RESUMEN
The occurrence of major depressive disorder (MDD) has been linked to an increased vulnerability to stress. The basolateral amygdala (BLA) is one of the critical brain areas that involved in the regulation of pathological reactivity to stress. Increasing evidence indicates that the EphB2 receptor (EphB2) plays a critical role in neuropsychiatric disorders, such as Alzheimer's disease, pain and anxiety. However, whether the EphB2 in the BLA is involved in stress vulnerability is unclear. Here, we identified EphB2 in the BLA as a key regulator contributed to the modulation of stress vulnerability in adult mice. We found that the expression of EphB2 in the BLA was significantly increased in the animal model induced by chronic social stress. Knockdown of EphB2 in the BLA produced antidepressant-like behavioral effects, whereas activation of EphB2 in the BLA increased the susceptibility to subthreshold social defeat stress. Furthermore, we demonstrated that the role of EphB2 in the stress vulnerability was mediated by modulating NMDA receptors, since the knockdown of EphB2 in the BLA prevented not only the increase in the amplitudes of both the miniature and the evoked NMDAR-mediated EPSC, but also the enhancement of surface expression of NMDARs in the defeated mice. Taken together, these results suggest that EphB2 in the BLA is a critical factor contributes to the vulnerability to stress, which may be a potential target for the treatment of depression.
Asunto(s)
Complejo Nuclear Basolateral/metabolismo , Receptor EphB2/biosíntesis , Receptores de N-Metil-D-Aspartato/biosíntesis , Interacción Social , Estrés Psicológico/metabolismo , Sinapsis/metabolismo , Animales , Complejo Nuclear Basolateral/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Receptor EphB2/deficiencia , Interacción Social/efectos de los fármacos , Estrés Psicológico/psicología , Sinapsis/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiología , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiónico/farmacologíaAsunto(s)
Puente de Arteria Coronaria , Transportadores de Anión Orgánico/genética , Receptor EphB2/metabolismo , Vena Safena/cirugía , Miembro 1B3 de la Familia de los Transportadores de Solutos de Aniones Orgánicos/genética , Animales , Células Epiteliales/citología , Células Epiteliales/metabolismo , Humanos , Islotes Pancreáticos/metabolismo , Ratones , Transportadores de Anión Orgánico/análisis , Receptor EphB2/deficiencia , Vena Safena/patología , Miembro 1B3 de la Familia de los Transportadores de Solutos de Aniones Orgánicos/análisis , Timocitos/citología , Timocitos/patología , Timo/citología , Timo/metabolismoRESUMEN
Epithelial-free areas, present in both thymic cortex and medulla, have been studied in WT and EphB-deficient mice that have important alterations in the development of thymic epithelium due to the lack of proper thymocyte-thymic epithelial cell interactions. In both WT and mutant thymuses, the number and size of epithelial-free areas are significantly larger in the medulla than in the cortex. The two parameters show a reverse correlation: low numbers of these areas course with large epithelial-free areas and vice versa. However, their structure and cell content are similar in mutant and WT thymuses. Cortical epithelial-free areas just contain DP thymocytes, while the medullary ones consist of SP cells, blood vessels, mesenchyme-derived ER-TR7+ cells and components of the extracellular matrix (i.e., collagen IV, fibronectin, laminin). Other components, such as desmin, αSMA, PDGFRß and Ng2, frequently associated with blood vessel walls, also appear. Vimentin, although present in medullary epithelial-free areas, does not co-express with epithelial cells. Other markers related to epithelial-mesenchymal transitions, such as Snail, Slug or FSP1, are not expressed. These results suggest that alterations in the cell interactions between distinct thymic cell components that induce both increased proportions of apoptotic thymic epithelial cells and altered behavior of the mesenchyme associated with the medullary vasculature could explain the appearance of these areas and their differences in the cortex and medulla.
