RESUMEN
Mice with insulin receptor (IR)-deficient astrocytes (GFAP-IR knockout [KO] mice) show blunted responses to insulin and reduced brain glucose uptake, whereas IR-deficient astrocytes show disturbed mitochondrial responses to glucose. While exploring the functional impact of disturbed mitochondrial function in astrocytes, we observed that GFAP-IR KO mice show uncoupling of brain blood flow with glucose uptake. Since IR-deficient astrocytes show higher levels of reactive oxidant species (ROS), this leads to stimulation of hypoxia-inducible factor-1α and, consequently, of the vascular endothelial growth factor angiogenic pathway. Indeed, GFAP-IR KO mice show disturbed brain vascularity and blood flow that is normalized by treatment with the antioxidant N-acetylcysteine (NAC). NAC ameliorated high ROS levels, normalized angiogenic signaling and mitochondrial function in IR-deficient astrocytes, and normalized neurovascular coupling in GFAP-IR KO mice. Our results indicate that by modulating glucose uptake and angiogenesis, insulin receptors in astrocytes participate in neurovascular coupling.
Asunto(s)
Astrocitos , Encéfalo , Insulina , Neovascularización Fisiológica , Acoplamiento Neurovascular , Animales , Astrocitos/metabolismo , Encéfalo/irrigación sanguínea , Proteína Ácida Fibrilar de la Glía/genética , Glucosa/metabolismo , Insulina/metabolismo , Ratones , Ratones Noqueados , Especies Reactivas de Oxígeno/metabolismo , Receptor de Insulina/genética , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
BACKGROUND: The complex pathophysiology of Alzheimer's disease (AD) hampers the development of effective treatments. Attempts to prevent neurodegeneration in AD have failed so far, highlighting the need for further clarification of the underlying cellular and molecular mechanisms. Neuroinflammation seems to play a crucial role in disease progression, although its specific contribution to AD pathogenesis remains elusive. We have previously shown that the modulation of the endocannabinoid system (ECS) renders beneficial effects in a context of amyloidosis, which triggers neuroinflammation. In the 5xFAD model, the genetic inactivation of the enzyme that degrades anandamide (AEA), the fatty acid amide hydrolase (FAAH), was associated with a significant amelioration of the memory deficit. METHODS: In this work, we use electrophysiology, flow cytometry and molecular analysis to evaluate the cellular and molecular mechanisms underlying the improvement associated to the increased endocannabinoid tone in the 5xFAD mouse- model. RESULTS: We demonstrate that the chronic enhancement of the endocannabinoid tone rescues hippocampal synaptic plasticity in the 5xFAD mouse model. At the CA3-CA1 synapse, both basal synaptic transmission and long-term potentiation (LTP) of synaptic transmission are normalized upon FAAH genetic inactivation, in a CB1 receptor (CB1R)- and TRPV1 receptor-independent manner. Dendritic spine density in CA1 pyramidal neurons, which is notably decreased in 6-month-old 5xFAD animals, is also restored. Importantly, we reveal that the expression of microglial factors linked to phagocytic activity, such as TREM2 and CTSD, and other factors related to amyloid beta clearance and involved in neuron-glia crosstalk, such as complement component C3 and complement receptor C3AR, are specifically upregulated in 5xFAD/FAAH-/- animals. CONCLUSION: In summary, our findings support the therapeutic potential of modulating, rather than suppressing, neuroinflammation in Alzheimer's disease. In our model, the long-term enhancement of the endocannabinoid tone triggered augmented microglial activation and amyloid beta phagocytosis, and a consequent reversal in the neuronal phenotype associated to the disease.
