Glial fibrillary acidic polypeptides in peripheral glia. Molecular weight, heterogeneity and distribution

Glial fibrillary acidic (GFA) polypeptides are present in major categories of rat peripheral glia including non-myelin-forming Schwann cells, enteric glia and some satellite cells. They can be detected both immunochemically and immunohistochemically. The immunoreactivity is associated with a polypeptide which has an MW of 49 000, indistinguishable from that of glial fibrillary acidic protein (GFAP) from rat brain. In spite of this, the GFA polypeptides found in the peripheral nervous system and central nervous system are not identical since they can be distinguished both immunohistochemically and immunochemically by a monoclonal GFAP antibody which recognizes GFAP in astrocytes and some enteric glia, but not GFAP in non-myelin-forming Schwann cells, satellite cells and many enteric glia. GFA-related molecules can also be detected in human Schwann cells by immunofluorescence. The results suggest, however, that the glial filament polypeptides of peripheral glia and astrocytes are less closely related in the human than in the rat. The glial distribution of GFAP is closely paralleled by 2 cell surface proteins, Ran-2 and A5E3 antigen. Although GFAP, Ran-2 and A5E3 are individually expressed by diverse cell types, the phenotype GFAP+, Ran-2+, A5E3+ defines a narrow group including only non-myelin-forming Schwann cells, enteric glia and astrocytes. These observations suggest that the non-myelin-forming cells of the central and peripheral nervous system may share some common functions.     

Enteric glia.

The structure of the enteric nervous system (ENS) is different from that of extraenteric peripheral nerve. Collagen is excluded from the enteric plexuses and support for neuronal elements is provided by astrocyte-like enteric glial cells. Enteric glia differ from Schwann cells in that they do not form basal laminae and they ensheath axons, not individually, but in groups. Although enteric glia are rich in the S-100 and glial fibrillary acidic proteins, it has been difficult to find a single chemical marker that distinguishes enteric glia from non-myelinating Schwann cells. Nevertheless, two monoclonal antibodies have been obtained that recognize antigens that are expressed on Schwann cells (Ran-1 in rats and SMP in avians) but not enteric glia. Functional differences between enteric glia and non-myelinating Schwann cells, including responses to gliotoxins and in vitro proliferative rates, have also been observed. Developmentally, enteric glia, like Schwann cells, are derived from the neural crest. In both mammals and birds the precursors of the ENS appear to migrate to the bowel from sacral as well as vagal levels of the crest. These crest-derived emigrés give rise to both enteric glia and neurons; however, analyses of the ontogeny of the enteric innervation in a mutant mouse (the ls/ls), in which the original colonizing waves of crest-derived precursor cells are unable to invade the terminal colon, suggest that enteric glia can also arise from Schwann cells that enter the gut with the extrinsic innervation. When induced to leave back-transplanted segments of avian bowel, enteric crest-derived cells migrate into peripheral nerves and form Schwann cells. Enteric glia and Schwann cells thus appear to be different cell types, but ones that derive from lineages that diverge relatively late in ontogeny.     

A cell surface protein of astrocytes, Ran-2, distinguishes non-myelin-forming Schwann cells from myelin-forming Schwann cells

Defining the molecular phenotype of adult glial cells in the peripheral nervous system in situ forms a good basis for subsequent studies on the development of these cells, and for determining the role of neurons in the attainment and maintenance of the mature glial phenotype. We report here the characterization of the glial surface antigen, Ran-2, and describe its distribution in the peripheral and central nervous system of adult rats. Immunoprecipitation of the antigen from cultured astrocytes with monoclonal Ran-2, antibodies, showed that Ran-2 is a protein with an apparent molecular weight of 140,000 daltons. In immunofluorescence studies of teased nerve preparations, Ran-2 was found on the surface of non-myelin-forming Schwann cells in all nerves surveyed, i.e. the sciatic, dorsal and ventral roots, cervical sympathetic trunk and the brachial plexus. In contrast, it was not detected immunohistochemically on myelin-forming Schwann cells. The antigen was also absent, or present at very low levels, on the satellite cells of dorsal root sensory ganglia, although, as we reported previously, it was present on the glial cells of the enteric nervous system. The Ran-2 antigen was also associated with perineurial cells. In short-term cell cultures of the sciatic nerve and cervical sympathetic trunk from 19-day-old rats, Ran-2 could be localized on the surface of individual viable Schwann cells. In the central nervous system, the antigen was present on astrocytes in sections of the optic nerve.     

