RESUMEN
Anti-myelin basic protein (MBP) antibodies in pediatric-onset MS and controls were characterized. Serum samples were obtained from 94 children with MS and 106 controls. Paired CSF and serum were obtained from 25 children with MS at time of their initial episode of acute demyelinating syndrome (ADS). Complementary assays were applied across samples to evaluate the presence, and the physical binding properties, of anti-MBP antibodies. While the prevalence and titers of serum anti-MBP antibodies against both immature and mature forms of MBP were similar in children with MS and in controls, binding characteristics and formal Surface Plasmon Resonance (SPR) studies indicated surprisingly high binding affinities of all pediatric anti-MBP antibodies. Serum levels of anti-MBP antibodies correlated significantly with their CSF levels, and their presence in children with MS was associated with significantly increased risk of an acute disseminated encephalomyelitis-like initial clinical presentation. While antibodies to both immature and mature forms of MBP can be present as part of the normal pediatric humoral repertoire, these anti-myelin antibodies are of surprisingly high affinity, can access the CNS during inflammation, and have the capacity to modulate disease expression. Our findings identify an immune mechanism that could contribute to the observed heterogeneity in spectrum of clinical presentations in early-onset MS.
Asunto(s)
Autoanticuerpos/fisiología , Esclerosis Múltiple/diagnóstico , Esclerosis Múltiple/inmunología , Vaina de Mielina/inmunología , Proteínas del Tejido Nervioso/inmunología , Factores de Transcripción/inmunología , Enfermedad Aguda , Adolescente , Autoanticuerpos/sangre , Autoanticuerpos/líquido cefalorraquídeo , Biomarcadores/sangre , Biomarcadores/líquido cefalorraquídeo , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Esclerosis Múltiple/sangre , Esclerosis Múltiple/líquido cefalorraquídeo , Proteína Básica de Mielina , Proteínas del Tejido Nervioso/sangre , Proteínas del Tejido Nervioso/líquido cefalorraquídeo , Factores de Riesgo , Síndrome , Factores de Transcripción/sangre , Factores de Transcripción/líquido cefalorraquídeo , Adulto JovenRESUMEN
Nogo has been identified as a component of the central nervous system (CNS) myelin that prevents axonal regeneration in the adult vertebrate CNS. Analysis of Nogo-A has shown that an axon-inhibiting domain of 66 amino acids is expressed at the extracellular surface and at the endoplasmic reticulum lumen of transfected cells and oligodendrocytes. The acidic amino terminus of Nogo-A is detected at the cytosolic face of cellular membranes and may contribute to inhibition of axon regeneration at sites of oligodendrocyte injury. Here we show that the extracellular domain of Nogo (Nogo-66) inhibits axonal extension, but does not alter non-neuronal cell morphology. In contrast, a multivalent form of the N terminus of Nogo-A affects the morphology of both neurons and other cell types. Here we identify a brain-specific, leucine-rich-repeat protein with high affinity for soluble Nogo-66. Cleavage of the Nogo-66 receptor and other glycophosphatidylinositol-linked proteins from axonal surfaces renders neurons insensitive to Nogo-66. Nogo-66 receptor expression is sufficient to impart Nogo-66 axonal inhibition to unresponsive neurons. Disruption of the interaction between Nogo-66 and its receptor provides the potential for enhanced recovery after human CNS injury.
Asunto(s)
Axones/fisiología , Proteínas de la Mielina/fisiología , Regeneración Nerviosa , Receptores de Superficie Celular/metabolismo , Células 3T3 , Secuencia de Aminoácidos , Animales , Sitios de Unión , Células COS , División Celular , Línea Celular , Pollos , Clonación Molecular , ADN Complementario , Proteínas Ligadas a GPI , Expresión Génica , Conos de Crecimiento , Humanos , Ratones , Datos de Secuencia Molecular , Proteínas de la Mielina/química , Proteínas Nogo , Receptor Nogo 1 , Estructura Terciaria de Proteína , Receptores de Superficie Celular/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismoRESUMEN
During the past year, a major advance in the study of axon regeneration was the molecular cloning of Nogo. The expression of Nogo protein by CNS myelin may be a major factor in the failure of CNS axon regeneration. The effect of disrupting Nogo-dependent axon inhibition can now be studied conclusively. In related work, immunization with a Nogo-containing CNS myelin preparation was shown to promote regeneration and dramatic functional recovery after spinal cord trauma.