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Enfermedad de Alzheimer/metabolismo , Amidohidrolasas/deficiencia , Péptidos beta-Amiloides/metabolismo , Plasticidad Neuronal/fisiología , Transmisión Sináptica/fisiología , Enfermedad de Alzheimer/patología , Animales , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fagocitosis/fisiologíaRESUMEN
NMDA receptor-dependent long-term depression (LTD) in the hippocampus is a well-known form of synaptic plasticity that has been linked to different cognitive functions. The core mechanism for this form of plasticity is thought to be entirely neuronal. However, we now demonstrate that astrocytic activity drives LTD at CA3-CA1 synapses. We have found that LTD induction enhances astrocyte-to-neuron communication mediated by glutamate, and that Ca2+ signaling and SNARE-dependent vesicular release from the astrocyte are required for LTD expression. In addition, using optogenetic techniques, we show that low-frequency astrocytic activation, in the absence of presynaptic activity, is sufficient to induce postsynaptic AMPA receptor removal and LTD expression. Using cell-type-specific gene deletion, we show that astrocytic p38α MAPK is required for the increased astrocytic glutamate release and astrocyte-to-neuron communication during low-frequency stimulation. Accordingly, removal of astrocytic (but not neuronal) p38α abolishes LTD expression. Finally, this mechanism modulates long-term memory in vivo.
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Astrocitos/enzimología , Hipocampo/fisiología , Memoria a Largo Plazo/fisiología , Proteína Quinasa 14 Activada por Mitógenos/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Animales , Conducta Animal/fisiología , Condicionamiento Psicológico/fisiología , Miedo/fisiología , Femenino , Ácido Glutámico/metabolismo , Hipocampo/citología , Depresión Sináptica a Largo Plazo/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Neuronas/fisiología , Optogenética , Técnicas de Placa-Clamp , Potenciales Sinápticos/fisiologíaRESUMEN
A correction to this article has been published and is linked from the HTML version of this paper. The error has been fixed in the paper.
RESUMEN
Pyramidal neuron subtypes differ in intrinsic electrophysiology properties and dendritic morphology. However, do different pyramidal neuron subtypes also receive synaptic inputs that are dissimilar in frequency and in excitation/inhibition balance? Unsupervised clustering of three intrinsic parameters that vary by cell subtype - the slow afterhyperpolarization, the sag, and the spike frequency adaptation - split layer 5 barrel cortex pyramidal neurons into two clusters: one of adapting cells and one of non-adapting cells, corresponding to previously described thin- and thick-tufted pyramidal neurons, respectively. Non-adapting neurons presented frequencies of spontaneous inhibitory postsynaptic currents (sIPSCs) and spontaneous excitatory postsynaptic currents (sEPSCs) three- and two-fold higher, respectively, than those of adapting neurons. The IPSC difference between pyramidal subtypes was activity independent. A subset of neurons were thy1-GFP positive, presented characteristics of non-adapting pyramidal neurons, and also had higher IPSC and EPSC frequencies than adapting neurons. The sEPSC/sIPSC frequency ratio was higher in adapting than in non-adapting cells, suggesting a higher excitatory drive in adapting neurons. Therefore, our study on spontaneous synaptic inputs suggests a different extent of synaptic information processing in adapting and non-adapting barrel cortex neurons, and that eventual deficits in inhibition may have differential effects on the excitation/inhibition balance in adapting and non-adapting neurons.
