New insights on the biology of myelin basic protein gene: the neural-immune connection

In the past 6 years, our conception of the major myelin protein genes has begun to change significantly because of recent findings documenting the existence of new exons encoding other products of these genes. A decade ago the myelin basic protein (MBP) and proteolipid protein (PLP) genes were thought to be expressed solely in myelin-forming cells, and their products were thought to be structural components of myelin. Since then, abundant evidence has been gathered identifying the presence of products of these genes in nonmyelinating cell types including both the immune and the nervous systems. Furthermore, within the nervous system, products of these genes have been identified in neurons and embryonic cells, clearly indicating that these myelin protein genes have additional functions in a number of cell types that are unrelated to myelination. In this brief communication, we review the recent literature that has resulted in this revision of our understanding of the MBP gene structure, products and expression.     

Repair of a myelin lesion by Schwann cells transplanted in the adult mouse spinal cord

In multiple sclerosis and experimental demyelination, oligodendrocytes and Schwann cells are able to repair myelin lesions of the central nervous system. However, spontaneous myelin repair is often insufficient. Several approaches to enhance remyelination have been considered and transplantation of myelin-forming cells has been proposed as one of them. In this paper, we present results which confirm the ability of transplanted Schwann cells to remyelinate an induced lesion of the spinal cord. Schwann cells were either purified Schwann cells isolated from 1–day-old rat sciatic nerves, or immortalized Schwann cells (MSC80) arising from a purified culture of 7-day-old mouse sciatic nerves. They were transplanted into or at a distance from a lysolecithin-induced lesion of the Shiverer spinal cord. Labelling of the Schwann cells with the fluorochrome Hoechst 33342 enabled us to trace them after transplantation in their host and evaluate their ability to reach and to repair the demyelinated lesion. Using the Hoechst-Shiverer model, we show that when transplanted in the lesion, cultured Schwann cells, even immortalized, are able to remyelinate such a lesion efficiently. In addition, when transplanted at a distance from the lesion, they are able to reach and repair the lesion in time frames which allow them to complete actively with host oligodendrocytes.     

Remyelination Therapy for Multiple Sclerosis

Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system characterized by infiltration of immune cells and progressive damage to myelin and axons. All therapeutics used to treat MS have been developed to target an overactive immune response, with aims to reduce disease activity. Chronic demyelinated axons are further prone to irreversible damage and death, and it is imperative that new therapies address this critical issue. Remyelination, the generation of new myelin in the adult nervous system, is an endogenous repair mechanism that restores function of denuded axons and delays their deterioration. Although remyelination can be extensive in some patients, the majority of cases limit repair only to the acute phase of disease. A significant current drive in new MS therapeutics is to identify targets that can promote remyelination by boosting endogenous oligodendrocyte precursor cells to form new myelin. Also, a number of inhibitory pathways have been identified in chronic MS lesions that prevent oligodendrocyte precursor cells from being properly recruited to demyelinated lesions or interfere with their differentiation to myelin-forming oligodendrocytes. In this review, we introduce the phenomenon of remyelination from the view of experimental models and studies in MS patients, describe a potential role in remyelination for currently available MS mediations, and discuss many avenues that are being actively studied to promote remyelination. The next frontier in MS therapeutics will supplement immunomodulation with agents that directly foster myelin repair, with aims to delay disease progression and recover lost neurological functions.     

Distribution of adrenocorticoid receptors in the rat CNS measured by competitive PCR and cytosolic binding

A growing number of glycoproteins have been identified and characterized in myelin and myelin-forming cells. In addition to the major P0 glycoprotein of compact PNS myelin and the myelin-associated glycoprotein (MAG) in the periaxonal membranes of myelin-forming oligodendrocytes and Schwann cells, the list now includes peripheral myelin protein-22 (PMP-22), a 170 kDa glycoprotein associated with PNS myelin and Schwann cells (P170k/SAG), Schwann cell myelin protein (SMP), myelin/oligodendrocyte glycoprotein (MOG), and oligodendrocytemyelin glycoprotein (OMgp). Many of these glycoproteins are members of the immunoglobulin superfamily and express the adhesion-related HNK-1 carbohydrate epitope. This review summarizes recent findings concerning the structure and function of these glycoproteins of myelin sheaths with emphasis on the physiological roles of oligosaccharide moieties.     