Novel insights into the glia limitans of the olfactory nervous system

Olfactory ensheathing cells (OECs) are often described as being present in both the peripheral and the central nervous systems (PNS and CNS). Furthermore, the olfactory nervous system glia limitans (the glial layer defining the PNS–CNS border) is considered unique as it consists of intermingling OECs and astrocytes. In contrast, the glia limitans of the rest of the nervous system consists solely of astrocytes which create a distinct barrier to Schwann cells (peripheral glia). The ability of OECs to interact with astrocytes is one reason why OECs are believed to be superior to Schwann cells for transplantation therapies to treat CNS injuries. We have used transgenic reporter mice in which glial cells express DsRed fluorescent protein to study the cellular constituents of the glia limitans. We found that the glia limitans layer of the olfactory nervous system is morphologically similar to elsewhere in the nervous system, with a similar low degree of intermingling between peripheral glia and astrocytes. We found that the astrocytic layer of the olfactory bulb is a distinct barrier to bacterial infection, suggesting that this layer constitutes the PNS–CNS immunological barrier. We also found that OECs interact with astrocytes in a similar fashion as Schwann cells in vitro. When cultured in three dimensions, however, there were subtle differences between OECs and Schwann cells in their interactions with astrocytes. We therefore suggest that glial fibrillary acidic protein–reactive astrocyte layer of the olfactory bulb constitutes the glia limitans of the olfactory nervous system and that OECs are primarily “PNS glia.”     

Colonization of the bowel by the precursors of enteric glia: studies of normal and congenitally aganglionic mutant mice

The terminal portion of the ls/ls mouse is congenitally aganglionic because the precursors of enteric neurons fail to enter this region. This animal was studied in order to gain insight into the origin of enteric glia and into the process by which the precursors of these cells colonize the gut. In control (CD-1) mice, immunoreactivity of the glial marker, glial fibrillary acidic protein, appeared for the first time in the fetal bowel at day E16 and, in adults, was much more intense within intraenteric neural elements than in nerves outside the bowel. Glial fibrillary acidic protein developed in tissue cultures of fetal intestine explanted before the protein appeared in situ, and before the bowel became innervated by extrinsic nerves; thus, the precursors of cells able to elaborate glial fibrillary acidic protein must have been present, but unrecognizable, in the original explants. This explant assay demonstrated that these glial precursors were present in all regions of the bowel of control mice, but not in the presumptive aganglionic bowel of ls/ls mice. The nerves (of extrinsic origin) in the aganglionic tissue of ls/ls mice showed a high level of immunoreactive glial fibrillary acidic protein; nevertheless, their ultrastructure was typical of peripheral nerve, not enteric plexus, and they contained Schwann cells, not enteric glia. These observations support the view that enteric glia are derived from the single wave of neural crest colonists that populates the enteric nervous system before the gut receives its extrinsic innervation. These glial precursors, like neuronal precursors, tend to be excluded from the presumptive aganglionic ls/ls bowel. In contrast, Schwann cells grow into the abnormal ls/ls gut with the extrinsic innervation. The enteric microenvironment appears to promote the expression of glial fibrillary acidic protein in both enteric glia and Schwann cells; however, even within the bowel, Schwann cells retain their characteristic morphology. It is thus probable that the normal enteric nervous system contains supporting cells of separate lineages, enteric glia and Schwann cells.     

Isolation,identification, and culture of normal mouse colonic glia

We have developed a novel method of isolating and culturing murine colonic mucosal glial cells. Two morphologies are appreciated, a small flat bi or tri polar cell and a larger multipolar cell. The glial cultures have been freed of contaminating fibroblasts and epithelial cells and have been passaged by trypsinization. By intermediate filament (IF) typing, the glial cells have been further characterized as astrocyte-like. All cells expressed glial fibrillary acid protein but not neurofilament 160 protein. The glial cultures expressed the neuropeptides, substance P and substance K. Central nervous system astrocytes synthesize neuropeptides, prostaglandins and cytokines, and can express major histocompatibility class II antigens. It is likely that enteric mucosal glia will also prove to have varied functions. These cultures can now be used to define the role of enteric mucosal glia and to further study their complex interaction with other cells of the colonic mucosa.     

