Glial-like cells of the rat pituitary intermediate lobe change morphology and shift from vimentin to GFAP expression during development

This study demonstrated morphological changes in glial-like cells of the rat pituitary intermediate lobe during early postnatal development, and a subsequent shift in protein expression from vimentin to GFAP. Vimentin immunoreactivity was detected in the lobe at embryo day 14 and was localized in radially-oriented, bipolar cells whose processes spanned the thickness of the intermediate lobe. At electron microscopical resolution, processes contained intermediate filaments, cell nuclei were indented while secretory vesicles characteristic of the endocrine cells were not found. Vimentin immunoreactive intensity began to decrease at postnatal day 5. By postnatal day 7, vimentin-positive, setellate cells were observed, with few radial processes found by day 10. The intensity of vimentin immunoreactivity decrease through day 25. Within the lobe parenchyma, vimentin was localized in glial-like cells since double-label immunohistochemistry revealed no colocalization of β-endorphin and vimentin, or fibronectin and vimentin. Dopamine-containing axons were in close apposition to vimentin-positive processes. GFAP immunoreactivity first appeared on postnatal day 20 and, by day 25, stellate cell bodies with three to six extended processes were evident. Cells were primarily distributed in the caudal third of the lobe. The characteristic adult pattern of cell clusters in latero-dorsal and ventral portions of the lobe was fully established by postnatal day 55. The transition from vimentin to GFAP expression and concurrent morphological changes resemble those described for radial glia during cerebral cortical development.     

Immunohistochemical observations on rat radial glia: relationship with the origin of ethylnitrosourea-induced tumors

Summary Gliomas induced in the rat by transplacental administration of ethylnitrosourea (ENU) are intensely immunoreactive for vimentin and scarcely for glial fibrillary acidic protein (GFAP). Since tumoral transformation takes place during the late fetal and early postnatal period, the sequential expression of the two glial antigens has been investigated in this age period in ENU-treated and control rats. Immunohistochemical and immunoelectron microscopical methods have been employed. Vimentin was widely expressed starting from embryonal day 14 (E 14) in the processes of radial glia; as long as radial glia was present, vimentin decorated it. GFAP was, at earliest, observed at E 20 and expressed by glial cells with a stellate, i.e., mature shape. No GFAP-positive radial process was observed. No difference was found between ENU-treated and control rats. Since ENU is most effective in producing tumors when administered at the 16–17th day of fetal life, vimentin-positive radial glia is a candidate target of ENU. The similarity of intermediate filament pattern between radial glia in the late fetal life and tumors induced by transplacental ENU suggests that radial glia might be the cell of origin.Supported in part by M. P. I. 60% grant, Rome     

Development of glial cells in the cerebral wall of ferrets: direct tracing of their transformation from radial glia into astrocytes

Coronal sections of the cerebral wall from developing ferrets (newborn to adult) were double-stained with antibodies to vimentin and glial fibrillary acidic protein (GFAP). At birth, the dominant glial population was radial glia and these cells labeled only for vimentin. A small population of immature astrocytes in the cortical plate was double labeled for GFAP and vimentin. In successive days, the number of vimentin-positive radial glia gradually decreased and they disappeared entirely at about 21 days. During this same period, the double-stained astrocytes increased in number and were distributed throughout the cortical plate and intermediate zone. After 6 weeks of age the astrocytes were mostly confined to the developing white matter. Around this time they gradually lost their vimentin staining, and in the adult no vimentin-positive elements were seen except at the ependymal surface. In newborn ferrets single radial glial cells were also visualized by applying the carbocyanine dye DiI onto the pial surface of fixed brains. While most radial glia extended from the ventricular zone to the pial surface, a substantial fraction of them had lost their contact to the ventricular zone. Their somata were displaced into the subventricular zone and lower portion of the intermediate zone. The possibility that radial glia transform into astrocytes was directly tested by injecting fluorescent dyes under the pial surface of newborn ferrets at a time when virtually no GFAP-positive astrocytes are present. The tracer, which was taken up in the upper portion of the cortical plate, stained the radial glial cell somata in the ventricular zone in a similar way as the dye DiI did in the fixed brains. As the radial glial cells disappeared at successively longer survival times, the tracer was ultimately found within newly formed GFAP-positive astrocytes. These results provide strong support for the hypothesis that radial glia cells are the immature form of astrocytes (Choi and Lapham: Brain Res. 148:295-311, '78; Schmechel and Rakic: Anat. Embryol. (Berl.) 156:115-152, '79), and also show that, at least in the ferret cortex, the transformation is accompanied by a change in the expression of intermediate filament protein.     

