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Biomedical subjects

A Ramón-Cueto

Publications and source records attributed to A Ramón-Cueto.

12 recordsLinked to original sources
PubMed2001

Cell therapy to repair injured spinal cords: olfactory ensheathing glia transplantation.

The absence of spontaneous axonal regeneration in the adult mammalian central nervous system cause devastating functional consequences in patients with spinal cord injuries. During the past decades several attempts have been made in order to find a strategy to repair injured spinal cords in experimental animals, that could provide a novel therapeutic approach in humans. Cell transplantation has been broadly used as an intervention to influence neuronal survival and axonal regeneration in the severed neuraxis. Of the cell types used for transplantation, olfactory ensheathing glia (OEG) promoted a dramatic functional improvement and anatomical repair after complete transection of the adult mammalian spinal cord. These cells can be easily obtained from adult donors opening the possibility of autologous transplantation. Grafting OEG to repair injured spinal cords offers some advantages compared to injections of other cell types. Therefore, OEG have become good candidates to bring about repair in damaged spinal cords. In this article we review OEG transplantation studies, discuss the properties that could account for their axonal growth-promoting ability, and the advantages of using OEG as a repair strategy.

Animals↗
PubMed2000

Functional recovery of paraplegic rats and motor axon regeneration in their spinal cords by olfactory ensheathing glia.

Axonal regeneration in the lesioned mammalian central nervous system is abortive, and this causes permanent disabilities in individuals with spinal cord injuries. In adult rats, olfactory ensheathing glia (OEG) transplants successfully led to functional and structural recovery after complete spinal cord transection. From 3 to 7 months post surgery, all OEG-transplanted animals recovered locomotor functions and sensorimotor reflexes. They presented voluntary hindlimb movements, they supported their body weight, and their hindlimbs responded to light skin contact and proprioceptive stimuli. In addition, relevant motor axons (corticospinal, raphespinal, and coeruleospinal) regenerated for long distances within caudal cord stumps. Therefore, OEG transplantation provides a useful repair strategy in adult mammals with traumatic spinal cord injuries. Our results with these cells could lead to new therapies for the treatment of spinal cord lesions in humans.

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PubMed1999

Two modes of microtubule-associated protein 1B phosphorylation are differentially regulated during peripheral nerve regeneration.

Two major modes of MAP1B phosphorylation (I and II), respectively recognized by monoclonal antibodies 150 and 125, have been related to remodeling and formation of processes in the mature nervous system. To gain insight into the cytoskeletal modifications underlying peripheral nerve regeneration, the pattern of expression of both MAP1B phosphorylated modes was studied during this process. Sciatic nerves from adult Wistar rats were crushed and animals allowed to survive for 5, 7, 10 or 14 days. After those survival periods, damaged and undamaged sciatic nerves, dorsal root ganglia (DRG), and spinal cords, were subjected to immunohistochemistry and Western blot, using antibodies 150 and 125. At all survival periods analysed, MAP1B phosphorylated at mode I was concentrated at the distal region of regenerating nerves whereas mode II phosphorylation underwent an overall decrease in regenerating axons that was less evident in more proximal nerve regions. Very high levels of MAP1B phosphorylated at mode II were detected in the bodies of DRG neurons and in bodies and dendrites of spinal motor neurons. This phosphorylation mode was also encountered in some Schwann cells and oligodendroglia associated with more proximal regions of regenerating axons. In this study we conclude that MAP1B was differentially phosphorylated depending on the cell type, subcellular compartment and stage of the regenerative process and discuss the possible functional implications that differential expression of each MAP1B phosphorylation mode might have during nerve regeneration.

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PubMed1998

Long-distance axonal regeneration in the transected adult rat spinal cord is promoted by olfactory ensheathing glia transplants.