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Corteza Cerebral/fisiología , Potenciales Postsinápticos Inhibidores , Células Piramidales/fisiología , Transmisión Sináptica , Animales , Corteza Cerebral/citología , Corteza Cerebral/ultraestructura , Potenciales Postsinápticos Excitadores , Femenino , Masculino , Ratones Transgénicos , Células Piramidales/citología , Células Piramidales/ultraestructuraRESUMEN
The regulated transport of AMPA-type glutamate receptors (AMPARs) to the synaptic membrane is a key mechanism to determine the strength of excitatory synaptic transmission in the brain. In this work, we uncovered a new role for the microtubule-associated protein MAP1B in modulating access of AMPARs to the postsynaptic membrane. Using mice and rats of either sex, we show that MAP1B light chain (LC) accumulates in the somatodendritic compartment of hippocampal neurons, where it forms immobile complexes on microtubules that limit vesicular transport. These complexes restrict AMPAR dendritic mobility, leading to the intracellular trapping of receptors and impairing their access to the dendritic surface and spines. Accordingly, increasing MAP1B-LC expression depresses AMPAR-mediated synaptic transmission. This effect is specific for the GluA2 subunit of the AMPAR and requires glutamate receptor interacting protein 1 (GRIP1) interaction with MAP1B-LC. Therefore, MAP1B-LC represents an alternative link between GRIP1-AMPARs and microtubules that does not result in productive transport, but rather limits AMPAR availability for synaptic insertion, with a direct impact on synaptic transmission.SIGNIFICANCE STATEMENT The ability of neurons to modify their synaptic connections, known as synaptic plasticity, is accepted as the cellular basis for learning and memory. One mechanism for synaptic plasticity is the regulated addition and removal of AMPA-type glutamate receptors (AMPARs) at excitatory synapses. In this study, we found that a microtubule-associated protein, MAP1B light chain (MAP1B-LC), participates in this process. MAP1B-LC forms immobile complexes along dendrites. These complexes limit intracellular vesicular trafficking and trap AMPARs inside the dendritic shaft. In this manner, MAP1B restricts the access of AMPARs to dendritic spines and the postsynaptic membrane, contributing to downregulating synaptic transmission.
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Proteínas Asociadas a Microtúbulos/fisiología , Receptores AMPA/fisiología , Sinapsis/fisiología , Transmisión Sináptica/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Animales , Dendritas/efectos de los fármacos , Espinas Dendríticas/fisiología , Femenino , Hipocampo/citología , Hipocampo/metabolismo , Masculino , Ratones , Proteínas Asociadas a Microtúbulos/biosíntesis , Proteínas Asociadas a Microtúbulos/genética , Proteínas del Tejido Nervioso/metabolismo , Neuronas/metabolismo , Ratas , Ratas Wistar , Receptores AMPA/metabolismoRESUMEN
Alzheimer's disease (AD) is a neurodegenerative disorder characterized by a severe and progressive neuronal loss leading to cognitive dysfunctions. Previous reports, based on the use of chemical inhibitors, have connected the stress kinase p38α to neuroinflammation, neuronal death and synaptic dysfunction. To explore the specific role of neuronal p38α signalling in the appearance of pathological symptoms, we have generated mice that combine expression of the 5XFAD transgenes to induce AD symptoms with the downregulation of p38α only in neurons (5XFAD/p38α∆-N). We found that the neuronal-specific deletion of p38α improves the memory loss and long-term potentiation impairment induced by 5XFAD transgenes. Furthermore, 5XFAD/p38α∆-N mice display reduced amyloid-ß accumulation, improved neurogenesis, and important changes in brain cytokine expression compared with 5XFAD mice. Our results implicate neuronal p38α signalling in the synaptic plasticity dysfunction and memory impairment observed in 5XFAD mice, by regulating both amyloid-ß deposition in the brain and the relay of this accumulation to mount an inflammatory response, which leads to the cognitive deficits.