Localization of mu class glutathione-S-transferase in the forebrains of neonatal and young rats: implications for astrocyte development.

The Yb (Mu class) isoform of glutathione-S-transferase has recently been localized in ependymal cells, subependymal cells, and astrocytes in the forebrains of rats 3 weeks to adult in age. It was not known, however, at what age Mu might first be observed during postnatal development and whether the first cells in which it was found would be immature astrocytes or some less differentiated glial precursor cell, if the latter were present in vivo. Tissue sections from the forebrains of neonatal to 16 day old rats were immunostained with antibodies against Mu. In neonates Mu was observed in vimentin-positive cells and their processes adjacent to the lateral ventricles, and in the corpus striatum. The colocalization with vimentin suggested that these were subependymal cells and radial glia. In the corpus striatum the radial glia, while still vimentin-positive, rapidly lost Mu from their radial cell processes, whereas the cell-bodies remained Mu-positive. During the first postnatal week the Mu-positive, glial-fibrillary-acidic-protein (GFAP)-positive cell bodies of immature astrocytes appeared in the corpus striatum. The earliest Mu-positive cells in the immature white matter of the corpus callosum were vimentin-positive and had striking longitudinal processes that also were vimentin- and Mu-positive. Like the processes of radial glia, the longitudinal processes lost their Mu-immunoreactivity, only later and more gradually. Mu-positive, GFAP-positive cells appeared later in the corpus callosum than in the corpus striatum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Radial Glia-Like Cells in the Supraoptic Nucleus of the Adult Rat

Conventional light and confocal microscopy of thick vibratome sections of the hypothalamus of adult male and female rats immunostained for the astrocytic marker glial fibrillary acidic protein (GFAP) revealed that the supraoptic nucleus (SON) contains two morphologically distinct types of astrocytes. One has a stellate form, similar to that of most astrocytes in the adult CMS. The other has a morphology reminiscent of radial glia in the developing CNS: from their cell bodies, located along the ventral glia lamina (VGL), arise one long thick process that spans the SON in the coronal plane, several horizontally-oriented processes that form a dense network in the VGL, and a short process oriented towards the pia. The latter astrocytes are immunoreactive for vimentin, an intermediate filament protein of immature glial cells and a marker for radial glia. The stellate astrocytes showed no vimentin immunoreactivity. The functional significance of each type of supraoptic astrocyte is at present unknown but the presence of radial glia-like cells in this hypothalamic region suggests that the SON retains a certain degree of immaturity during adulthood, that may be linked to its well known capacity to undergo neuronal-glial plasticity under physiological and experimental stimulation.     

Glial environment in the developing superior colliculus of hamsters in relation to the timing of retinal axon ingrowth

We have examined the developmental changes of glial cell organization in the superior colliculus of embryonic and neonatal hamsters in reference to the known sequence of retinal axon ingrowth and arborization in the midbrain. Immunolocalization of vimentin, a marker for neuronal and glial cell precursors, reveals a uniform distribution of radially oriented cells, with perikarya located at the ventricular surface and thin, elongated processes fanning out toward the pia. These vimentin-positive cells, referred to as the lateral radial cells, are present in the tectum from embryonic day (E) 10 (earliest day examined) until approximately postnatal day (P) 5. Vimentin expression in the lateral radial cells decreases markedly during the second week of postnatal life: application of DiI to the ventricular surface reveals that the pial attachment of the lateral radial cells is withdrawn and that the radial processes are gradually pulled back toward the ventricular zone. By P14, virtually no vimentin-positive radial cells are detectable in the superior colliculus. At no time during development are the lateral radial cells immunopositive for the glial fibrillary acidic protein (GFAP); however, shorter, vimentin-positive astrocytic profiles can be seen in the tectum, around the time the radial fibers have been withdrawn, suggesting that at least some radial cells are transformed into astrocytes that will colonize the mature colliculus. At approximately E12, a second group of cells, referred to as the midline radial glia, is detected at the tectal midline. These cells are tightly bundled, forming a raphe in the tectum. They are intensely vimentin positive from E13 until at least P14. From the time of birth, the midline radial cells also exhibit intense immunoreactivity for GFAP. The lateral radial cells are present in the superior colliculus prior to and during the period of neurogenesis but remain well past the time when collicular neuronal migration is completed. Pial processes of the lateral radial cells are present within the superficial tectal layers during the time retinal axons are entering this target; they may be involved in directing the growth and initial collateralization of retinotectal axons. Their withdrawal from retinorecipient collicular zones begins at about the time arbors are being elaborated on retinal axons. In constrast, the midline glia become distinct just prior to the time retinal axons enter the superior colliculus and persist during the time retinotectal projections are being fully established. These raphe glia may be involved in maintaining the laterality of the retinotectal projection. © 1995 Wiley-Liss, Inc.     