Asunto(s)
Axones/fisiología , Sistema Nervioso Central/fisiología , Factores de Crecimiento Nervioso/fisiología , Regeneración Nerviosa/fisiología , Sistema Nervioso Central/citología , Humanos , Proteínas de la Mielina/fisiología , Vaina de Mielina/fisiología , Proteínas NogoAsunto(s)
Conos de Crecimiento/fisiología , Proteínas de Unión al GTP rho/metabolismo , Animales , Células Cultivadas , Embrión de Pollo , Vectores Genéticos , Herpesvirus Humano 1/genética , Neuronas/citología , Neuronas Aferentes/citología , Células Ganglionares de la Retina/citología , Sistema Nervioso Simpático/citología , Transformación Genética , Proteínas de Unión al GTP rac/metabolismoRESUMEN
Axonal growth cone collapse is accompanied by a reduction in filopodial F-actin. We demonstrate here that semaphorin 3A (Sema3A) induces a coordinated rearrangement of Sema3A receptors and F-actin during growth cone collapse. Differential interference contrast microscopy reveals that some sites of Sema3A-induced F-actin reorganization correlate with discrete vacuoles, structures involved in endocytosis. Endocytosis of FITC-dextran by the growth cone is enhanced during Sema3A treatment, and sites of dextran accumulation colocalize with actin-rich vacuoles and ridges of membrane. Furthermore, the Sema3A receptor proteins, neuropilin-1 and plexin, and the Sema3A signaling molecule, rac1, also reorganize to vacuoles and membrane ridges after Sema3A treatment. These data support a model whereby Sema3A stimulates endocytosis by focal and coordinated rearrangement of receptor and cytoskeletal elements. Dextran accumulation is also increased in retinal ganglion cell (RGC) growth cones, in response to ephrin A5, and in RGC and DRG growth cones, in response to myelin and phorbol-ester. Therefore, enhanced endocytosis may be a general principle of physiologic growth cone collapse. We suggest that growth cone collapse is mediated by both actin filament rearrangements and alterations in membrane dynamics.
Asunto(s)
Actinas/fisiología , Glicoproteínas/farmacología , Factores de Crecimiento Nervioso/farmacología , Proteínas del Tejido Nervioso/fisiología , Neuronas/citología , Neuronas/fisiología , Actinas/efectos de los fármacos , Animales , Moléculas de Adhesión Celular/fisiología , Membrana Celular/efectos de los fármacos , Membrana Celular/fisiología , Membrana Celular/ultraestructura , Embrión de Pollo , Endocitosis/efectos de los fármacos , Endocitosis/fisiología , Ganglios Espinales/citología , Microscopía de Interferencia/métodos , Proteínas del Tejido Nervioso/efectos de los fármacos , Neuronas/ultraestructura , Neuropilina-1 , Receptores de Superficie Celular/fisiología , Retina/embriología , Semaforina-3A , Transducción de Señal , Proteína de Unión al GTP rac1/fisiologíaRESUMEN
Axonal regrowth after injury is accompanied by changes in the expression of tubulin, but the contributions of substrate molecules and neurotrophic factors in regulating these changes in vivo are not known. Adult rat retinal ganglion cells (RGCs) were examined after intraorbital axotomy, after application of a peripheral nerve (PN) graft to stimulate regeneration, and after axotomy and treatment with brain-derived neurotrophic factor (BDNF). After these treatments we used in situ hybridization to study mRNA levels for betaI, betaII, betaIII, betaIVa, and Talpha1 tubulin isotypes. Levels of mRNA for all isotypes were downregulated after intraorbital axotomy. During regrowth of injured RGC axons, mRNA levels for betaII, betaIII, and Talpha1 isotypes were upregulated specifically and dramatically, suggesting that elevated expression of these isotypes is correlated specifically with axonal regrowth. A corresponding increase in betaIII protein levels was detected by immunocytochemistry. The betaI and betaIVa mRNAs were not increased during regeneration. BDNF did not elicit a specific increase in the mRNA levels for the betaIII and Talpha1 isotypes and had only a small effect on mRNA levels for the betaII isotype. Therefore, despite the ability of BDNF to support the survival of injured RGCs and to enhance neurite outgrowth of retinal neurons in vitro, the in vivo application of BDNF alone is unable to induce the program of changes in growth-associated tubulins that accompany regeneration of RGC axons into PN grafts. We speculate that, in addition to BDNF, cooperative signaling with substrate molecules is required to allow RGCs to regenerate and exhibit tubulin isotype switching.