Asunto(s)
Células Epiteliales/metabolismo , Receptor EphB2/metabolismo , Receptor EphB3/metabolismo , Timocitos/metabolismo , Timo/metabolismo , Animales , Células Epiteliales/citología , Transición Epitelial-Mesenquimal , Femenino , Masculino , Ratones , Ratones Noqueados , Receptor EphB2/deficiencia , Receptor EphB3/deficiencia , Timocitos/citología , Timo/citologíaRESUMEN
The development of complex organs such as the eye requires a delicate and coordinated balance of cell division and cell death. Although apoptosis is prevalent in the proximoventral optic cup, the precise role it plays in eye development needs to be investigated further. In this study, we show that reduced apoptosis in the proximoventral optic cup prevents closure of the optic fissure. We also show that expression of ephrin A5 (Efna5) partially overlaps with Eph receptor B2 (Ephb2) expression in the proximoventral optic cup and that binding of EphB2 to ephrin A5 induces a sustained activation of JNK. This prolonged JNK signal promotes apoptosis and prevents cell proliferation. Thus, we propose that the unique cross-subclass interaction of EphB2 with ephrin A5 has evolved to function upstream of JNK signaling for the purpose of maintaining an adequate pool of progenitor cells to ensure proper closure of the optic fissure.
Asunto(s)
Efrina-A5/metabolismo , Sistema de Señalización de MAP Quinasas , Disco Óptico/embriología , Disco Óptico/metabolismo , Receptor EphB2/metabolismo , Animales , Apoptosis/efectos de los fármacos , Membrana Celular/metabolismo , Proliferación Celular/efectos de los fármacos , Embrión de Mamíferos/metabolismo , Activación Enzimática , Efrina-A5/deficiencia , Células HEK293 , Humanos , Ratones Transgénicos , Modelos Biológicos , Morfogénesis , Receptor EphB2/deficiencia , Transducción de SeñalRESUMEN
The Eph receptor tyrosine kinases interact with their ephrin ligands on adjacent cells to facilitate contact-dependent cell communication. Ephrin B ligands are expressed on T cells and have been suggested to act as co-stimulatory molecules during T cell activation. There are no detailed reports of the expression and modulation of EphB receptors on dendritic cells, the main antigen presenting cells that interact with T cells. Here we show that mouse splenic dendritic cells (DC) and bone-marrow derived DCs (BMDC) express EphB2, a member of the EphB family. EphB2 expression is modulated by ligation of TLR4 and TLR9 and also by interaction with ephrin B ligands. Co-localization of EphB2 with MHC-II is also consistent with a potential role in T cell activation. However, BMDCs derived from EphB2 deficient mice were able to present antigen in the context of MHC-II and produce T cell activating cytokines to the same extent as intact DCs. Collectively our data suggest that EphB2 may contribute to DC responses, but that EphB2 is not required for T cell activation. This result may have arisen because DCs express other members of the EphB receptor family, EphB3, EphB4 and EphB6, all of which can interact with ephrin B ligands, or because EphB2 may be playing a role in another aspect of DC biology such as migration.
Asunto(s)
Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Regulación de la Expresión Génica , Activación de Linfocitos/inmunología , Receptor EphB2/genética , Linfocitos T/inmunología , Receptores Toll-Like/metabolismo , Animales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Línea Celular , Citocinas/biosíntesis , Femenino , Antígenos de Histocompatibilidad Clase II/inmunología , Antígenos de Histocompatibilidad Clase II/metabolismo , Humanos , Mediadores de Inflamación/metabolismo , Ligandos , Activación de Linfocitos/genética , Ratones , Ratones Noqueados , Unión Proteica , Receptor EphB2/deficiencia , Receptor EphB2/metabolismo , Linfocitos T/metabolismoRESUMEN
OBJECTIVES/HYPOTHESIS: Determine if differences in right and left semicircular size account for phenotypic behavior, indicating vestibulopathy in EphB deficient mice. STUDY DESIGN: The diameters of the superior semicircular canals (SCC) were measured. The differences in the right and left superior SCC diameters were analyzed in homozygous EphB2 and EphB3 double knockout mice known to have head bobbing and circling behavior. Results were compared to similar analysis in wild type controls that displayed no signs of vestibulopathy. METHODS: Axial frozen sections through the superior (SCC) were analyzed by light microscopy; and the diameters of the left and right canals were measured in µm for both EphB2 and EphB3 double knockout mice, as well as in wild type control mice. The differences in diameter between the left and right superior SCC was determined for each animal. RESULTS: Overall, the EphB2 and EphB3 double knockout mice had smaller superior SCC diameters compared to wild type (109.0±21.4 µm vs. 185.0±5.2 µm (P<0.0001). The mean difference in left and right diameter of the superior SCC of EphB2/EphB3 double knockout mice was 29.0±8.7 µm; in wild-type controls this difference was 6.0±5.1 µm (P=0.002). In addition, the direction of circling appeared to be independent of the laterality of the smaller (or larger) superior SCC. CONCLUSION: Mice deficient in EphB2/EphB3 signaling have smaller superior SCC and asymmetry in lumen sizes between the left and right sides. The laterality of the larger versus smaller is not correlated with the direction of circling behavior. LEVEL OF EVIDENCE: N/A.