Features and Functions of Oligodendrocytes and Myelin Proteins of Lower Vertebrate Species

The myelin-forming cells in the central nervous system (CNS) of lower vertebrate species, in particular those of fish, profoundly differ from their mammalian counterparts in their biochemical phenotype in that they express Po-like glycoproteins as major myelin protein constituents instead of proteolipid protein, while in their overall cellular structure and their cell lineage relationships, they closely resemble mammalian oligodendrocytes. While molecular biology in the past has allowed to appropriately classify the major myelin proteins synthesized by fish oligodendrocytes, heterologous expression studies are expected to give a deeper insight into the particular features and the conserved functions of these proteins required for myelin formation and maintenance in fish. It is hoped that this approach will also help to improve our understanding of the molecular processes underlying the unique capacity of fish oligodendrocytes for remyelination after injury in the CNS. This survey may stimulate neuroscientists to engage into this exciting field.     

Modification of Schwann cell phenotype with Plp transgenes: Evidence that the PLP and DM20 isoproteins are targeted to different cellular domains

The X-linked proteolipid protein (Plp) gene encodes PLP, the major protein of central nervous system myelin, and its alternative RNA splice product, termed DM20. Schwann cells also express the Plp gene but, in contrast to oligodendrocytes, neither protein is incorporated into peripheral myelin. In the present study, we use different transgenes encoding PLP and DM20 to modify the expression of these proteins in myelin-forming Schwann cells of wild-type and jimpy mice. Increasing the level of PLP, either singly or in combination with DM20, leads to the incorporation of PLP into the compacted myelin sheath; however, DM20 always remains restricted to cytoplasmic regions of the Schwann cell. The insertion of PLP into the membrane does not appear to depend on a cooperativity of the two isoproteins. The presence of PLP does not visibly alter the ultrastructure and periodicity of peripheral nervous system (PNS) myelin. The results indicate that the absence of PLP in the peripheral myelin of normal animals most probably reflects the very low amounts of this isoprotein synthesised by Schwann cells. The preferential incorporation of PLP, as opposed to DM20, in peripheral myelin may indicate that a myelin targeting signal is present in the PLP-specific region of the molecule. J. Neurosci. Res. 50:13–22, 1997. © 1997 Wiley-Liss, Inc.     

Oligodendrogenesis in the subventricular zone and the role of epidermal growth factor

Demyelinating diseases are characterized by an extensive loss of oligodendrocytes and myelin sheaths from axolemma. These neurological disorders are a common cause of disability in young adults, but so far, there is no effective treatment against them. It has been suggested that neural stem cells (NSCs) may play an important role in brain repair therapies. NSCs in the adult subventricular zone (SVZ), also known as Type-B cells, are multipotential cells that can self-renew and give rise to neurons and glia. Recent findings have shown that cells derived from SVZ Type-B cells actively respond to epidermal-growth-factor (EGF) stimulation becoming highly migratory and proliferative. Interestingly, a subpopulation of these EGF-activated cells expresses markers of oligodendrocyte precursor cells (OPCs). When EGF administration is removed, SVZ-derived OPCs differentiate into myelinating and pre-myelinating oligodendrocytes in the white matter tracts of corpus callosum, fimbria fornix and striatum. In the presence of a demyelinating lesion, OPCs derived from EGF-stimulated SVZ progenitors contribute to myelin repair. Given their high migratory potential and their ability to differentiate into myelin-forming cells, SVZ NSCs represent an important endogenous source of OPCs for preserving the oligodendrocyte population in the white matter and for the repair of demyelinating injuries.     

Remyelination after olfactory ensheathing cell transplantation into diverse demyelinating environments

Olfactory ensheathing cells (OECs) can remyelinate demyelinated spinal cord axons when transplanted into chemically induced demyelinated lesions. Cell transplantation is typically performed within a few days after lesion induction, i.e. during active demyelination when myelin debris, cytokine level increases and macrophage/microglia activation is extensive. Inflammatory signaling has been suggested to facilitate remyelination in cell transplant studies. In this review we discuss the migration and remyelination properties of OECs transplanted into various demyelinating lesion environments including conditions when inflammation is active and when it is largely subsided. While sharing many common properties, comparisons of the in vivo fate between OECs and SCs suggest unique properties of OECs as compared to SCs. A complicating factor in the assessment of experimental remyelination by transplantation of myelin-forming cells in general is the rapidity of endogenous myelin repair in most rodent models of demyelination. Alternative persistent demyelination models are discussed as potential tools to study both the competency of chronic demyelinated axons for remyelination and the remyelination potential of cells such as human progenitors that require longer times to mobilize and remyelinate axons. This article is part of a Special Issue entitled: Understanding olfactory ensheathing glia and their prospect for nervous system repair.     