Asunto(s)
Axones/fisiología , Factor Neurotrófico Derivado del Encéfalo/farmacología , Regeneración Nerviosa , Nervio Óptico/fisiología , Células Ganglionares de la Retina/fisiología , Nervio Ciático/fisiología , Nervio Ciático/trasplante , Transcripción Genética , Tubulina (Proteína)/biosíntesis , Animales , Femenino , Hibridación in Situ , Traumatismos del Nervio Óptico , ARN Mensajero/biosíntesis , Ratas , Ratas Sprague-Dawley , Células Ganglionares de la Retina/efectos de los fármacos , Trasplante AutólogoRESUMEN
The administration of neurotrophins affects neuronal survival and growth, but less is known about their ability to modify the expression of growth associated genes following injury to CNS neurons. Here we characterize the effect of brain-derived neurotrophic factor (BDNF) on mRNA levels for T alpha1 alpha-tubulin, and for GAP-43, two genes whose expression levels in retinal ganglion cells (RGC) tend to correlate with growth. We first determined that most adult rat RGCs can retrogradely transport BDNF by injecting 125I-BDNF into RGC target sites in vivo. We then used quantitative in situ hybridization to characterize the effect of axotomy, or axotomy and BDNF administration on mRNA levels for GAP-43 and T alpha1. Axotomy alone resulted in a general decrease in T alpha1 alpha-tubulin mRNA levels by 2 weeks, and elicited an increase in GAP-43 mRNA levels in an average of 30% of surviving RGCs. The intravitreal administration of a single dose of BDNF (5 microg) to axotomized RGCs on the day of injury did not affect T alpha1 alpha-tubulin mRNA levels, but was followed by a moderate (approximately 80%), and short-lasting enhancement of GAP-43 mRNA levels in most RGCs during the first week after axotomy. No significant increase in GAP-43 mRNA levels was observed when BDNF was injected into the uninjured eye. We conclude that BDNF specifically enhances GAP-43 but not T alpha1 mRNA levels in injured RGCs. Because BDNF is known to stimulate branch length of injured RGCs, we suggest that changes in the expression of GAP-43, but not T alpha1 tubulin, correlate with branching of injured neurons as opposed to long distance regrowth.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/farmacología , Glicoproteínas de Membrana/biosíntesis , Proteínas del Tejido Nervioso/biosíntesis , Nervio Óptico/fisiología , Células Ganglionares de la Retina/fisiología , Transcripción Genética/efectos de los fármacos , Tubulina (Proteína)/biosíntesis , Animales , Autorradiografía , Transporte Axonal , Factor Neurotrófico Derivado del Encéfalo/farmacocinética , Femenino , Proteína GAP-43 , Hibridación in Situ , Radioisótopos de Yodo , Proteínas de Neurofilamentos/biosíntesis , Traumatismos del Nervio Óptico , ARN Mensajero/biosíntesis , Ratas , Ratas Sprague-Dawley , Proteínas Recombinantes/farmacocinética , Proteínas Recombinantes/farmacología , Valores de Referencia , Células Ganglionares de la Retina/efectos de los fármacosRESUMEN
Microtubules are essential components of the cytoskeleton required for axonal growth. To investigate how changes in tubulin transport and expression may affect axon regeneration, injury in the adult mammalian central nervous system was studied. Axotomized retinal ganglion cells (RGCs) that do not regenerate were compared with RGCs that regenerate their axons when the optic nerve is replaced with a peripheral nerve graft. When RGC axons regenerated through peripheral nerve grafts, the rate of slow transport increased but decreased when no regrowth occurred. To investigate the molecular mechanisms that mediate these responses, alterations in tubulin mRNA levels after injury were examined. Total tubulin mRNA levels fell after injury in the optic nerve but increased in those RGCs that regenerated their axons into a peripheral nerve graft. Further, the expression of four separate beta-tubulin isotypes in injured rat RGCs was characterized. mRNA levels for all four isotypes decreased in RGCs after injury in the optic nerve. How the autoregulation of tubulin expression may contribute to the changes in beta-tubulin isotype expression after injury is discussed.