Asunto(s)
Conducta Animal , Receptor EphB2/deficiencia , Canales Semicirculares/patología , Enfermedades Vestibulares/psicología , Animales , Modelos Animales de Enfermedad , Ratones , Ratones Noqueados , Fenotipo , Receptor EphB2/genética , Canales Semicirculares/fisiopatología , Transducción de Señal , Enfermedades Vestibulares/metabolismo , Enfermedades Vestibulares/patologíaRESUMEN
Precision in auditory brainstem connectivity underlies sound localization. Cochlear activity is transmitted to the ventral cochlear nucleus (VCN) in the mammalian brainstem via the auditory nerve. VCN globular bushy cells project to the contralateral medial nucleus of the trapezoid body (MNTB), where specialized axons terminals, the calyces of Held, encapsulate MNTB principal neurons. The VCN-MNTB pathway is an essential component of the circuitry used to compute interaural intensity differences that are used for localizing sounds. When input from one ear is removed during early postnatal development, auditory brainstem circuitry displays robust anatomical plasticity. The molecular mechanisms that control the development of auditory brainstem circuitry and the developmental plasticity of these pathways are poorly understood. In this study we examined the role of EphB signaling in the development of the VCN-MNTB projection and in the reorganization of this pathway after unilateral deafferentation. We found that EphB2 and EphB3 reverse signaling are critical for the normal development of the projection from VCN to MNTB, but that successful circuit assembly most likely relies upon the coordinated function of many EphB proteins. We have also found that ephrin-B reverse signaling repels induced projections to the ipsilateral MNTB after unilateral deafferentation, suggesting that similar mechanisms regulate these two processes.
Asunto(s)
Vías Auditivas/embriología , Vías Auditivas/crecimiento & desarrollo , Tronco Encefálico/embriología , Tronco Encefálico/crecimiento & desarrollo , Receptor EphB2/fisiología , Receptor EphB3/fisiología , Vías Aferentes/fisiología , Animales , Vías Auditivas/citología , Tronco Encefálico/citología , Núcleo Coclear/citología , Núcleo Coclear/embriología , Núcleo Coclear/crecimiento & desarrollo , Femenino , Masculino , Ratones , Ratones de la Cepa 129 , Ratones Noqueados , Ratones Transgénicos , Neurogénesis/fisiología , Receptor EphB2/deficiencia , Receptor EphB2/genética , Receptor EphB3/deficiencia , Receptor EphB3/genética , Transducción de Señal/fisiologíaAsunto(s)
Enfermedad de Alzheimer/fisiopatología , Enfermedad de Alzheimer/terapia , Hipocampo/fisiología , Hipocampo/fisiopatología , Memoria/fisiología , Receptor EphB2/metabolismo , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Animales , Giro Dentado/metabolismo , Modelos Animales de Enfermedad , Humanos , Potenciación a Largo Plazo/fisiología , Ratones , Plasticidad Neuronal , Receptor EphB2/biosíntesis , Receptor EphB2/deficiencia , Receptores de N-Metil-D-Aspartato/metabolismo , Sinapsis/metabolismoRESUMEN
Amyloid-ß oligomers may cause cognitive deficits in Alzheimer's disease by impairing neuronal NMDA-type glutamate receptors, whose function is regulated by the receptor tyrosine kinase EphB2. Here we show that amyloid-ß oligomers bind to the fibronectin repeats domain of EphB2 and trigger EphB2 degradation in the proteasome. To determine the pathogenic importance of EphB2 depletions in Alzheimer's disease and related models, we used lentiviral constructs to reduce or increase neuronal expression of EphB2 in memory centres of the mouse brain. In nontransgenic mice, knockdown of EphB2 mediated by short hairpin RNA reduced NMDA receptor currents and impaired long-term potentiation in the dentate gyrus, which are important for memory formation. Increasing EphB2 expression in the dentate gyrus of human amyloid precursor protein transgenic mice reversed deficits in NMDA receptor-dependent long-term potentiation and memory impairments. Thus, depletion of EphB2 is critical in amyloid-ß-induced neuronal dysfunction. Increasing EphB2 levels or function could be beneficial in Alzheimer's disease.
