significantly with time, reaching a level one-and-a-half to two times greater than that of controls by 35 dpi and remaining elevated at 65 dpi. In EAE animals, GFAP was 33% of the total Triton-insoluble protein (excluding histones and other small basic proteins) at 7 dpi, rising to 48% at 65 dpi. Increases in vimentin were also noted, following a time course similar to that of GFAP. An increase in immunocytochemical staining of GFAP was noticeable at 10 dpi and became marked at 14 dpi, a time before GFAP levels had increased significantly. Thus, enhanced staining at the peak of the disease cannot be explained simply by an increase in antigen protein. Other possible explanations, such as an increase in soluble GFAP content, proteolytic degradation, or modifications in the immunochemical properties of GFAP in EAE animals, were ruled out. Both the biochemical and immunocytochemical increases in GFAP persisted through 65 dpi, even though the animals recovered from clinical signs at approximately 18 dpi.

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Section: Purification Of Gfap By Fplcsupporting

confidence: 72%

Section: Discussionmentioning

confidence: 99%

Glial Fibrillary Acidic Protein Increases in the Spinal Cord of Lewis Rats with Acute Experimental Autoimmune Encephalomyelitis

Aquino

Chiu

Brosnan

et al. 1988

Journal of Neurochemistry

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“…We noted previously that vimentin was elevated in chronic relapsing S J W mice (Smith and Eng, 1987), and Aquino et al (1988) found these levels to increase, following a time course similar to that of GFAP. It is likely that some vimentin was contributed by the inflammatory cells (Eng et al, 19861, but co-expression of GFAP and vimentin has been demonstrated in some, but not all, astrocytes from brain of various pathologic conditions (Calvo et al, 1991;Chiu et al, 1988;Dahl et al, 1981;Schiffer et al, 1986). Although vimentin mRNA was not measured here, it is likely to be upregulated on a schedule similar to that of GFAP.…”

Section: Discussionmentioning

confidence: 99%

GFAP mRNA fluctuates in synchrony with chronic relapsing EAE symptoms in SJL/J mice

Kothavale¹,

Gregorio²,

Somera³

et al. 1995

Glia

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Activation of astrocytes and hypertrophy of their processes is a result of a number of pathological conditions in the central nervous system. Astrocytic gliosis is especially prominent in multiple sclerosis (MS), where astrocytic fibers form a dense matrix around demyelinated axons. Experimental allergic encephalomyelitis (EAE), a laboratory model for MS, is also accompanied by astrocytic hyperactivity. We have previously shown the formation of plaque-like structures which stain heavily for glial fibrillary acidic protein (GFAP) in the brains and spinal cords of SJL/J mice after several episodes of chronic relapsing EAE (Smith and Eng: J Neurosci Res 18:203, 1987). To further investigate the mechanisms of this phenomenon, we have measured the levels of mRNA for GFAP throughout the course of three episodes and recoveries of EAE in the SJL/J mouse. Mice were immunized with spinal cord homogenate and subsequently developed EAE. After recovery they were again immunized at appropriate intervals, resulting in successive episodes of EAE, with partial or complete recovery between the paralytic stages. At appropriate times in the course of the different stages of EAE, spinal cords were dissected and RNA was prepared from each spinal cord. RNA was analyzed by Northern blots to determine the levels of mRNA for GFAP and, as a control for the 70 kDa neurofilament (NF-L). With the onset of the first EAE episode GFAP mRNA in spinal cords from animals with mild symptoms increased to sixfold the control level (P < 0.02) and to 20-fold in those with paralysis (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

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“…As the cells of the astrocyte family mature, vimentin is gradually replaced by intermediate filaments composed of the more specific GFAP (Dahl, 1981;Pixley and De Vellis, 1984). In glial cultures (Chiu et al, 1981;Fedoroff, 1986) or under pathological conditions (Chiu et al, 1988;Paulus and Roggendorf, 1988;Pixley and De Vellis, 1984;Sharp et al, 1982), vimentin synthesis may persist or may be resumed, resulting in astrocytes that express both filament proteins simultaneously. It is therefore not surprising that our cultures contained both vimentin single-labeled and vimentidGFAP double-labeled cells and relatively large amounts of both proteins.…”

Section: Discussionmentioning

confidence: 99%

Region‐ and sex‐related differences in maturation of astrocytes in dissociated cell cultures of embryonic rat brain

Beyer

Epp

Fassberg

et al. 1990

Glia

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Previous studies using dissociated cell cultures of fetal rat brain have revealed considerable regional diversity as well as sex steroid-independent sex differences in developmental schedules of dopaminergic neurons. Because these phenomena might be related to glial heterogeneity, cultures of dissociated male and female diencephalon, mesencephalon, and rhombencephalon of gestational day 14 rats were investigated with respect to the development of astrocytic markers. Cultures were incubated for 3-8 days in vitro (DIV) in serum-supplemented or serum-free medium. Vimentin and glial fibrillary acidic protein (GFAP) were quantified by counting of immunolabeled cells and immunoblotting. Vimentin and GFAP content rose from DIV 3 to 6 in all cultures. Regional variation of vimentin content was low, but large differences occurred in amounts of GFAP. GFAP reached high levels in rhombencephalon, especially when supplemented with serum, but remained very low or not detectable in mesencephalon. Simultaneous immunostaining for both cytoskeletal proteins revealed the presence of large numbers of vimentin single-labeled and small numbers of vimentin/GFAP double-labeled cells. Numbers of cells expressing GFAP showed similar regional variations as GFAP contents in both serum-free and serum-supplemented medium. They rose steeply from DIV 3 to 8 in rhomb- and diencephalon but not in mesencephalon. Transiently, female diencephalic cultures contained slightly more GFAP-immunoreactive cells than male cultures. The results thus demonstrate considerable regional heterogeneity of astrocytic maturation. However, neither the regional nor the sex differences show a consistent correlation with previous data on development of dopaminergic and other monoaminergic neurons in vitro. It seems likely that the dependence of neurons on glial environment for realization of an inherent developmental program varies among neuronal phenotypes.

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Coexpression of glial fibrillary acidic protein and vimentin in the central and peripheral nervous systems of the twitcher mutant

Cited by 23 publications

References 36 publications

Glial Fibrillary Acidic Protein Increases in the Spinal Cord of Lewis Rats with Acute Experimental Autoimmune Encephalomyelitis

Glial Fibrillary Acidic Protein Increases in the Spinal Cord of Lewis Rats with Acute Experimental Autoimmune Encephalomyelitis

GFAP mRNA fluctuates in synchrony with chronic relapsing EAE symptoms in SJL/J mice

Region‐ and sex‐related differences in maturation of astrocytes in dissociated cell cultures of embryonic rat brain

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