Asunto(s)
Transporte Axonal/fisiología , Sistema Nervioso Central/fisiología , Mamíferos/fisiología , Regeneración Nerviosa/fisiología , Células Ganglionares de la Retina/fisiología , Tubulina (Proteína)/biosíntesis , Animales , Sistema Nervioso Central/citología , Sistema Nervioso Central/lesiones , Mamíferos/metabolismo , Células Ganglionares de la Retina/metabolismoAsunto(s)
Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/tratamiento farmacológico , Animales , Huesos/metabolismo , Calcio/metabolismo , Calcio/uso terapéutico , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/metabolismo , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/terapia , Humanos , Absorción Intestinal , Glándulas Paratiroides/cirugía , Hormona Paratiroidea/metabolismo , Fósforo/metabolismo , Ratas , Diálisis Renal , Vitamina D/análogos & derivados , Vitamina D/metabolismo , Vitamina D/uso terapéuticoAsunto(s)
Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/tratamiento farmacológico , Hidroxicolecalciferoles/uso terapéutico , Adulto , Fosfatasa Alcalina/sangre , Resorción Ósea , Calcificación Fisiológica/efectos de los fármacos , Calcio/metabolismo , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/metabolismo , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/patología , Ensayos Clínicos como Asunto , Femenino , Humanos , Absorción Intestinal/efectos de los fármacos , Masculino , Persona de Mediana Edad , Hormona Paratiroidea/sangre , Fosfatos/sangreRESUMEN
Four microgrammes of 1-alpha-hydroxycholecalciferol (1-alpha-OH D3) or 200 mug of 25-hydroxycholecalciferol (25-OH D3) were given orally every other day respectively to 10 uraemic patients (8 on chronic haemodialysis) for 1-12 weeks and to 3 patients on chronic haemodialysis for 4-8 weeks. A transilial bone biopsy and serial evaluation of serum immunoreactive PTH (iPTH) calcium phosphate and alkaline phosphatase were performed before and at the end of therapy. Both 1-alpha-OH D3 and 25-OH D3 (the latter at a 50 times higher dose) were able to depress hyperparathyroidism in two-thirds of the cases and to consistently improve the mineralisation defect. In no case did iPTH or the bone histomorphometric parameters return to normal, so that long term evaluation of these two drugs is warranted.
Asunto(s)
Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/tratamiento farmacológico , Hidroxicolecalciferoles/uso terapéutico , Absorción , Adulto , Fosfatasa Alcalina/sangre , Antígenos , Desarrollo Óseo/efectos de los fármacos , Resorción Ósea/tratamiento farmacológico , Calcificación Fisiológica/efectos de los fármacos , Calcio/metabolismo , Fosfatos de Calcio/sangre , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/sangre , Femenino , Humanos , Hidroxicolecalciferoles/farmacología , Masculino , Persona de Mediana Edad , Hormona Paratiroidea/inmunología , Fosfatos/sangre , Uremia/sangre , Uremia/tratamiento farmacológicoRESUMEN
Parathyroid function was assessed by calcium infusions (4-8 h) in 16 patients with chronic renal insufficiency being treated by long-term hemodialysis. The concentrations of two immunoreactive species of parathyroid hormone in plasma (iPTH-9, mol wt 9500; iPTH-7, mol wt 7000) were estimated by radioimmunoassays utilizing two relatively specific antisera. Control values of the smaller species, iPTH-7, were uniformly high, whereas values of iPTH-9 were normal in 12 of 19 studies. Response of iPTH-7 to calcium infusions was variable, with significant decreases occurring only five times in 27 infusions. Concentrations of iPTH-9, however, decreased during every calcium infusion. In contrast to these acute responses, five of six patients studied during periods of dialysis against both low (< 6 mg/100 ml) and high (7-8 mg/100 ml) calcium concentrations in the dialyzate showed a decrease in values of iPTH-7 during the period of dialysis against the higher calcium concentration. It is concluded that plasma concentrations of iPTH-9 reflect primarily the moment-to-moment secretory status of the parathyroid glands, while concentrations of iPTH-7 reflect more closely chronic parathyroid functional status. It is further concluded that the failure of iPTH-7 to decrease during induced hypercalcemia should not be equated with autonomy of parathyroid gland function.