Asunto(s)
Enfermedad de Alzheimer/fisiopatología , Enfermedad de Alzheimer/terapia , Cognición/fisiología , Receptor EphB2/deficiencia , Receptor EphB2/metabolismo , Péptidos beta-Amiloides/metabolismo , Animales , Línea Celular , Células Cultivadas , Giro Dentado/metabolismo , Modelos Animales de Enfermedad , Humanos , Potenciación a Largo Plazo , Memoria/fisiología , Ratones , Ratones Transgénicos , Plasticidad Neuronal , Complejo de la Endopetidasa Proteasomal/metabolismo , Unión Proteica , Estructura Terciaria de Proteína , Ratas , Receptor EphB2/química , Receptor EphB2/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Sinapsis/metabolismoRESUMEN
In the current study, we extend our own previous results on the thymocyte phenotype of EphB2 and/or EphB3 deficient mice by analyzing the phenotype and the histological organization of their thymic epithelial stroma. All studied adult EphB-deficient thymi showed profound alterations with respect to the wild-type (WT) ones. Each mutant exhibited a specific phenotype, but also showed common features including occurrence of K5+K8+MTS10+ immature medullary epithelial cells, numerous K5-K8-MTS20+ cells and K5+K8+ cells in the thymic cortex and cortical and medullary K5-K8- areas devoid of epithelial cell markers. In addition, comparative analysis of WT and EphB-deficient embryonic and newborn thymi demonstrated that the observed adult phenotype was a consequence of the gradual accumulation of early phenotypic and morphological defects, becoming more severe at the end of embryonic life and in newborn animals. Together, these results confirm a role for EphB2 and EphB3 in thymus morphogenesis. The obtained data are discussed from the point of view of the recognized role played by these two Ephs in the homeostasis of other epithelia and their possible relationships with molecules known to be involved in thymic epithelial cell development.
Asunto(s)
Receptor EphB2/fisiología , Receptor EphB3/fisiología , Timo/embriología , Timo/crecimiento & desarrollo , Animales , Animales Recién Nacidos , Epitelio/anomalías , Epitelio/embriología , Epitelio/crecimiento & desarrollo , Epitelio/fisiología , Femenino , Antígenos de Histocompatibilidad Clase II/metabolismo , Queratina-15 , Queratina-5/metabolismo , Queratina-8/metabolismo , Laminina/metabolismo , Ratones , Ratones Noqueados , Morfogénesis/genética , Morfogénesis/fisiología , Fenotipo , Embarazo , Receptor EphB2/deficiencia , Receptor EphB2/genética , Receptor EphB3/deficiencia , Receptor EphB3/genética , Transducción de Señal , Timo/anomalías , Timo/fisiologíaRESUMEN
Ephs and ephrins are cell surface receptors that bind to each other and initiate distinct, bidirectional signalling pathways in processes known as forward (Eph) and reverse (ephrin) signalling. Previous work had shown that the loss of ephrinB1 protein alone or compound loss of EphB2 and EphB3 leads to cleft palate. Because of the bidirectional signalling capability of these molecules, it was not clear whether forward or reverse signalling caused the cleft palate in the ephrinB1 protein null or EphB2 and EphB3 compound null mice. We demonstrate that forward signalling is essential for palatogenesis. Foetuses with a cytoplasmically truncated EphB2 protein, which could initiate reverse but not forward signalling, and were protein null for EphB3 had a cleft palate. This happened because their palatal shelves, which could elevate in vivo and adhere and fuse in culture, were too small to contact one another. Small shelf size was due to reduced proliferation in the palatal mesenchyme. The reduced proliferation was not the result of abnormal vascular development within the palate. In conclusion, strong evidence is provided for specific and co-operative roles of EphB2 and EphB3 in palate development.