Generation and biological properties of a monoclonal antibody to Galactocerebroside

Galactocerebroside ( GalC ) is a major glycolipid of myelin and myelin-forming cells. We have generated a mouse IgM monoclonal antibody to GalC (M-anti- GalC ) which bound only to oligodendrocytes in rat and bovine central nervous system cultures as assessed by immunofluorescence. Double staining with rabbit anti-glial fibrillary acidic protein and anti-fibronectin antisera revealed no binding of M-anti- GalC to astrocytes or fibroblasts. Schwann cells, but not fibroblasts, were stained in short-term cultures of rat Schwann cells. M-anti- GalC exhibited in vitro cytotoxicity to rat and bovine oligodendrocytes in the presence of complement. This monoclonal antibody with its monospecificity, consistent titer, and capacity to induce cell lysis should be useful for in vitro and in vivo investigations concerning myelination and demyelination.     

Intraventricular transplantation of oligodendrocyte progenitors into a fetal myelin mutant results in widespread formation of myelin.

Transplantation of myelin-forming cells is a promising strategy for the treatment of myelin disorders. In this study, transplantation of glial cell progenitors into the cerebral ventricles of the embryonic myelin-deficient rat, a model of Pelizaeus-Merzbacher disease, was performed to assess the ability of these cells to incorporate into the developing brain and produce myelin. The donor cells migrated into the white and gray matter and produced myelin at widespread sites ranging from the corpus callosum and optic nerve to the cerebellum. These data suggest that myelin repair might be achieved by intraventricular delivery and transependymal incorporation of myelin-producing cells. Because these cells were genetically transduced to express a reporter gene, similar ex vivo manipulation with genes known to promote survival, migration, or proliferation of the transplanted cells could be used to enhance repair. Such a therapeutic strategy may be feasible in patients with inherited myelin disorders or in multiple sclerosis, particularly where the lesions are periventricular.     

Coordinated control of oligodendrocyte development by extrinsic and intrinsic signaling cues

Oligodendrocytes, the myelin-forming cells for axon ensheathment in the central nervous system, are critical for maximizing and maintaining the conduction velocity of nerve impulses and proper brain function. Demyelination caused by injury or disease together with failure of myelin regeneration disrupts the rapid propagation of action potentials along nerve fibers, and is associated with acquired and inherited disorders, including devastating multiple sclerosis and leukodystrophies. The molecular mechanisms of oligodendrocyte myelination and remyelination remain poorly understood. Recently, a series of signaling pathways including Shh, Notch, BMP and Wnt signaling and their intracellular effectors such as Olig1/2, Hes1/5, Smads and TCFs, have been shown to play important roles in regulating oligodendrocyte development and myelination. In this review, we summarize our recent understanding of how these signaling pathways modulate the progression of oligodendrocyte specification and differentiation in a spatiotemporally-specific manner. A better understanding of the complex but coordinated function of extracellular signals and intracellular determinants during oligodendrocyte development will help to devise effective strategies to promote myelin repair for patients with demyelinating diseases.     

Demyelination in the central nervous system mediated by an anti-oligodendrocyte antibody

The factors responsible for the major demyelinating disease of the central nervous system (CNS), multiple sclerosis, are poorly defined. Although T-cell-mediated immune responses play a pivotal role in establishing the inflammatory response, humoral factors also may be critical in disease progress. We have isolated a mouse monoclonal antibody (mAb 2B10) that recognizes a cell-surface molecule expressed exclusively by rat oligodendrocytes, the cells responsible for the formation and maintenance of CNS myelin. In cultures of neonatal rat spinal cord, mAb 2B10 specifically mediated oligodendrocyte cell death in the absence of complement. In the current study, mAb 2B10–producing hybridoma cells were implanted into adult rat brain ventricles, and the effect of mAb 2B10 on CNS cytoarchitecture was examined. In the optic nerves of mAb 2B10–treated animals, there was significant focal myelin degeneration near the optic chiasm. Axons in the myelin degenerate regions were largely healthy. There was no significant infiltration of hematopoietic-derived cells into the affected regions, but microglia were activated focally and phagocytosed the collapsed myelin. This study demonstrates that an antibody directed against myelin-forming cells induces CNS demyelination and supports the hypothesis that autoantibodies may play a role in CNS demyelinating diseases. J. Neurosci. Res. 54:158–168, 1998. © 1998 Wiley-Liss, Inc.     