The lack of axonal regeneration in the injured adult mammalian spinal cord leads to permanent functional impairment. To induce axonal regeneration in the transected adult rat spinal cord, we have used the axonal growth-promoting properties of adult olfactory bulb ensheathing glia (EG). Schwann cell (SC)-filled guidance channels were grafted to bridge both cord stumps, and suspensions of pure (98%) Hoechst-labeled EG were stereotaxically injected into the midline of both stumps, 1 mm from the edges of the channel. In EG-transplanted animals, numerous neurofilament-, GAP-43-, anti-calcitonin gene-related peptide (CGRP)-, and serotonin-immunoreactive fibers traversed the glial scars formed at both cord-graft interfaces. Supraspinal serotonergic axons crossed the transection gap through connective tissue bridges formed on the exterior of the channels, avoiding the channel interior. Strikingly, after crossing the distal glial scar, these fibers elongated in white and periaqueductal gray matter, reaching the farthest distance analyzed (1.5 cm). Tracer-labeled axons present in SC grafts were found to extend across the distal interface and up to 800 microm beyond in the distal cord. Long-distance regeneration (at least 2.5 cm) of injured ascending propriospinal axons was observed in the rostral spinal cord. Transplanted EG migrated longitudinally and laterally from the injection sites, reaching the farthest distance analyzed (1.5 cm). They moved through white matter tracts, gray matter, and glial scars, overcoming the inhibitory nature of the CNS environment, and invaded SC and connective tissue bridges and the dorsal and ventral roots adjacent to the transection site. Transplanted EG and regenerating axons were found in the same locations. Because EG seem to provide injured spinal axons with appropriate factors for long-distance elongation, these cells offer new possibilities for treatment of CNS conditions that require axonal regeneration.

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PubMed1998

Two novel monoclonal antibodies (1.9.E and 4.11.C) against olfactory bulb ensheathing glia.

We produced and characterized two monoclonal antibodies, termed 1.9.E and 4.11.C, that specifically recognize olfactory bulb ensheathing glia. Both antibodies were generated using the olfactory nerve layer (ONL) of newborn rat olfactory bulbs (P0, P1) as immunogens. The specificity of these antibodies was tested by immunofluorescence techniques on tissue sections and cultures of adult and neonatal rat olfactory bulbs, and by Western blot analysis. 1.9.E labeled the ONL and glomerular layer of the olfactory bulb (OB) of adult rats. In newborn rats, 1.9.E immunostained ensheathing cells from the ONL and peripheral olfactory fascicles. Furthermore, 1.9.E reacted with some processes of the radial glia in the periventricular germinal layer of the newborn rat. Although 4.11.C also specifically labeled ensheathing cells in the adult OB, it did not stain any cell type in the ONL of newborn rats. The lack of double labeling with either 1.9.E or 4.11.C and anti-olfactory marker protein (OMP) antibody, a specific marker for olfactory axons, indicated that none of the monoclonals recognized olfactory axons. Double immunostaining of adult OB cultures with 1.9.E or 4.11.C and anti-p75-nerve growth factor receptor revealed that both antibodies specifically recognized ensheathing glia in those cultures. Filaments were strongly labeled throughout the entire cytoplasm of ensheathing cells, suggesting that 1.9.E and 4.11.C immunoreacted with ensheathing glia cytoskeleton. 4.11.C stained a few Schwann cells in adult sciatic nerve sections. Moreover, 4.11.C immunostained cortical astrocyte cultures from newborn rats (P1). In Western blot analysis both antibodies recognized a major component, migrating with an apparent molecular weight of 60 kDa, from olfactory nerve and glomerular layer (ONGL) extracts of adult and neonatal rats. The pattern of immunoreactivity of 1.9.E and 4.11.C antibodies suggest that both antibodies are specific markers for olfactory ensheathing glia in the adult rat central nervous system (CNS).

Animals↗
PubMed1998

Olfactory ensheathing glia: properties and function.