Asunto(s)
Enfermedad de Alzheimer/psicología , Péptidos beta-Amiloides/metabolismo , Disfunción Cognitiva/metabolismo , Proteína Quinasa 14 Activada por Mitógenos/genética , Neuronas/metabolismo , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/metabolismo , Animales , Células Cultivadas , Citocinas/metabolismo , Modelos Animales de Enfermedad , Regulación hacia Abajo , Eliminación de Gen , Humanos , Ratones , Ratones Transgénicos , Proteína Quinasa 14 Activada por Mitógenos/deficiencia , Plasticidad Neuronal , Neuronas/citología , Transducción de SeñalRESUMEN
El uso de catéteres centrales ha aumentando de forma importante a pesar de su mayor morbi-mortalidad en comparación con las FAV. Las principales complicaciones son trombosis, bacteriemia-sepsis y limitación de la eficacia dialítica disminuyendo la calidad de vida de los pacientes y elevando el coste sanitario. En nuestra unidad, el sellado se realizaba con heparina sódica al 5% y actualmente utilizamos TauroLockTM Hep 500(R), compuesta de (ciclo)-taurolidina (agente antimicrobiano biocompatible), citrato al 4% y heparina 500 UI/ml. Objetivo: comprobar la eficacia del Taurolock para reducir la incidencia de colonización intraluminal y trombosis al compararla con heparina sódica al 5%. Material y método: realizamos un estudio observacional longitudinal de cohorte y analizamos colonización intraluminal, trombosis, biofilm y uso de antibioterapia. Resultados: observamos mejoría en la permeabilidad del catéter, disminución del uso de urokinasa, reducción de sesiones en unipunción y del uso de antibiótico. Conclusiones: nuestro estudio muestra que el sellado del catéter para hemodiálisis con una solución que contiene taurolidina como antimicrobiano puede reducir significativamente la incidencia de sepsis relacionada con el catéter. La taurolidina parece se efectiva y segura y no conlleva riesgos ni efectos secundarios. La adicción de 500 ui de heparina a la solución de sellado junto con citrato al 4% ha disminuido los eventos trombóticos reduciendo el uso de fibrilolíticos y el uso de unipunción totalmente, presentando una mejoría de la eficacia dialítica
The use of central catheters has increased significantly despite their greater morbi-mortality compared to AVF. The main complications are thrombosis, bacteriaemiasepsis, and limitation of dialysis efficacy, reducing quality of life for patients and increasing the healthcare cost. In our unit, 5% heparin sodium was previously used as lock solution and we currently use TauroLockTM Hep 500(R), composed of (cyclo)-taurolidine (biocompatible antimicrobial agent) 4% citrate and heparin 500 UI/ml. Objective: to verify the efficacy of Taurolock in reducing the incidence of intraluminal colonization and thrombosis compared with 5% heparin sodium. Material and method: we carried out a longitudinal observational cohort study and we analysed intraluminal colonization, thrombosis, biofilm and use of antibiotherapy. Results: we observed an improvement in catheter permeability, a reduction in the use of urokinase, reduction in unipuncture sessions and in use of antibiotic. Conclusions: our study shows that using a lock solution that contains taurolidine as an antimicrobial agent for haemodialysis catheters can significantly reduce the incidence of catheter-related sepsis. Taurolidine appears to be effective and safe and does not entail risks or side effects. The addition of 500 iu of heparin to the lock solution together with 4% citrate has reduced thrombotic events by reducing the use of fibrilolytics and eliminating completely the use of unipuncture, presenting an improvement in dialysis efficacy
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Humanos , Cateterismo/normas , Obstrucción del Catéter/estadística & datos numéricos , Catéteres Venosos Centrales/estadística & datos numéricos , Anticoagulantes/uso terapéutico , Hemostáticos/análisis , Infecciones Relacionadas con Catéteres/prevención & control , Catéteres de Permanencia/efectos adversos , Heparina/uso terapéutico , Diálisis Renal/métodos , Insuficiencia Renal Crónica/terapiaRESUMEN
The microtubule-associated protein 1B (MAP1B) plays critical roles in neurite growth and synapse maturation during brain development. This protein is well expressed in the adult brain. However, its function in mature neurons remains unknown. We have used a genetically modified mouse model and shRNA techniques to assess the role of MAP1B at established synapses, bypassing MAP1B functions during neuronal development. Under these conditions, we found that MAP1B deficiency alters synaptic plasticity by specifically impairing long-term depression (LTD) expression. Interestingly, this is due to a failure to trigger AMPA receptor endocytosis and spine shrinkage during LTD. These defects are accompanied by an impaired targeting of the Rac1 activator Tiam1 at synaptic compartments. Accordingly, LTD and AMPA receptor endocytosis are restored in MAP1B-deficient neurons by providing additional Rac1. Therefore, these results indicate that the MAP1B-Tiam1-Rac1 relay is essential for spine structural plasticity and removal of AMPA receptors from synapses during LTD. This work highlights the importance of MAPs as signalling hubs controlling the actin cytoskeleton and receptor trafficking during plasticity in mature neurons.