Asunto(s)
Efrina-B3/metabolismo , Hueso Paladar/embriología , Hueso Paladar/metabolismo , Receptor EphB2/metabolismo , Transducción de Señal , Adhesividad , Animales , Apoptosis , Recuento de Células , Proliferación Celular , Fisura del Paladar/embriología , Fisura del Paladar/patología , Efrina-B3/deficiencia , Matriz Extracelular/metabolismo , Femenino , Feto/anomalías , Feto/metabolismo , Masculino , Mesodermo/citología , Mesodermo/metabolismo , Ratones , Hueso Paladar/patología , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Receptor EphB2/deficiencia , beta-Galactosidasa/metabolismoRESUMEN
EphB receptor tyrosine kinases and ephrin-B ligands regulate several types of cell-cell interactions during brain development, generally by modulating the cytoskeleton. EphB/ephrinB genes are expressed in the developing neural tube of early mouse embryos with distinct overlapping expression in the ventral midbrain. To test EphB function in midbrain development, mouse embryos compound homozygous for mutations in the EphB2 and EphB3 receptor genes were examined for early brain phenotypes. These mutants displayed a morphological defect in the ventral midbrain, specifically an expanded ventral midline evident by embryonic day E9.5-10.5, which formed an abnormal protrusion into the cephalic flexure. The affected area was comprised of cells that normally express EphB2 and ephrin-B3. A truncated EphB2 receptor caused a more severe phenotype than a null mutation, implying a dominant negative effect through interference with EphB forward (intracellular) signaling. In mutant embryos, the overall number, size, and identity of the ventral midbrain cells were unaltered. Therefore, the defect in ventral midline morphology in the EphB2;EphB3 compound mutant embryos appears to be caused by cellular changes that thin the tissue, forcing a protrusion of the ventral midline into the cephalic space. Our data suggests a role for EphB signaling in morphological organization of specific regions of the developing neural tube.
Asunto(s)
Mesencéfalo/embriología , Mesencéfalo/metabolismo , Receptor EphB2/metabolismo , Receptor EphB3/metabolismo , Animales , Tipificación del Cuerpo , Proliferación Celular , Regulación del Desarrollo de la Expresión Génica/genética , Ratones , Mutación/genética , Receptor EphB2/deficiencia , Receptor EphB2/genética , Receptor EphB3/deficiencia , Receptor EphB3/genéticaRESUMEN
Here, using a genetic approach, we dissect the roles of EphB receptor tyrosine kinases in dendritic spine development. Analysis of EphB1, EphB2, and EphB3 double and triple mutant mice lacking these receptors in different combinations indicates that all three, although to varying degrees, are involved in dendritic spine morphogenesis and synapse formation in the hippocampus. Hippocampal neurons lacking EphB expression fail to form dendritic spines in vitro and they develop abnormal spines in vivo. Defective spine formation in the mutants is associated with a drastic reduction in excitatory glutamatergic synapses and the clustering of NMDA and AMPA receptors. We show further that a kinase-defective, truncating mutation in EphB2 also results in abnormal spine development and that ephrin-B2-mediated activation of the EphB receptors accelerates dendritic spine development. These results indicate EphB receptor cell autonomous forward signaling is responsible for dendritic spine formation and synaptic maturation in hippocampal neurons.