Developmental dynamics of peripheral glia in Drosophila melanogaster

To study the roles of peripheral glia in nervous system development, a thorough characterization of wild type glial development must first be performed. We present a developmental profile of peripheral glia in Drosophila melanogaster that includes glial genesis, developmental morphology, the establishment of transient cellular contacts, migration patterns, and the extent of nerve wrapping in the embryonic and larval stages. In early embryonic development, immature peripheral glia that are born in the CNS seem to be intermediate targets for neurites that are migrating into the periphery. During migration to the PNS, peripheral glia follow the routes of pioneer neurons. The glia preferentially adhere to sensory axonal projections, extending cytoplasmic processes along them such that by the end of embryogenesis peripheral glial coverage of the sensory system is complete. In contrast, significant lengths of motor branch termini are unsheathed in the mature embryo. During larval stages however, peripheral glia further extend and elaborate their cytoplasmic processes until they often reach to the neuromuscular junction. Throughout the embryonic and larval developmental stages, we have also observed a number of similarities of peripheral glia to vertebrate Schwann cells and astrocytes. Peripheral glia seem to have dynamic and diverse roles and their similarities to vertebrate glia suggest that Drosophila may serve as a powerful tool for analysis of glial roles in PNS development in the future.     

Expression of a functional metabotropic glutamate receptor 5 on enteric glia is altered in states of inflammation

The metabotropic glutamate receptor 5 (mGluR5) is expressed by astrocytes and its expression is modulated by inflammation. Enteric glia have many similarities to astrocytes and are the most numerous cell in the enteric nervous system (ENS). We investigated whether enteric glia express a functional mGluR5 and whether expression of this receptor was altered in colitis. In both enteric plexuses of the ileum and colon of guinea pigs and mice, we observed widespread glial mGluR5 expression. Incubation of isolated segments of the guinea pig ileum with the mGluR5 specific agonist RS-2-chloro-5-hydroxyphenylglycine (CHPG) caused a dose-dependent increase in the glial expression of c-Fos and the phosphorylated form of the extracellular signal-regulated kinase 1/2. Preincubation of tissues with the group I metabotropic glutamate receptor antagonist, S-4-carboxyphenylglycine, abolished the effects of CHPG. We examined mGluR5 expression in the guinea pig trinitrobenzene sulfonic acid and the IL-10 gene-deficient (IL-10(-/-)) mouse models of colitis. In guinea pigs, mGluR5 immunoreactivity became diffusely localized over the colonic myenteric ganglia, suggesting a change in receptor distribution. In contrast, glial mGluR5 expression was significantly reduced in the colonic myenteric plexus of IL-10(-/-) mice, as assessed with both real-time quantitative RT-PCR as well as immunohistochemistry and image analysis. These changes occurred without concomitant changes to enteric ganglia or glial fibrillary acidic protein expression in the IL-10(-/-) mouse. Our data suggest that enteric glia are a functional target of the glutamatergic neurotransmitter system in the ENS and that changes in mGluR5 expression may be of physiological significance during colitis.     

The olfactory nerve contains two populations of glia, identified both in vivo and in vitro.

The peripheral olfactory nervous system exhibits, uniquely, neuronal cell body replacement and reestablishment of central connections in adult mammals. The role of the olfactory nerve glia in these phenomena is unknown, but information might be provided by in vitro systems. This paper reports on the characterization of olfactory nerve glia in dissociated cell cultures of newborn rat nasal mucosal tissues. The predominant type of glial cell resembled Schwann cells and immunostained for the S-100 protein, found in all glial cell types; glial fibrillary acidic protein (GFAP), found in astrocytes and nonmyelinating Schwann cells; and showed binding of 217C, a monoclonal Schwann-cell marker that binds to the low-affinity NGF receptor in glioma cells. They were negative for A2B5. The Schwann-cell-like olfactory glia changed morphology upon culturing in serum-free medium, with further shape changes after plating on laminin. Plating on laminin increased cell numbers. A second population, found only after GFAP-immunostaining, was astrocyte-like in morphology and represented approximately 10 percent of all glial cells. These were S-100-, A2B5-, and 217C-negative, a unique glial cell immunological profile. At low dilutions of anti-GFAP (1/10,000), or with weak fluorescent secondary antibodies, astrocyte-like glia were immunostained but Schwann-cell-like glia were not detectable. Astrocyte-like glia were not an artifact of the dissection, since they were detectable in tissue sections of newborn-rat olfactory nerves immunostained with a low dilution of anti-GFAP. The presence of two types of glial cells in culture suggests similarities between olfactory glia and enteric glia.     

