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Biomedical subjects

M Nieto-Sampedro

Publications and source records attributed to M Nieto-Sampedro.

At least 19 recordsLinked to original sources
PubMed2004

Neural plasticity: changes with age.

Changes in the number, type and function of nervous system connections, in the morphology and function of glia and in neuron-glia interactions, are at the basis of vertebrate adjustment to changing environmental and physiological conditions. Collected under "neural plasticity", these age-dependent changes underlie adaptations apparently as different as the physiological response to dehydration or learning, and its electrophysiological and morphological correlates.

Aging↗
PubMed2004

Phrenic rehabilitation and diaphragm recovery after cervical injury and transplantation of olfactory ensheathing cells.

Functional respiratory recovery was evaluated by recording diaphragm and phrenic nerve activity several months after cervical cord hemisection followed by olfactory ensheathing cell (OEC) transplantation. The intact side was taken as a control in each rat. Sham-transplanted rats did not recover respiratory activity from the ipsilateral lesioned side. By contrast, ipsilateral phrenic and diaphragmatic activities recovered in transplanted rats amounted to 80.7% and 73% of their controls, respectively. After contralateral acute C1 section eliminating any contralateral influence from crossed compensatory pathways, the ipsilateral phrenic activity remained at 57.5% of the control, indicating that the phrenic recovery originated from the ipsilateral side. Supralesional stimulation in these rats elicited sublesional ipsilateral postsynaptic phrenic responses showing that transplantation helped ipsilateral fibers to again transmit nervous messages to the phrenic target, leading to substantial functional recovery. The origin of mechanisms involved in respiratory recovery (regeneration, resurrection, sprouting, sparing, demasking of latent pathways) is discussed.

Animals↗
PubMed2002

Spinal implants of olfactory ensheathing cells promote axon regeneration and bladder activity after bilateral lumbosacral dorsal rhizotomy in the adult rat.

PURPOSE: We performed spinal implantation of olfactory ensheathing cells to demonstrate dorsal root afferent regeneration as well as bladder activity restoration after lumbosacral L6 to S2 rhizotomy. MATERIALS AND METHODS: Spinal segments receiving bladder innervation, usually L6, S1 and S2, were identified by bipolar stimulation of the ventral roots. Bilateral section of the identified dorsal roots L6 to S2 was performed in 18 male Wistar rats. Immediately after rhizotomy olfactory ensheathing cells or vehicle was unilaterally injected in the vicinity of the sacral parasympathetic nucleus in 9 rats each using a glass micropipette and air pulse system. The severed roots were reattached to the cord with fibrin glue and the animals recovered under antibiotic prophylaxis. RESULTS: Anatomical regeneration of bladder wall primary afferents was demonstrated by the presence of labeled wheat germ agglutinin-horseradish peroxidase fibers in the dorsal horn and sacral parasympathetic nucleus in 8 of 9 cases of olfactory ensheathing cell implantation but not in the 9 controls injected with vehicle. One week after surgery all rats had an atonic bladder on cystometrography. At 6 weeks 8 of the 9 olfactory ensheathing cell implanted rats had recovered bladder activity. No recovery was observed in controls, in which vehicle was injected instead of olfactory ensheathing cells. CONCLUSIONS: Regenerated primary afferent fibers from the bladder project to the sacral parasympathetic nucleus, where they presumably form synapses mediating the recovery of bladder activity. Thus, olfactory ensheathing cell implants in the adult rat promote sensory axon regeneration, target reinnervation and bladder activity restoration.

Afferent Pathways↗
PubMed2001

Effects of ensheathing cells transplanted into photochemically damaged spinal cord.