The failure of regenerating axons to grow within the adult mammalian central nervous system (CNS) does not apply to the olfactory bulb (OB). In this structure, normal and transected olfactory axons are able to enter, regenerate, and reestablish lost synaptic contacts with their targets, throughout the lifetime of the organism. A remarkable difference between an axonal growth-permissive structure such as the OB and the remaining CNS resides in the presence of ensheathing glia in the former. These cells exhibit phenotypic and functional properties known to be involved in the process of axonal elongation that may explain the permissibility of the OB to axonal growth. In addition, transplants of ensheathing glia were successfully used to promote axonal regeneration within the injured adult CNS. The axonal growth-promoting properties of ensheathing glia make the study of this cell type interesting to provide an insight into the mechanisms underlying the process of axonal regeneration. Therefore, in this article we review the developmental, morphologic, immunocytochemical, and functional properties presented by this unique glial cell type, and correlate them with the axonal growth-promoting ability of ensheathing glia. In addition, we provide some evidence of the potentiality that ensheathing glia might have as a promoter of axonal regeneration within the injured nervous system.

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PubMed1997

Differential expression of microtubule-associated protein 1B phosphorylated isoforms in the adult rat nervous system.

Phosphorylated microtubule-associated protein 1B isoforms are thought to be involved in the plastic events taking place in neurons during development. However, little is known about their expression and possible role in the mature nervous system. To gain insight into the mechanisms underlying neuronal plasticity in the adult, we studied the pattern of expression of three microtubule-associated protein 1B isoforms in the entire adult rat nervous system. Accordingly, we performed western blots and immunohistochemistries using the antibodies 125, 150 and 531, which specifically recognize phosphorylated and unphosphorylated microtubule-associated protein 1B epitopes. Two electrophoretically distinct microtubule-associated protein 1B isoforms, slow-migrating and fast-migrating, were detected with the antibodies. The pattern of expression of these isoforms in the adult rat nervous system was region specific. Phosphorylated slow-migrating microtubule-associated protein 1B was expressed at all cellular compartments of primary sensory neurons in the central and peripheral nervous systems. In addition to primary sensory axons, slow-migrating microtubule-associated protein 1B was encountered at some other axons within the central nervous system. We discuss the correlation between slow-migrating microtubule-associated protein 1B axonal content and the regenerative potential of neurons. Phosphorylated fast-migrating microtubule-associated protein 1B was exclusively found in central nervous system dendrites where synaptic plasticity with morphological changes occurs in the adult. Unphosphorylated fast-migrating microtubule-associated protein 1B was the only isoform present in the bodies and dendrites of all motor neurons, and in peripheral and central nervous system glial cells of myelinated tracts with slow-migrating microtubule-associated protein 1B-containing axons. In summary, this report describes the pattern of expression of microtubule-associated protein 1B isoforms in the entire adult rat nervous system. In addition, it provides some information about the possible functional implications of phosphorylated microtubule-associated protein 1B isoforms in the adult.

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PubMed1995

Olfactory bulb ensheathing glia: a unique cell type with axonal growth-promoting properties.

The olfactory bulb (OB) is a structure of the central nervous system (CNS) in which axonal growth occurs throughout the lifetime of the organism. A major difference between the OB and the remaining CNS is the presence of ensheathing glia in the first two layers of the OB. Ensheathing glia display properties that might be involved in the process of regeneration and they appear to be responsible for the permissibility of the adult OB to axonal growth. In fact, transplants of ensheathing glia can be used as promoters of axonal regeneration within the adult CNS. The axonal growth-promoting properties of ensheathing glia make the study of this cell type interesting for understanding the mechanisms underlying axonal regeneration. Several groups have studied OB ensheathing cells extensively in an attempt to classify them within any of the known glial groups. However, this cell type does not exhibit the phenotypic features of any glial population described thus far. In this article we review the characteristics that differentiate ensheathing glia from other peripheral and central glial populations as well as the properties that involve them in axonal regeneration. The evidence suggests that ensheathing glia are unique, have their own identity, and do not belong to any previously described glial type.

Animals↗
PubMed1994

Regeneration into the spinal cord of transected dorsal root axons is promoted by ensheathing glia transplants.