Fibroblast growth factor-2 (FGF-2) and platelet-derived growth factor AB (PDGF AB) promote adult SVZ-derived oligodendrogenesis in vivo

The capacity of multipotential progenitor cells of the adult mammalian forebrain to generate myelin-forming oligodendrocytes was tested by grafting fragments of different regions of the subventricular zone (SVZ) of the lateral ventricle and the striatum of 6-month-old wild-type mice into the brain of neonate shiverer and wild-type mice. Without growth factor treatment, only few cells of the rostral SVZ survived and formed myelin after engraftment. Treating donors prior to transplantation with a single intraperitoneal injection of epidermal growth factor, basic fibroblast growth factor 2 (FGF-2), and platelet-derived growth factor AB (PDGF(AB)) vigorously promoted the survival, migration, and differentiation of the grafted SVZ cells into myelin-forming oligodendrocytes. In situ, both growth factors expanded the constitutively proliferative PSA-NCAM+ population and favored their differentiation toward the neuronal and oligodendroglial cell fate. The adult central nervous system thus harbors a focal reservoir of FGF-2 and PDGF(AB)-responsive cells which are able to generate substantial amounts of myelin-forming oligodendrocytes in vivo, opening a new prospective area for therapy in demyelinating diseases.     

Schwann cell expression of an oligodendrocyte-like remyelinating pattern after ethidium bromide injection in the rat spinal cord

Schwann cells are recognized by their capacity of producing single internodes of myelin around axons of the peripheral nervous system. In the ethidium bromide (EB) model of primary demyelination in the brainstem, it is observed the entry of Schwann cells into the central nervous system in order to contribute to the myelin repair performed by the oligodendrocytes that survived to the EB gliotoxic action, being able to even remyelinate more than one axon at the same time, in a pattern of repair similar to the oligodendroglial one. The present study was developed in the spinal cord to observe if Schwann cells maintained this competence of attending simultaneously different internodes. It was noted that, on the contrary of the brainstem, Schwann cells were the most important myelinogenic cells in the demyelinated site and, although rare, also presented the capacity of producing more than one internode of myelin in distinct axons.     

Olfactory ensheathing cells exhibit unique migratory, phagocytic, and myelinating properties in the X-irradiated spinal cord not shared by Schwann cells

Although several studies have shown that Schwann cells (SCs) and olfactory ensheathing cells (OECs) interact differently with central nervous system (CNS) cells in vitro, all classes of adult myelin-forming cells show poor survival and migration after transplantation into normal CNS. X-irradiation of the spinal cord, however, selectively facilitates migration of oligodendrocyte progenitor cells (OPCs), but not SCs, revealing differences in in vivo migratory capabilities that are not apparent in intact tissue. To compare the in vivo migratory properties of OECs and SCs and evaluate the potential of migrating cells to participate in subsequent repair, we first transplanted freshly isolated GFP-expressing adult rat olfactory bulb-derived OECs and SCs into normal and X-irradiated spinal cords. Both OECs and SCs showed limited survival and migration in normal spinal cord at 3 weeks. However, OECs, unlike SCs, migrated extensively in both grey and white matter of the X-irradiated spinal cord, and exhibited a phagocytic phenotype with OX-42 staining on their processes. If a X-irradiated and OEC transplanted spinal cord was then subjected to a focal demyelinating lesion 3 weeks after transplantation, OECs moved into the delayed demyelinated lesion and remyelinated host axons with a peripheral-like pattern of myelin. These results revealed a clear difference between the migratory properties of OECs and SCs in the X-irradiated spinal cord and demonstrated that engrafted OECs can participate in repair of subsequent lesions.     

Remyelination of the spinal cord following intravenous delivery of bone marrow cells

Bone marrow contains a population of pluripotent cells that can differentiate into a variety of cell lineages, including neural cells. When injected directly into the demyelinated spinal cord they can elicit remyelination. Recent work has shown that following systemic delivery of bone marrow cells functional improvement occurs in contusive spinal cord injury and stroke models in rat. We report here that secondary to intravenous introduction of an acutely isolated bone marrow cell fraction (mononuclear fraction) from adult rat femoral bones separated on a density gradient, ultrastructurally defined remyelination occurs throughout a focal demyelinated spinal cord lesion. The anatomical pattern of remyelination was characteristic of both oligodendrocyte and Schwann cell myelination; conduction velocity improved in the remyelinated axons. When the injected bone marrow cells were transfected to express LacZ, beta-galactosidase reaction product was observed in some myelin-forming cells in the spinal cord. Intravenous injection of other myelin-forming cells (Schwann cells and olfactory ensheathing cells) or the residual cell fraction of the gradient did not result in remyelination, suggesting that remyelination was specific to the delivery of the mononuclear fraction. While the precise mechanism of the repair, myelination by the bone marrow cells or facilitation of an endogenous repair process, cannot be fully determined, the results demonstrate an unprecedented level of myelin repair by systemic delivery of the mononuclear cells.     