Transplantation of olfactory ensheathing cells (OECs) into photochemically damaged rat spinal cord diminished astrocyte reactivity and parenchyma cavitation. The photochemical lesion performed at T12--L1 resulted in severe damage to the spinal cord, so that during the first 15 days postoperation all rats dragged their hindlimbs and did not respond to pinprick. The maximal area and volume of the cystic cavities were lower in transplanted than in non-transplanted rats, not significantly at the T12--L1 lesion site, but significantly at T9--T10 and L4--L6 cord levels. The density of astrocytes in the grey matter was similar at T12--L1 and L4--L6 in non-transplanted and trans- planted rats, but lower in the latter at T9--T10 level. However, in non-transplanted rats all astrocytes showed a hypertrophied appearance, with long and robust processes heavily GFAP-positive, and overexpression of proteoglycan inhibitor of neuritogenesis, whereas in transplanted rats only a few astrocytes showed hypertrophy and the majority had short, thin processes. These results indicate that OECs transplanted into damaged adult rat spinal cord exert a neuroprotective role by reducing astrocytic gliosis and cystic cavitation.

Animals↗
PubMed2001

Developmental and reactive growth of dentate gyrus afferents: cellular and molecular interactions.

The lamination of dentate gyrus afferents established during development is maintained following lesion-induced reactive growth in the adult. After partial deafferentation sprouts from undamaged afferents restore most synapses, while respecting the laminae relative boundaries. No evidence of trans-laminar sprouting has been found. Here, we review the information gathered during the last decade on the cellular and molecular bases of dentate synaptogenesis, with special attention to the role of glia during development and that of reactive glia after deafferentation. The interactions of neurons with astroglia and astroglial macromolecules, particularly proteoglycans, influence synapse segregation in the dentate gyrus, providing us with a reasonable explanation for afferent lamination.

Animals↗
PubMed2000

Expansion of adult Schwann cells from mouse predegenerated peripheral nerves.

We present an effective technique for culture and expansion of Schwann cells (SC) from adult peripheral nerves. Cultures from adult mouse sciatic nerves (one to six nerves per culture) in defined medium showed markedly higher purity and density of SC when the nerve was predegenerated in vivo for 7 days than when it was harvested fresh. SC from degenerated nerves were then cultured in defined media conditioned by primary cultures of adult SC. The best results were obtained with a conditioned medium supplemented with 1% fetal calf serum. In these conditions the purity of SC was about 90% and the density about 190 cell/mm(2) by 7-10 days in vitro. These findings indicate that adult SC can be expanded from small preinjured nerve fragments in a short time period to provide a source of SC for autologous cellular transplants.

Age Factors↗
PubMed2000

Limits to the capacity of transplants of olfactory glia to promote axonal regrowth in the CNS.

Olfactory bulb ensheathing cell (OBEC) transplants promoted axonal regeneration in the spinal cord dorsal root entry zone and in the corticospinal tract. However, OBECs failed to promote abducens internuclear neuron axon regeneration when transplanted at the site of nerve fibre transection. In experiments performed in both cats and rats, OBECs survived for up to 2 months, lining themselves up along the portion of the regrowing axons proximal to the interneuron cell body. However, OBECs migrated preferentially towards abducens somata, in the direction opposite to the oculomotor nucleus target. OBECs seem to promote nerve fibre regeneration only where preferred direction of glial migration coincides with the direction of axonal growth towards its target.

Abducens Nerve↗
PubMed2000

Schwann-like macroglia in adult rat brain.

Olfactory ensheathing cells (OECs) share properties with astrocytes and Schwann cells. This study was designed to test the hypothesis that glia with properties similar to those exhibited by OECs might be present in brain areas other than the olfactory bulb. We found tanycytes and pituicytes to express a distinctive set of immunological markers in common with OECs and nonmyelinating Schwann cells, namely low-affinity neurotrophin receptor (p75NTR), O4 antigen, estrogen receptor-alpha type, and insulin-like growth factor 1 (IGF-1). The two glial types could be cultured from adult hypothalamus and neurohypophysis, respectively, using the methods developed for olfactory OECs. Both glial types displayed morphologies reminiscent of Schwann cells, in primary culture. Schwann-like central glia presented a preferred growth substrate for dorsal root ganglion neurites and, when making intimate contacts with them, manifested a myelinating phenotype. These combined properties define a type of CNS macroglia that would not fit within conventional central glia types.

Animals↗
PubMed2000

Purification and structure of neurostatin, an inhibitor of astrocyte division of mammalian brain.