Sexual dimorphism in the hamster cerebellum demonstrated by glial fibrillary acidic protein (GFAP) and vimentin immunoreactivity.

Male and female hamsters aged 1, 4, and 10 postnatal weeks were used to study the distribution of vimentin and glial fibrillary acidic protein (GFAP) in the cerebellum. Vimentin immunoreactivity exceeded that of GFAP during the first postnatal week, although GFAP was also observed in all cerebellar layers. Immunoperoxidase analysis revealed that by the fourth postnatal week vimentin was only detected in Bergmann fibers and the very scarce fibrous astrocytes located in the inner white matter. The Purkinje cell bodies were only coated with GFAP-immunopositive processes. At 10 weeks, vimentin immunoreactivity was reduced to thin Bergmann glial processes, whereas GFAP immunoreactivity had greatly increased in the whole cerebellum. The GFAP immunostaining was denser in males than in females; however, in females, the Bergmann fibers were heavily immunostained with anti-vimentin in contrast to the males. The results described in the present paper indicate a sex difference in vimentin and GFAP immunoreactivities in the cerebellar astrocytes at 4 weeks of age, which persisted in the oldest hamsters in this study. The existence of sexual dimorphism might suggest that the expression of both gliofilament proteins could be influenced by circulating sex steroids.     

Effect of thyroid deficiency on the development of glia in the hippocampal formation of the rat: an immunocytochemical study

The development of glia in the hippocampal formation of normal and hypothyroid rats was studied using immunocytochemical staining for either glial fibrillary acidic protein (GFAP) or vimentin. Light microscopy showed lower GFAP immunoreactivity in the radial glial processes of young hypothyroid rats compared to normal animals. These processes followed the known path of neuroblast migration toward the proliferative zone of the dentate gyrus until the end of the 1st postnatal week. Vimentin immunoreactivity showed that the glial processes were present and therefore immature at least with respect to their cytoskeletal composition. We propose that this early defect in the maturation of the radial glial fibers accounts for the final deficit in the granule cells of the dentate gyrus. Later in development, thyroid deficiency also reduced the density and number of GFAP-labeled astrocytes and the growth of their processes. This observation is in complete disagreement with the glial hypertrophy induced by thyroid deficiency in the cerebellum. The considerably increased histogenetic cell death observed in the cerebellum of young hypothyroid rats could in turn induce glial hypertrophy, whereas the hippocampal formation, where a normal low number of cell deaths is observed, is only subjected to the general depressive effect of thyroid deficiency on cell maturation.     

Development of astroglial elements in the suprachiasmatic nucleus of the rat: with special reference to the involvement of the optic nerve

The development of astroglial cells and the effect of the retinohypothalamic tract on it were studied by vimentin and glial fibrillary acidic protein (GFAP) immunocytochemistry in the suprachiasmatic nucleus (SCN) of the rat. At the embryonic stage, vimentin-immunoreactive (VIM-IR) radial glia, precursors of astrocytes, were dominant. However, their filaments vanished in the first few postnatal days. Instead of VIM-IR glial filaments, GFAP-immunoreactive (GFAP-IR) astrocytes appeared at E20 and grew rapidly from the P3 stage. GFAP immunoreactivity in the ventrolateral portion of the SCN (VLSCN) was measured using a computer-assisted image analyzing system. In normal rats, GFAP immunoreactivity showed a stepwise pattern with two slopes at P3-P4 and P20-P25. Bilaterally eye-enucleated rats operated on the day of birth showed lower GFAP immunoreactivity than normal rats and the GFAP immunoreactivity did not increase between P20 and P25 when GFAP-IR glial processes rapidly expand. Electron microscopic investigation at P50 (adult stage) revealed that neurons in the VLSCN had often direct apposition without astroglial processes and the frequency of this finding was significantly higher in eye-enucleated rats than in the control rats. These findings strongly suggest that the postnatal development of astroglial elements, particularly the expansion of GFAP-IR processes in the SCN, is regulated by retinohypothalamic projection.     