Shark CNS myelin contains four polypeptides related to the PNS protein Po of higher classes

The relationship between the proteins of shark central nervous system (CNS) myelin and those of myelin from higher classes has been investigated using antibodies raised against a 31,500 molecular weight polypeptide from shark myelin. The antibodies cross-reacted with 3 shark CNS polypeptides apart from the original antigen, with 2 major polypeptides from shark peripheral nervous system myelin, with the Po protein from chicken and sheep peripheral nervous system myelin, but with none of the components of bovine CNS myelin. It appears that the oligodendroglial cells of the shark synthesize a protein closely related to the Po protein produced by Schwann cells of vertebrate classes above and including chondrichthytes.     

Transplantation of an acutely isolated bone marrow fraction repairs demyelinated adult rat spinal cord axons.

The potential of bone marrow cells to differentiate into myelin-forming cells and to repair the demyelinated rat spinal cord in vivo was studied using cell transplantation techniques. The dorsal funiculus of the spinal cord was demyelinated by x-irradiation treatment, followed by microinjection of ethidium bromide. Suspensions of a bone marrow cell fraction acutely isolated from femoral bones in LacZ transgenic mice were prepared by centrifugation on a density gradient (Ficoll-Paque) to remove erythrocytes, platelets, and debris. The isolated cell fraction contained hematopoietic and nonhematopoietic stem and precursor cells and lymphocytes. The cells were transplanted into the demyelinated dorsal column lesions of immunosuppressed rats. An intense blue beta-galactosidase reaction was observed in the transplantation zone. The genetically labeled bone marrow cells remyelinated the spinal cord with predominately a peripheral pattern of myelination reminiscent of Schwann cell myelination. Transplantation of CD34(+) hematopoietic stem cells survived in the lesion, but did not form myelin. These results indicate that bone marrow cells can differentiate in vivo into myelin-forming cells and repair demyelinated CNS.     

Cell-based remyelinating therapies in multiple sclerosis: evidence from experimental studies

PURPOSE OF REVIEW: Spontaneous remyelination occurs in the central nervous system of patients with multiple sclerosis. However, this process is not robust enough to promote a functional and stable recovery of the myelin architecture. The development of cell-based therapies, aimed at promoting multifocal remyelination, is therefore foreseen. RECENT FINDINGS: Several experimental cell-based strategies aimed at replacing damaged myelin-forming cells have been developed in the last few years. However, most of these therapeutic approaches - although consistently able to form new myelin sheaths at the transplantation site - are unfeasible owing to the mutifocality of the demyelinating process in multiple sclerosis patients and the inability to grow and produce large numbers of differentiated myelin-forming cells in vitro. Stem cell-based therapies that partially overcome these limitations have been proposed recently. SUMMARY: Stem cell-based remyelinating therapies can be considered a plausible alternative strategy in immune-mediated demyelinating disorders. However, before any potential applications in patients with multiple sclerosis can be envisaged, it is necessary to confront the following preliminary, and still unsolved, questions: (1) the ideal stem cell source for transplantation; (2) the most appropriate route of stem cell administration; and, last but not least, (3) the best approach for achieving an appropriate, functional and long-lasting integration of transplanted stem cells into the host tissue.     

Adaptive plasticity of Xenopus glial cells in vitro and after CNS fiber tract lesions in vivo

Xenopus oligodendrocytes and aspects of their differentiation were analyzed in vitro and in vivo using cell- and stage-specific antibodies. Undifferentiated oligodendrocytes were derived from optic nerves or spinal cords. They divided in vitro, were of elongated shape, were glial fibrillary acidic protein and O4 positive, transiently exhibited several antigens including HNK-1 and L1, and promoted axon growth as do Schwann cells. With forskolin they differentiated and, much like myelin-forming oligodendrocytes in the intact optic nerve and spinal cord, they expressed sets of advanced myelin markers. These advanced myelin markers disappeared from the regenerating optic nerve 4 weeks after lesion. The optic nerve instead was populated by cells with radial processes and somata in the center of the nerve; among them were cells and processes that were O4 positive and that are suspected to represent undifferentiated oligodendrocytes. Where processes of these cells reached to the retinal axons in the nerve's periphery, advanced myelin markers typical of differentiated oligodendrocytes reappeared 8 weeks after lesion. These glial changes did not occur in the absence of retinal axons. Thus, the apparent capability of Xenopus oligodendrocytes to adapt to the transient absence, reappearance, and regenerative state of the axons enables them to contribute to central nervous system fiber tract repair. This occurs in the lesioned optic nerve but not in the spinal cord, where no such glial changes were observed and where axons fail to regenerate. © 1996 Wiley-Liss, Inc.