Analysis of enteric neurons, glia and their interactions using explant cultures of the myenteric plexus

The enteric nervous system (ENS) of the gastrointestinal tract is the largest and most complicated division of the peripheral nervous system. The ENS possesses reflex pathways composed of motor neurons, interneurons and sensory neurons which act in an integrated fashion together with input from the central nervous system to control gut function. The neurons, morphologically and electrophysiologically a very heterogeneous group containing a large number of different proven and putative neurotransmitters, are intimately associated with enteric glia, which both at the morphological and molecular level resemble astrocytes. In this review we describe how explant cultures from the ENS have been used to investigate the neurochemical, molecular and electrophysiological characteristics of ENS neurons, the molecular properties of enteric glia and their interactions with one another.     

Enteric glial cells: New players in Parkinson's disease?

Lewy pathology has been described in neurons of the enteric nervous system in nearly all Parkinson's disease (PD) patients at autopsy. The enteric nervous system not only contains a variety of functionally distinct enteric neurons but also harbors a prominent component of glial cells, the so‐called enteric glial cells, which, like astrocytes of the central nervous system, contribute to support, protect, and maintain the neural network. A growing body of evidence supports a role for enteric glial cells in the pathophysiology of gastrointestinal disorders such as inflammatory bowel disease and chronic constipation. We have recently shown that enteric glial cell dysfunction occurs in PD. In the present review, we discuss the possible implications of enteric glia in PD‐related gut dysfunction as well as in disease initiation and development. © 2014 International Parkinson and Movement Disorder Society     

Peripapillary glial cells in the chick retina: A special glial cell type expressing astrocyte, radial glia, neuron, and oligodendrocyte markers throughout development

Peripapillary glial cells of the chick are a special type of glia, not only because of their position, forming a boundary between the retina on one side and the optic nerve head (ONH) and the pecten on the other, but also because although they have the same orientation and similar shape as the retinal Müller cell (a type of radial glia) and express common markers for these cells and astrocytes, they do not express glutamine synthetase (GS) or carbonic anhydrase C (CA-C), enzymes intensely expressed by Müller cells and astrocytes. In this study, we present further molecular characterization of these cells, using immunohistochemistry techniques. We show that peripapillary glial cells express a novel neuron antigen, 3BA8, that in the adult retina is located only in one neuron type (the amacrine cell) and in the inner plexiform layer (IPL). They also express an antigen specific to myelin and oligodendrocytes, MOSP, and a glial antigen, 3CB2, expressed by radial glia and astrocytes throughout the CNS. The study of the developmental expression of these three antigens in the peripapillary glial cell territory shows different spatiotemporal labeling patterns: 3CB2 and 3BA8 are expressed much earlier (embryonic days E3 and E5, respectively) than MOSP (E12), and during a developmental window (E6-E10) 3BA8 labels the peripapillary glial cells intensely and does not label the ONH or the optic nerve (ON), which are labeled later. The expression of 3CB2 is much more intense in the peripapillary glial cells than in Müller cells from early stages of development up to E16, and the expression of MOSP starts earlier in the peripapillary glial cells than in the Müller cells and is maintained with much higher intensity in the peripapillary glial cells throughout development. These findings show that Müller and peripapillary glial cells follow independent courses of differentiation, which together with the fact that the peripapillary glial cells express molecules typical of neurons, oligodendrocytes, radial glia, and astrocytes are evidence that peripapillary glial cells are a unique type of glia in the CNS.     

