RESUMEN
Current therapies for multiple sclerosis (MS) are largely palliative, not curative. Mesenchymal stem cells (MSCs) harbor regenerative and immunosuppressive functions, indicating a potential therapy for MS, yet the variability and low potency of MSCs from adult sources hinder their therapeutic potential. MSCs derived from human embryonic stem cells (hES-MSCs) may be better suited for clinical treatment of MS because of their unlimited and stable supply. Here, we show that hES-MSCs significantly reduce clinical symptoms and prevent neuronal demyelination in a mouse experimental autoimmune encephalitis (EAE) model of MS, and that the EAE disease-modifying effect of hES-MSCs is significantly greater than that of human bone-marrow-derived MSCs (BM-MSCs). Our evidence also suggests that increased IL-6 expression by BM-MSCs contributes to the reduced anti-EAE therapeutic activity of these cells. A distinct ability to extravasate and migrate into inflamed CNS tissues may also be associated with the robust therapeutic effects of hES-MSCs on EAE.
Asunto(s)
Células de la Médula Ósea/citología , Células Madre Embrionarias/citología , Encefalomielitis Autoinmune Experimental/terapia , Células Madre Mesenquimatosas/citología , Esclerosis Múltiple/patología , Esclerosis Múltiple/terapia , Animales , Sistema Nervioso Central/patología , Modelos Animales de Enfermedad , Humanos , Trasplante de Células Madre Mesenquimatosas , RatonesRESUMEN
Globoid cell leukodystrophy (GLD) or Krabbe disease, is a fatal demyelinating disease attributed to mutations in the galactocerebrosidase (GALC) gene. Loss of function mutations in GALC result in accumulation of the glycolipid intermediate, galactosylsphingosine (psychosine). Due to the cytotoxicity of psychosine, it has been hypothesized that accumulated psychosine underlie the pathophysiology of GLD. However, the cellular mechanisms of GLD pathophysiology remain unclear. Globoid cells, multinucleated microglia/macrophages in the central nervous system (CNS), are a defining characteristic of GLD. Here we report that exposure of primary glial cultures to psychosine induces the expression and the production of matrix metalloproteinase (MMP)-3 that mediated a morphological transformation of microglia into a multinucleated globoid cell type. Additionally, psychosine-induced globoid cell formation from microglia was prevented by either genetic ablation or chemical inhibition of MMP-3. These effects are microglia-specific as peripheral macrophages exposed to psychosine did not become activated or express increased levels of MMP-3. In the brain from twitcher mice, a murine model of human GLD, elevated MMP-3 expression relative to wild-type littermates was contemporaneous with disease onset and further increased with disease progression. Further, bone marrow transplantation (BMT), currently the only therapeutically beneficial treatment for GLD, did not mitigate the elevated expression of MMP-3 in twitcher mice. Hence, elevated expression of MMP-3 in GLD may promote microglial responses to psychosine that may represent an important pathophysiological process in this disease and its treatment.
Asunto(s)
Leucodistrofia de Células Globoides/enzimología , Leucodistrofia de Células Globoides/patología , Metaloproteinasa 3 de la Matriz/fisiología , Psicosina/toxicidad , Animales , Animales Recién Nacidos , Células Cultivadas , Leucodistrofia de Células Globoides/inducido químicamente , Ratones , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
The pannexin-1 (Panx1) channel (often referred to as the Panx1 hemichannel) is a large-conductance channel in the plasma membrane of many mammalian cells. While opening of the channel is potentially detrimental to the cell, little is known about how it is regulated under physiological conditions. Here we show that stomatin inhibited Panx1 channel activity. In transfected HEK-293 cells, stomatin reduced Panx1-mediated whole-cell currents without altering either the total or membrane surface Panx1 protein expression. Stomatin coimmunoprecipitated with full-length Panx1 as well as a Panx1 fragment containing the fourth membrane-spanning domain and the cytosolic carboxyl terminal. The inhibitory effect of stomatin on Panx1-mediated whole-cell currents was abolished by truncating Panx1 at a site in the cytosolic carboxyl terminal. In primary culture of mouse astrocytes, inhibition of endogenous stomatin expression by small interfering RNA enhanced Panx1-mediated outward whole-cell currents. These observations suggest that stomatin may play important roles in astrocytes and other cells by interacting with Panx1 carboxyl terminal to limit channel opening.
Asunto(s)
Proteínas Sanguíneas/metabolismo , Conexinas/química , Conexinas/metabolismo , Activación del Canal Iónico , Proteínas de la Membrana/metabolismo , Proteínas del Tejido Nervioso/química , Proteínas del Tejido Nervioso/metabolismo , Animales , Astrocitos/metabolismo , Proteínas Sanguíneas/antagonistas & inhibidores , Membrana Celular/metabolismo , Etidio/metabolismo , Células HEK293 , Humanos , Inmunoprecipitación , Proteínas de la Membrana/antagonistas & inhibidores , Ratones , Ratones Endogámicos C57BL , Unión Proteica , Relación Estructura-Actividad , TransfecciónRESUMEN
Transcription factor COUP-TFII in rodents is important for migration of cortical interneurons from caudal ganglionic eminence (CGE) to the neocortex. Since in human, unlike in rodents, cortical interneurons have both ganglionic eminence (GE) and dorsal cortical origin, we studied the distribution of COUP-TFII in the human developing neocortex from 9 to 22 gestational weeks. COUP-TFII is expressed at all stages studied in the GE and in various cortical zones, from the proliferative ventricular/subventricular zone (VZ/SVZ) to layer I. Gradients of COUP-TFII expression are present in the GE, with peak expression in the CGE, and in the neocortex, from high expression in the temporal and occipital cortex to moderate in the frontal and dorsal cortex. Double immunofluorescence with γ-aminobutyric acid (GABA), calretinin, or calbindin, established that subpopulations of interneurons express COUP-TFII. A small fraction of COUP-TFII(+) cells are progenitor cells that proliferate in the CGE (3.4 ± 0.3%) and in the cortical VZ/SVZ (1.7 ± 0.1%). In summary, COUP-TFII is expressed in the human fetal forebrain in GABAergic cells, according to its possible role in migration of cortical interneurons. The source of these cells seems to be the CGE and, to a smaller extent, the cortical VZ/SVZ.
Asunto(s)
Factor de Transcripción COUP II/metabolismo , Regulación del Desarrollo de la Expresión Génica/fisiología , Interneuronas/metabolismo , Prosencéfalo/embriología , Prosencéfalo/metabolismo , Femenino , Humanos , Técnicas In Vitro , Masculino , Prosencéfalo/citología , Distribución TisularRESUMEN
The viewpoint aftereffect is a perceptual illusion that, after adapting to an object/face viewed from one side (e.g., 30 degrees to the left of center), when the same object/face is subsequently presented near the front view, the perceived viewing direction is biased in a direction opposite to that of the adapted viewpoint (e.g., 2 degrees to the right). In this study, we measured the face viewpoint aftereffects when the adapting and the testing faces were different in identity and gender and when their vertical orientations were inverted. The aftereffect showed a strong transfer following adaptation to other faces. This effect was slightly attenuated when the adapting and the test face stimuli were made more dissimilar. This suggests the existence of neurons jointly tuned to both face view and structure. However, the transfer from cross adapting to an inverted face was much reduced and weak, indicating that the neural coding of upright and inverted faces in the high-level visual cortex is different and a major part of the face viewpoint coding occurs at the level where faces are holistically represented.