Co-expression of glial fibrillary acidic protein- and vimentin-type intermediate filaments in human astrocytomas

Summary The expression of intermediate filament (IF) proteins was studied in 71 cases of malignant human astrocytoma and in 17 cases of reactive gliosis, using immunocytochemical techniques with polyclonal and monoclonal antibodies to glial fibrillary acidic protein (GFAP) and vimentin. In all cases of astrocytoma, varying in degree of malignancy from grade I to grade IV, co-expression of GFAP and vimentin was found. No change in vimentin- or GFAP-IF expression with increasing anaplasia was seen. In addition astrocytic cells in reactive gliosis showed simultaneous expression of GFAP and vimentin. The intracellular distribution of these IF proteins differed. Vimentin was found to be located in a more juxta-nuclear position, whereas GFAP immunoreactivity showed a more intense staining of the cellular processes. Astrocytes in reactive gliosis behaved more or less like neoplastic cells. However, thin cell processes of reactive astrocytes in the cortex and superficial white matter only contained GFAP immunoreactivity. Simultaneous expression of GFAP and vimentin and their proportion in malignant and reactive glial cells are discussed in the light of earlier reports on the IF content of glial cells during development and maturation, in which vimentin precedes GFAP-expression. The existence of two separate (functional) IF systems in astroglia is suggested.     

Vimentin: changes in distribution during brain development

This paper examines both the anatomical changes in the distribution of vimentin intermediate filament protein and the biochemical changes in vimentin and its degradative enzyme during postnatal brain development in the tree shrew (Tupaia belangeri). A pattern of multiple immunoreactive bands at birth (postnatal day 0, or P0) was revealed in nitrocellulose blots of polyacrylamide gels ("Western blots"). These multiple bands gradually disappear during development, and in the adult a single band at the published molecular weight for vimentin (57 kD) is seen. This pattern of bands probably reflects shifts in the activity of a calcium-activated vimentin protease. The changes in the anatomical distribution of vimentin-immunoreactive (vimentin+) cells and their fine processes parallel the biochemical shifts seen in immunoblots. We have examined the neocortex, lateral geniculate nucleus (LGN), and hippocampus in detail. During the first postnatal week, vimentin+ glia, especially radial glia, are prominent in both neocortex and hippocampus. In contrast, only a few vimentin+ radial glia remain in the thalamus at this age. Vimentin+ glia appear to coincide with bundles of axons and often seem to outline subdivisions of thalamic nuclei. Additionally, cellular layers of the lateral geniculate nucleus (LGN) appear to stain with antibodies to vimentin several days before the characteristic neuronal cell layers appear in this area. During the second postnatal week, vimentin+ cells appear in "patches" throughout the cortex. Some subdivisions of the thalamus and hippocampus (as defined by cytoarchitectonic differences in the adult) are distinct when the tissue is stained with an antibody to vimentin, even though a conventional Nissl stain at this age shows no apparent delineation in these same regions. Finally, in the adult, only a few vimentin+ cells remain, primarily in the white matter. Taken together, these results suggest that the remodeling of vimentin+ intermediate filaments in immature glial cells (including radial glia) is paralleled by the action of the enzyme which breaks down these filaments. The apparent activity of this enzyme is high early in development as radial and other glia are rapidly dividing and undergoing morphological changes, with a decrease in activity in the juvenile and adult brain, as immature glial cells are supplanted by mature forms.     