Ran-2, a glial lineage marker,is a GPI-anchored form of ceruloplasmin

Cell interactions in the nervous system are frequently mediated by surface proteins that are attached to the membrane by a glycosyl phosphatidylinositol (GPI) anchor. In this study, we have characterized the expression of such proteins on glial cells. We have detected a major GPI-anchored protein on astrocytes and Schwann cells, with a molecular weight of 140 kD. When Schwann cells were treated with forskolin to promote a myelinating phenotype, expression of this 140-kD protein dramatically decreased, whereas another GPI-anchored protein of 80 kD was strongly induced; expression of other integral membrane proteins were likewise dramatically altered. The size and pattern of expression of the 140-kD protein suggested that it might correspond to the Ran-2 antigen, a glial lineage marker. This notion was confirmed by immunoprecipitating this 140-kD protein with the Ran-2 monoclonal antibody. The Ran-2 antigen is expressed over the entire Schwann cell surface in a punctate fashion; it is removed by phosphatidylinositol phospholipase C treatment, thereby confirming that it is GPI-anchored. When Schwann cells are cocultured with neurons, the Ran-2 antigen initially concentrates at sites of Schwann cell contact with neurons, suggesting that it may play a role in early Schwann cell–neuron interactions; it is then downregulated. Protein sequencing of the Ran-2 antigen immunopurified from rat brain membranes showed complete identity over two extended segments with the copper binding protein ceruloplasmin. These findings indicate that astrocytes and Schwann cells express a novel GPI-anchored form of ceruloplasmin and suggest that this GPI form plays a role in axonal–glial interactions. J. Neurosci. Res. 54:147–157, 1998. © 1998 Wiley-Liss, Inc.     

The Enteric Glia: Identity and Functions

Enteric glial cells were first described at the end of the 19th century, but they attracted more interest from researchers only in the last decades of the 20th. Although, they have a different embryological origin, the enteric GLIA share many characteristics with astrocytes, the main glial cell type of the central nervous system (CNS), such as in their expression of the same markers and in their functions. Here we review the construction of the enteric nervous system (ENS), with a focus on enteric glia, and also the main studies that have revealed the action of enteric glia in different aspects of gastrointestinal tract homeostasis, such as in the intestinal barrier, in communications with neurons, and in their action as progenitor cells. We also discuss recent discoveries about the roles of enteric glia in different disorders that affect the ENS, such as degenerative pathologies including Parkinson's and prion diseases, and in cases of intestinal diseases and injury. GLIA 2015;63:921–935     

A cell surface antigen expressed by astrocytes and their precursors

Increasing evidence suggests that astrocytes of the mammalian CNS are a heterogeneous population of cells that express a number of common characteristics. In most cases, astrocytes or their precursors contain a class of intermediate filaments, composed in large part of glial fibrillary acidic protein (GFAP). While the expression of GFAP immunoreactivity is a specific characteristic of astrocytes in the vertebrate CNS, not all astrocytes contain detectable levels of glial filaments, particularly during early development. We have isolated a monoclonal antibody termed 7B11 which binds to the surface of astrocytes and glial precursors, but not to other major types of neural cells. The 7B11 antigen is expressed by astrocytes in the adult CNS in vivo and in cultures of developing CNS tissue, but not on cells of the peripheral nervous system. During early development, 7B11 immunoreactivity appears prior to the expression of GFAP and is retained as punctate staining on the surface of most classes of astrocytes. During CNS maturation, however, 7B11 immunoreactivity is lost from the surface of Bergmann glia in the cerebellum, suggesting that differentiative events lead to functionally and anti-genically distinct classes of CNS glial cells. In the adult spinal cord, biochemical analysis suggests that the epitope recognized by 7B11 is associated with a group of polypeptides of apparent molecular weights 200-160, 140, and 92 kD. The cellular distribution of 7B11 expression suggests that astrocytes and their precursors share a distinct cell surface antigenic property and that the expression of 7B11 immunoreactivity may be a useful operational marker for astrocytes in the absence of detectable GFAP expression. © 1993 Wiley-Liss, Inc.     

Neuroligin 3 is a vertebrate gliotactin expressed in the olfactory ensheathing glia, a growth-promoting class of macroglia