Neurostatin was originally described as an inhibitor of astroblast and astrocytoma division present in rat brain extracts and immunologically related to the sugar moiety of epidermal growth factor receptor and to blood group antigens. It was purified recently from mammalian brain extracts and characterized as a glycosphingolipid, but its precise structure remained unknown. Neurostatin has now been purified to apparent homogeneity from ganglioside extracts of rat, bovine, and porcine brain. It is cytostatic for astroblasts, C6 glioma cells, and various human astrocytomas grades III and IV, with IC(50) values ranging from 250 to 450 nM, but does not affect the division of primary or transformed fibroblasts up to concentrations >4 microM. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry of purified pig neurostatin showed a molecular ion of 1, 905 Da and ions of 1,863 and 1,934 Da, compatible with a disialoganglioside. Mono- and bidimensional NMR spectra, together with biochemical studies, suggest that neurostatin may be the 9-O-monoacetyl ester of GD1b.

3T3 Cells↗
PubMed1999

Estrogen receptor immunoreactivity in Schwann-like brain macroglia.

Olfactory ensheathing cells, tanycytes, pituicytes, pineal glia, retinal Müller cells, and Bergmann glia of normal male rats express concomitantly estrogen receptor, low-affinity neurotrophin receptor, antigen O4, and GFAP, markers characteristic of nonmyelinating Schwann cells. These cells were able to survive and proliferate when cultured from adult tissue, promoted neurite outgrowth, and could guide and ensheath growing neurites. We called this distinct group of growth-promoting central nervous system (CNS) macroglia aldynoglia (Greek: to make grow). Its proliferative and growth-promoting properties seem to be retained during the whole lifetime of the organism in those CNS loci where normal function depends on continuous axon renewal. Aldynoglia plasticity seems totally or partially lost with age where and when it is no longer critical, as in the case of adult cortical and spinal cord radial glia. The concomitant expression of estrogen receptor and low-affinity neurotrophin receptor may promote Schwann-like plasticity of glial cells.

Animals↗
PubMed1999

Olfactory bulb ensheathing cells enhance peripheral nerve regeneration.

Sciatic nerve resection leaving a 15 mm gap could not be repaired by bridging the stumps with a silicone tube prefilled with a laminin gel. However, when purified olfactory ensheathing cells (EC) were added to the gel filling the tube, successful axonal regeneration was observed in 50% of rats. With 12 mm gaps, regeneration occurred in 79% of rats with transplanted EC compared with 60% of those receiving collagen gel alone. Therefore, ECs help repair severe peripheral nerve injuries, in addition to their ability to promote axonal regeneration within the central nervous system.

Action Potentials↗
PubMed1999

Ensheathing glia transplants promote dorsal root regeneration and spinal reflex restitution after multiple lumbar rhizotomy.

Previously, we have shown that transplants of olfactory bulb ensheathing cells promoted regeneration of transected dorsal roots into the spinal cord. In this study, we assessed the ability of regenerating axons to make functional connections in the cord. Dorsal roots L3 to L6 were sectioned close to their entrance into the spinal cord and reapposed after injecting a suspension of ensheathing cells into each dorsal root entry zone (Group G). Afferent regeneration into the cord and recovery of spinal reflexes were compared with animals that received no injection (Group S) or culture medium without cells (Group C). Electrophysiological tests, to measure nerve conduction and spinal reflexes (H response and withdrawal reflex) evoked by stimulation of afferents of the sciatic nerve, were performed. At 14 days after surgery, H response was found in only 1 of 7 rats of Group G, and withdrawal reflexes were absent from all animals. At 60 days, the H response reappeared in 7 of 10 rats of Group G, and 1 of 5 of each of Groups C and S. The withdrawal reflex recovered in 4 of 10 rats of Group G, but in none of Groups C and S. Immunohistochemical labeling for calcitonin gene-related peptide (CGRP) in rats of Group G showed immunoreactive fibers entering the dorsal horn from sectioned roots, although at lower density than in the contralateral side. In conclusion, transplanted ensheathing cells promoted central regeneration and functional reconnection of regenerating sensory afferents.

Animals↗
PubMed1999

Neurite outgrowth inhibitors in gliotic tissue.