Organization of radial and non-radial glia in the developing rat thalamus

The organization of glia and its relationship with migrating neurons were studied in the rat developing thalamus with immunocytochemistry by using light, confocal, and electron microscopy. Carbocyanine labeling in cultured slice of the embryonic diencephalon was also used. At embryonic day (E) 14, vimentin immunoreactivity was observed in radial fascicles spanning the neuroepithelium and extending from the ventricular zone to the lateral surface of the diencephalic vesicle. Vimentin-immunopositive fibers orthogonal to the radial ones were also detected at subsequent developmental stages. At E16, radial and non-radial processes were clearly associated with migrating neurons identified by the neuronal markers calretinin and gamma-aminobutyric acid. Non-radial glial fibers were no longer evident by E19. Radial fibers were gradually replaced by immature astrocytes at the end of embryonic development. In the perinatal period, vimentin immunoreactivity labeled immature astrocytes and then gradually decreased; vimentin-immunopositive cells were only found in the internal capsule by the second postnatal week. Glial fibrillary acidic protein immunoreactivity appeared at birth in astrocytes of the internal capsule, but was not evident in most of the adult thalamic nuclei. Confocal and immunoelectron microscopy allowed direct examination of the relationships between neurons and glial processes in the embryonic thalamus, showing the coupling of neuronal membranes with both radial and non-radial glia during migration. Peculiar ultrastructural features of radial glia processes were observed. The occurrence of non-radial migration was confirmed by carbocyanine-labeled neuroblasts in E15 cultured slices. The data provide evidence that migrating thalamic cells follow both radial and non-radial glial pathways toward their destination.     

Differentiation of radial glia from radial precursor cells and transformation into astrocytes in the developing rat spinal cord

Radial glial cell origins and functions have been studied extensively in the brain; however, questions remain relating to their origin and fate in the spinal cord. In the present study, radial glia are investigated in vivo using the neuroepithelial markers nestin and vimentin and the gliogenic markers GLAST, BLBP, 3CB2, and glial fibrillary acidic protein (GFAP). This has revealed heterogeneity among nestin/vimentin-positive precursor cells and suggests a lineage progression from neuroepithelial cell through to astrocyte in the developing spinal cord. A population of self-renewing radial cells, distinct from an earlier pseudo-stratified neuroepithelium, that resemble radial glial cells in morphology but do not express GLAST, BLBP, or 3CB2, is revealed. These radial cells arise directly from the spinal cord neuroepithelium and are probably the progenitors of neurons and the earliest appearing radial glial cells. GLAST/BLBP-positive radial glia first appear in the ventral cord at E14, and these cells gradually transform through one or more intermediate stages into differentiated astrocytes. Few if any neurons appear to be derived from radial glial cells, which are instead the major sources of astrocytes in the spinal cord. Evidence for the nonradial glial cell origins of some white matter astrocytes is also presented.     

Glial fibrillary acidic protein (GFAP) in oligodendrogliomas: a reflection of transient GFAP expression by immature oligodendroglia

Fourteen pure oligodendrogliomas were studied by light- and electronmicroscopy and immunohistochemistry to examine glial fibrillary acidic protein (GFAP) positivity in the tumors. To compare the immunohistochemical staining patterns of neoplastic oligodendroglia and immature oligodendroglia, myelination glia in the white matter of eight normal brains from children under 6 months of age were studied. The tumors possessed light microscopic and ultrastructural features characteristic of oligodendrogliomas. Microtubules were found in the cytoplasm of nine tumors on electronmicroscopy. In one, intermediate filaments and microtubules were observed in occasional tumor cells with polygonal crystalline structures in the cytoplasm. Using the peroxidase-antiperoxidase technique, all specimens were stained for GFAP, vimentin, S-100 and neuron-specific enolase (NSE). In nine tumors, variable numbers of cells with an oligodendroglial morphology reacted positively for GFAP. All tumors were positive for S-100 and negative for vimentin and NSE. The myelination glia in the eight normal brains stained positively for GFAP but not for vimentin. Vimentin is expressed by developing, reactive and neoplastic astrocytes. Thus, GFAP positivity combined with vimentin negativity in both neoplastic and immature oligodendroglia suggests that GFAP positivity in oligodendrogliomas may reflect the transient expression of this intermediate filament by immature oligodendroglia.     