The permissivity of adult olfactory bulb to the ingrowth of olfactory axons could be due to the unique properties of ensheathing glia. To test whether these glial cells could be used to promote axonal regeneration in a spontaneously nonregenerating system, we transplanted suspensions of pure ensheathing cells into a rhizotomized spinal cord segment. Ensheathing cells were purified away from other cell types by immunoaffinity, using anti-p75 nerve growth factor receptor. After laminectomy at the lower thoracic level, the spinal cord was exposed and one dorsal root (T10) was completely transected at the cord entry point. The root stump was microsurgically anastomosed to the cord and a suspension of ensheathing cells was transplanted in the spinal cord at the dorsal root entry zone. Three weeks after transplantation, numerous regenerating dorsal root axons were observed reentering the spinal cord. Ingrowth of dorsal root axons was observed using DiI and antibodies against calcitonin gene-related peptide and growth-associated protein. Primary sensory afferents invaded laminae 1, 2, and 3, grew through laminae 4 and 5, and reached the dorsal grey commissure and lamina 4 of the contralateral side. We did not observe regenerating axons within the ipsilateral ventral horn and dorsal column. Transplanted ensheathing cells reached the same laminae as axons. Neither ensheathing cells nor regenerating axons invaded those laminae they did not innervate under normal circumstances. In conclusion, the regeneration of injured dorsal root axons into the adult spinal cord was possible after ensheathing glia transplantation. The use of ensheathing cells as stimulators of axonal growth might be generalized to other central nervous system injuries.

Animals↗
PubMed1993

In vitro enfolding of olfactory neurites by p75 NGF receptor positive ensheathing cells from adult rat olfactory bulb.

Secondary cultures of adult rat olfactory bulb (OB) contained three different types of cell: (i) process-bearing cells; (ii) macrophage-like cells and (iii) fusiform cells. The immunohistochemical properties of process-bearing cells closely corresponded to those described for ensheathing glia in vivo. The most distinctive feature of these cells was their immunoreactivity for low affinity nerve growth factor receptor (NGFR). Process-bearing cells also shared the ultrastructural properties of ensheathing glia in vivo, as well as the ability to ensheath olfactory axons. In contrast, macrophage-like cells had the immunostaining properties of microglia, and fusiform cells were likely capillary endothelial cells. Neurites outgrowing from olfactory epithelium explants, when co-cultured with adult OB cells, grew preferentially over NGFR positive cells. Olfactory neurites exhibited NGFR immunoreactivity and were enfolded by NGFR positive cells. After ensheathment, this immunoreactivity decreased from the neurite and disappeared from the glial membrane in contact with the neurite. However, NGFR immunoreactivity was maintained in the portion of the glial membrane not involved in ensheathing. In summary, ensheathing cells in vitro retained both the ultrastructure shown in vivo and the ability to ensheath olfactory neurites. The Schwann cell-like properties of ensheathing glia, could partially explain the permissibility of adult OB to axonal growth.

Animals↗
PubMed1992

Glial cells from adult rat olfactory bulb: immunocytochemical properties of pure cultures of ensheathing cells.

Three morphologically and immunohistochemically distinct types of cell were present in primary cultures of adult rat olfactory nerve and glomerular layers of the olfactory bulb. One cell type was multipolar and stained positively for glial fibrillary acidic protein; a second type had fried egg-like morphology and stained with antibodies to epitope ED1; the third cell type had fusiform morphology, reacted with antibodies to vimentin and laminin and was glial fibrillary acidic protein- and ED1-negative. Trypsinization of these primary cultures (3 min, 37 degrees C), detached multipolar and fusiform cells only. When detached cells were set up in secondary culture on a glass substrate, fusiform cells did not attach, resulting in a pure culture of multipolar cells. Multipolar cells were glial fibrillary acidic protein- and myelin basic protein-positive and had the properties of so-called ensheathing cells or Blanes' glia. Immunoreactivity with anti-nerve growth factor receptor and anti-fibronectin allowed us to identify four distinct populations of multipolar ensheathing cells. One population was nerve growth factor receptor-positive, fibronectin-negative. A second was nerve growth factor receptor-negative and fibronectin-positive. A third was positive for both markers and the remaining cells did not stain for either of them. The morphological and immunological characteristics of cultured cells from olfactory nerve and glomerular layers were similar to those of Schwann cells and the similarities could account for the permissivity to axonal growth of the olfactory bulb.

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