Gliotic tissue is the major obstacle to axon regeneration after CNS injury. We designed tissue culture assays to search for molecules responsible for neurite outgrowth inhibition in gliotic tissue. All the inhibitory activity in injured brain tissue was located in a plasma membrane heparan-sulphate and condroitin-sulphate type-proteoglycan of apparent molecular weight 200 kDalton. The proteoglycan core protein (apparent MW 48,000 kD) was biologically inactive, whereas the glycosamine-glycan (GAG) chains accounted for the inhibitory activity. Because of its cell location and mode of induction, the inhibitor was called injured membrane proteoglycan, IMP. IMP prevented neurite outgrowth initiation when attached to the culture substrate and caused growth cone collapse when added in solution to neurons with already growing neurites. We concluded that IMP was responsible for preventing injured CNS fibre regeneration. Double-staining immunohistochemistry of normal and gliotic tissue with anti-IMP monoclonal antibodies together with glial and neuronal markers, permitted the unequivocal definition of inhibitor presenting cells by confocal microscopy. IMP-immunostaining in normal CNS was observed exclusively on neurons. However, after a lesion, immunostaining occurred primarily on intensely GFAP-positive reactive astrocytes, but not on OX-42 positive microglia. The availability of antibodies permitted rapid affinity-purification of the neurite inhibitor and comparison with similar molecules possibly expressed during development. IMP itself or a highly related form, was expressed in embryonic brain, reaching maximal expression around postnatal day 3 and decreasing strongly in normal adult tissue. Perinatal rat brain proteoglycans inhibited neurite outgrowth similarly, though not identically, to IMP. Our data suggest that perinatal membrane and injured membrane proteoglycans may differ in GAG composition. IMP-like immunoreactivity was also found in developing brain, predominantly in neurons in normal brain, associating after a lesion with reactive astrocytes. Thes results suggest that injury evokes re-expression of IMP previously expressed during CNS development. One of the monoclonal antibodies to IMP blocked inhibitory activity, restoring neurite outgrowth in vitro. We are currently preparing Fab fragments to test the possibility that the antibody may block inhibition of central sprout growth in vivo. The combined use of blocking antibody fragments to neurite outgrowth inhibitors and transplants of growth-promoting glia, may help in the repair brain and spinal cord lesions.

Animals↗
PubMed1998

Novel disaccharide inhibitors of human glioma cell division.

Several alpha-L-Fuc-(1-->3)-alpha-D-GlcNAcOC8H17 disaccharide derivatives bearing different hydroxylated alkyl chains, with or without sulfate groups at C-4 and/or C-6 positions of the GlcNAc unit, have been synthesized and tested as inhibitors of human astrocytoma lines U-373 and U-118. The antimitotic activity was dependent on the structure and position of the hydroxylated chain linked to the disaccharide. The compounds with a pentaerythritol or L-glyceryl chain at the C-6 position showed the best inhibitory properties, with an ID50 value of ca. 200 microM. On the contrary, sulfated disaccharide derivatives were inactive. The antimitotic activities of the compounds tested were essentially independent of the mitogen used to stimulate cell division.

Antineoplastic Agents↗
PubMed1998

A neurite outgrowth-inhibitory proteoglycan expressed during development is similar to that isolated from adult brain after isomorphic injury.

The expression of proteoglycans (PGs) in the mammalian central nervous system (CNS) appears to be strictly regulated both during development and after damage to the mammalian CNS. Recently, we have isolated from membranes of injured adult brain a neurite outgrowth-inhibitory proteoglycan (IMP), the activity of which could be specifically counteracted by a monoclonal antibody (mAB) against the PG. We described in this report the characterization of perinatal membrane proteoglycan (PMP), a heparan-sulfate/chondroitin-sulfate-containing PG expressed during brain development. Its maximal expression was observed around postnatal day 3, decreasing strongly in normal adult tissue. This PG was purified and characterized using mABs generated against IMP. The comparison of PMP and IMP properties indicates that the two PGs are highly related and share expression patterns, biochemical characteristics, and the ability to inhibit neurite initiation in culture. However, IMP and PMP displayed a distinct effect on neurite elongation, which may be explained by their differences in glycosilation pattern. The data presented in this report support the idea that proteoglycans expressed during CNS development are re-expressed following injury.

Animals↗