The molecular mechanisms that drive glia-glial interactions and glia-neuronal interactions during the development of the nervous system are poorly understood. A number of membrane-bound cell adhesion molecules have been shown to play a role, although the precise nature of their involvement is unknown. One class of molecules with cell adhesive properties used in the nervous system is the serine-esterase-like family of transmembrane proteins. A member of this class, a glia-specific protein called gliotactin, has been shown to be necessary for the development of the glial sheath in the peripheral nervous system of Drosophila melanogaster. Gliotactin is essential for the development of septate junctions in the glial sheath of individual and neighboring glia. Mutations that remove this protein result in paralysis and eventually death due to a breakdown in the glial-based blood-nerve barrier. To study the role of gliotactin during vertebrate nervous system development, we have isolated a potential vertebrate gliotactin homologue from mice and rat and found that it corresponds to neuroligin 3. Using a combination of RT-PCR and immunohistochemistry, we have found that neuroligin 3 is expressed during the development of the nervous system in many classes of glia. In particular neuroligin 3 is expressed in the olfactory ensheathing glia, retinal astrocytes, Schwann cells, and spinal cord astrocytes in the developing embryo. This expression is developmentally controlled such that in postnatal and adult stages, neuroligin 3 continues to be expressed at high levels in the olfactory ensheathing glia, a highly plastic class of glia that retain many of their developmental characteristics throughout life.     

Astrocytes and Bergmann glia as an important site of nitric oxide synthase I

In the central nervous system nitric oxide appears to be critically involved in a number of physiological and pathological processes. Although there is convincing evidence for expression of nitric oxide synthase in cultured glial cells, demonstration of this enzyme in glial cells in situ remained largely unsatisfactory. In the present study we applied immunostaining to freeze-dried sections of snap-frozen hippocampi and cerebella of rats and to sections of freeze-dried brain tissue in order to minimize diffusion artefacts and thus to obtain more precise information about the true in situ localization of nitric oxide synthase. Here we show that astrocytes and Bergmann glia react strongly with antibodies raised against cerebellar nitric oxide synthase and against a type I nitric oxide synthase-specific C-terminal peptide, respectively. This finding was further substantiated by histochemical localization of NADPH-diaphorase activity in astrocytes and Bergmann glia as well as by immunoreactivity of both types of glia cells with antibodies to the NADPH-delivering enzyme glucose-6-phosphate dehydrogenase. We conclude, that astrocytes are important sites of nitric oxide synthase I in brain, suggesting that these cells might use nitric oxide as gaseous messenger molecule for various aspects of glia-neuron signalling. © 1996 Wiley-Liss, Inc.     

Morphology and differentiation of radial glia in the developing rat spinal cord

Important events underlying the proper functioning of the central nervous system (CNS) include the production, assembly, and differentiation of appropriate types and numbers of cells during development. The mechanisms that control these events are difficult to unravel because of displacement of cells from their sites of origin to their permanent locations and because of the diverse cellular composition of the CNS. As in other regions of the mammalian CNS, the two major classes of neuroglial cells in the rat spinal cord are oligodendrocytes and astrocytes. In the developing spinal cord, radial glia are prominent. In this study, radial glia in the cervical region of the spinal cord were analysed. 1,1'Dioctadecyl-3,3,3'-tetramethylindocarbocyanine perchlorate (DiI) was used to determine the morphology and distribution of radial glia during spinal cord development. The DiI labelling technique enabled locating glial precursor cells during spinal cord development. Radial fibres that extended from the central canal to the pial surface were present at embryonic days 14, 16, and 18 in the developing spinal cord. Their distribution was restricted with increasing development, and by embryonic day 20 the only remaining evidence of radial glia were short radial processes in the white matter.     

Properties of human astrocytes and NG2 glia

Since animal models are inevitable for medical research, information on species differences in glial cell properties is critical for successful translational research. Here, we review current knowledge about morphological and functional properties of human astrocytes and NG2 glial cells and compare these data with those obtained for the comparable cells in rodents. Morphological analyses of astrocytes in the neocortex of rodents versus humans have demonstrated clear differences. In contrast, the functional properties of astrocytes or NG2 glial cells in these species are surprisingly similar. However, these findings should be interpreted with caution, as so far functional analyses of human cells are only available from neocortex and hippocampus, and it is known from rodent studies that the properties of astrocytes in different brain regions may vary considerably. Moreover, technical challenges render astrocyte electrophysiological measurements in situ unreliable, and human cell properties may be affected by medications. Nevertheless, based on the limited data currently available, there is substantial similarity between human and rodent astrocytes with regard to those functional properties studied to date. The unique morphological characteristics of astrocytes in human neocortex call for further physiological analysis. The basic properties for NG2 glia are even less completely evaluated with regard to the question of species differences but no glaring differences have been reported so far. In conclusion, it remains justifiable to employ mouse or rat models to investigate the etiology of human CNS diseases that might involve astrocytes or NG2 glia.