Gliogenesis in rat spinal cord: evidence for origin of astrocytes and oligodendrocytes from radial precursors

We have examined glial cell lineages during rat spinal cord development by using a variety of antibodies that react with immature and mature glia. Radial glia in embryonic cord bound 1) A2B5, an antibody that reacts with a glial precursor cell population in optic nerve; 2) AbR24, which is directed against GD3 ganglioside and binds to immature neuroectodermal cells and to developing oligodendrocytes in forebrain and cerebellum; and 3) an antibody to the intermediate filament, vimentin. With time, two different populations emerged, both of which seemed to be derivatives of radial cells. One cell type expressed the astrocyte intermediate filament, GFAP, in addition to vimentin. GFAP-containing cells eventually took on the forms of astrocytes in gray and white matter. The other type expressed carbonic anhydrase, an enzyme characteristic of oligodendrocytes and enriched in myelin. Carbonic anhydrase-positive cells eventually developed into small cells with oligodendrocyte morphology. Our observations suggest a common lineage for astrocytes and oligodendrocytes from radial cells during spinal cord gliogenesis.     

Carbonic anhydrase in distinct precursors of astrocytes and oligodendrocytes in the forebrains of neonatal and young rats.

Carbonic anhydrase is present in oligodendrocytes and astrocytes in the mature rat brain. Whereas carbonic anhydrase-positive oligodendrocyte precursors had been identified during the first postnatal week, no information was available about the earliest occurrence of carbonic anhydrase in the astrocytic cell line, nor had carbonic anhydrase been detected in astrocytes in neonatal rat brains. Beginning on the first postnatal day, rat brains were double immunostained with anti-carbonic anhydrase II and respective 'markers' for immature and mature astrocytes and oligodendrocytes. During the first postnatal week there were intensely carbonic anhydrase-positive cells which were ovoid or had broad processes. On the basis of their shapes and antigen contents these were considered to be precursors of oligodendrocytes. Beginning on the first postnatal day carbonic anhydrase II was also observed in some vimentin-positive radial glia and in other vimentin-positive cells that differed in their appearance from the immature oligodendrocytes. The vimentin-positive, carbonic anhydrase-positive cells were less smooth-surfaced, and had much finer processes, than the oligodendrocyte precursors. By the third postnatal day there appeared carbonic anhydrase-positive, glial fibrillary acidic protein (GFAP)-positive cells that resembled the vimentin-positive cells. It is concluded that the latter are immature astrocytes and that carbonic anhydrase is in distinct precursors of oligodendrocytes and astrocytes as early as the first postnatal day.     

Development of vimentin and glial fibrillary acidic protein immunoreactivities in the brain of gray mullet (Chelon labrosus), an advanced teleost

Previous studies in teleosts have revealed the presence of the intermediate filaments vimentin (Vim) and glial fibrillary acidic protein (GFAP) in glial cells of the spinal cord and/or some brain regions, but there is no comprehensive study of their distribution and developmental changes in fishes. Here, the distribution of Vim and GFAP immunoreactivities was studied in the brain of larvae, juveniles, and adults of an advanced teleost, the gray mullet (Chelon labrosus). A different sequence of appearance was observed for expression of these proteins: Vim levels decreased with age, whereas GFAP increased. In general, both immunoreactivities were expressed early in perikarya and endfeet of ependymocytes (tanycytes), whereas expression in radial processes appeared later. In large larvae, the similar expression patterns of Vim and GFAP suggest that some of these glial cells contain both proteins. Subependymal radial glia cells were observed mainly in the optic tectum, exhibiting Vim and GFAP immunoreactivity. The only immunoreactive cells with astrocyte-like morphology were observed in the optic chiasm of the adult, and they were positive for both GFAP and Vim. The perivascular processes of glial cells showed a different distribution of Vim and GFAP during development and had a caudorostral sequence of appearance of immunoreactivities similar to that observed for ependymal and radial glia cells. Several circumventricular organs (the organon vasculosum hypothalami, saccus vasculosus, and area postrema) exhibited highly specialized Vim- and/or GFAP-expressing glial cells. The glial cells of the midline septa of several brain regions were also Vim and/or GFAP immunoreactive. In the adult brain, tanycytes retain Vim expression in several brain regions. As in other vertebrates, the regions with Vim-immunoreactive ventricular and midline glia may represent areas with the capability of plasticity and regeneration in adult brain.     