Asunto(s)
Enfermedades Óseas/etiología , Hiperparatiroidismo/etiología , Diálisis Renal/efectos adversos , Animales , Calcio/administración & dosificación , Calcio/sangre , Bovinos/inmunología , Cromatografía en Gel , Gluconatos/administración & dosificación , Humanos , Sueros Inmunes , Isótopos de Yodo , Fallo Renal Crónico/terapia , Métodos , Peso Molecular , Glándulas Paratiroides/fisiopatología , Hormona Paratiroidea/sangre , Fósforo/sangre , Radioinmunoensayo , Espectrofotometría Atómica , Porcinos/inmunología , Factores de TiempoAsunto(s)
Hiperparatiroidismo/terapia , Diálisis Renal/efectos adversos , Administración Oral , Aluminio/administración & dosificación , Calcio/administración & dosificación , Calcio/sangre , Femenino , Humanos , Hidróxidos/administración & dosificación , Hiperparatiroidismo/complicaciones , Hiperparatiroidismo/etiología , Fallo Renal Crónico/terapia , Masculino , Osteítis Fibrosa Quística/etiología , Fosfatos/sangreRESUMEN
The present study was prompted by the observation that, in patients with chronic renal failure being followed at this center, renal osteodystrophy developed almost exclusively in those who were treated by chronic hemodialysis at home rather than in our center. A systematic comparison was made between the 10 patients with roentgenographic evidence of the bone disease and 18 patients without demonstrable bone disease. The two groups were similar in age, sex, nature of renal disease, and duration of dialysis. The mean duration of kidney disease was almost 2 yr longer in the patients without bone disease than in those with bone disease. Other significant differences related to where the hemodialysis was performed and to the calcium concentration in the dialysate (6.0-7.4 mg/100 ml in the hospital and 4.9-5.6 mg/100 ml at home). If the unknown factors related to where the dialysis was performed were of no consequence, the major factor contributing to the production of bone disease observed in these patients was the use of a dialysate with a calcium concentration less than 5.7 mg/100 ml.
Asunto(s)
Enfermedades Óseas/etiología , Adulto , Fosfatasa Alcalina/sangre , Proteínas Sanguíneas/análisis , Calcinosis/sangre , Calcinosis/etiología , Calcio/administración & dosificación , Calcio/sangre , Calcio de la Dieta , Creatina/sangre , HumanosRESUMEN
Plasma concentration of immunoreactive parathyroid hormone (IPTH) was measured in 18 patients who had been on a hemodialysis program for longer than 6 months. A negative correlation was found between the predialysis plasma concentration of IPTH and the mean concentration of calcium in the dialysate previously used: plasma concentrations of IPTH were higher in patients dialyzed against a calcium concentration between 4.9 and 5.6 mg/100 ml than in patients dialyzed against a calcium concentration of 6.0 mg/100 ml or more. Plasma concentrations of IPTH also were higher in patients with bone disease than in patients without bone disease. Furthermore, a positive correlation was found between predialysis plasma concentrations of IPTH and calcium, and between mean predialysis concentration of IPTH and phosphate. To obviate the possibility that individual differences in susceptibility could have accounted for the observed effects of plasma phosphate and of dialysate calcium, a 2 x 2 factorial study was conducted in seven of these patients to examine the independent effects of perturbation of each of these factors. It was observed that plasma concentration of IPTH was lowest with the combination of high dialysate calcium and low plasma phosphate, highest with the combination of low dialysate calcium and high plasma phosphate, and intermediate with the two other combinations. It is concluded that both dialysate calcium and plasma phosphate are important determinants of parathyroid function in these patients.