Filament proteins in rat optic nerves undergoing Wallerian degeneration: Localization of vimentin,the fibroblastic 100-Å filament protein,in normal and reactive astrocytes

A major polypeptide in the gliosed optic nerves of blinded rats was identified as vimentin, the fibroblastic, 100-Å filament protein typical of mesenchymal cells, by comigration experiments with purified vimentin on SDS-polyacrylamide gel electrophoresis. The localization of vimentin to astrocytes was confirmed by immunofluorescence microscopy with vimentin antisera. The distribution of vimentin and of astrocyte-specific glial fibrillary acidic (GFA) protein in cryostat sections of rat brain and spinal cord was different. Vimentin was confined to the main processes of fibrous astrocytes and could not be identified in the delicate framework of glial fibrils demonstrated with GFA antisera. Bergmann radial glia in the molecular layer of the cerebellum were exceptional in this respect, because they were equally well stained with vimentin and GFA antisera.     

GFAP-expressing cells in the postnatal subventricular zone display a unique glial phenotype intermediate between radial glia and astrocytes

Neural stem cells in the adult subventricular zone (SVZ) derive from radial glia and express the astroglial marker glial fibrillary acidic protein (GFAP). Thus, they have been termed astrocytes. However, it remains unknown whether these GFAP-expressing cells express the functional features common to astrocytes. Using immunostaining and patch clamp recordings in acute slices from transgenic mice expressing green fluorescent protein (GFP) driven by the promoter of human GFAP, we show that GFAP-expressing cells in the postnatal SVZ display typical glial properties shared by astrocytes and prenatal radial glia such as lack of action potentials, hyperpolarized resting potentials, gap junction coupling, connexin 43 expression, hemichannels, a passive current profile, and functional glutamate transporters. GFAP-expressing cells express both GLAST and GLT-1 glutamate transporters but lack AMPA-type glutamate receptors as reported for dye-coupled astrocytes. However, they lack 100 microM Ba2+-sensitive inwardly rectifying K+ (K(IR)) currents expressed by astrocytes, but display delayed rectifying K+ currents and 1 mM Ba2+-sensitive K+ currents. These currents contribute to K+ transport at rest and maintain hyperpolarized resting potentials. GFAP-expressing cells stained positive for both K(IR)2.1 and K(IR)4.1 channels, two major K(IR) channels in astrocytes. Ependymal cells, which also derive from radial glia and express GFAP, display typical glial properties and K(IR) currents consistent with their postmitotic nature. Our results suggest that GFAP-expressing cells in concert with ependymal cells can perform typical astrocytic functions such as K+ and glutamate buffering in the postnatal SVZ but display a unique set of functional characteristics intermediate between astrocytes and radial glia.     

Astroglial differentiation in the opossum superior colliculus

Glial markers, namely, vimentin, glial fibrillary acidic protein (GFAP), and glycogen, as well as accumulation of axon-borne horseradish peroxidase (HRP), were used to visualize radial glial cells in the developing opossum superior colliculus (SC) and to follow changes in young astrocytes of the superficial layers. Vimentin, GFAP, and glycogen are relatively abundant in elements of the median ventricular formation (MVF), which persists at least as late as weaning time, i.e., postconception day 103, postnatal day 90 (PND90). Radial profiles and end-feet in the remaining collicular sectors (main radial system, MRS) are also vimentin-positive but show little or no glycogen or anti-GFAP staining. The numeric density of MRS profiles is very high at the final stages of neuronal migration (PND12) but falls to vestigial numbers by PND 56-60. Antivimentin staining and filling of MRS profiles by axon-borne HRP disappear in parallel. Before total regression of MRS profiles, young astrocytes of the superficial gray layer exhibit a transiently high GFAP expression that is not found in those of the subjacent layers. The results suggest that 1) radial glia at or near the collicular midline are well equipped for a mechanical supportive role, and their abundant glycogen accumulation may reflect their eventual transformation in cells with high glycolytic metabolism, including tanycytes; 2) in most collicular sectors, some radial glia cells persist for long periods after cessation of neuronal migration and may interact with afferent fibers coursing through the superficial neuropil; 3) radially oriented astrocytes of the superficial gray layer exhibit a transiently high GFAP expression that is temporally correlated with late transformations of the retinocollicular projections.