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Voltage-gated CaV2.1 (P/Q-type) Ca2+ channels play a crucial role in regulating neurotransmitter release, thus contributing to synaptic plasticity and to processes such as learning and memory. Despite their recognized importance in neural function, there is limited information on their potential involvement in neurodegenerative conditions such as Alzheimer's disease (AD). Here, we aimed to explore the impact of AD pathology on the density and nanoscale compartmentalization of CaV2.1 channels in the hippocampus in association with GABAB receptors. Histoblotting experiments showed that the density of CaV2.1 channel was significantly reduced in the hippocampus of APP/PS1 mice in a laminar-dependent manner. CaV2.1 channel was enriched in the active zone of the axon terminals and was present at a very low density over the surface of dendritic tree of the CA1 pyramidal cells, as shown by quantitative SDS-digested freeze-fracture replica labelling (SDS-FRL). In APP/PS1 mice, the density of CaV2.1 channel in the active zone was significantly reduced in the strata radiatum and lacunosum-moleculare, while it remained unaltered in the stratum oriens. The decline in Cav2.1 channel density was found to be associated with a corresponding impairment in the GABAergic synaptic function, as evidenced by electrophysiological experiments carried out in the hippocampus of APP/PS1 mice. Remarkably, double SDS-FRL showed a co-clustering of CaV2.1 channel and GABAB1 receptor in nanodomains (~40-50 nm) in wild type mice, while in APP/PS1 mice this nanoarchitecture was absent. Together, these findings suggest that the AD pathology-induced reduction in CaV2.1 channel density and CaV2.1-GABAB1 de-clustering may play a role in the synaptic transmission alterations shown in the AD hippocampus. Therefore, uncovering these layer-dependent changes in P/Q calcium currents associated with AD pathology can benefit the development of future strategies for AD management.
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The cannabinoid system is one of the most investigated neuromodulatory systems because of its involvement in multiple pathologies such as cancer, inflammation, and psychiatric diseases. Recently, the CB2 receptor has gained increased attention considering its crucial role in modulating neuroinflammation in several pathological conditions like neurodegenerative diseases. Here we describe the rational design of pyrrole-based analogues, which led to a potent and pharmacokinetically suitable CB2 full agonist particularly effective in improving cognitive functions in a scopolamine-induced amnesia murine model. Therefore, we extended our study by investigating the interconnection between CB2 activation and neurotransmission in this experimental paradigm. To this purpose, we performed a MALDI imaging analysis on mice brains, observing that the administration of our lead compound was able to revert the effect of scopolamine on different neurotransmitter tones, such as acetylcholine, serotonin, and GABA, shedding light on important networks not fully explored, so far.
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Cannabinoides , Receptor Cannabinoide CB2 , Ratones , Animales , Pirroles/farmacología , Cannabinoides/farmacología , Neurotransmisores/farmacología , Derivados de Escopolamina , Agonistas de Receptores de Cannabinoides/farmacología , Receptor Cannabinoide CB1RESUMEN
OBJECTIVES: Cannabidiol (CBD) is a phytocannabinoid with great potential in clinical applications. The mechanism(s) of action of CBD require further investigation. Previous studies suggested that adenosine A2A receptors (A2ARs) could play a role in CBD-induced effects. Here, we evaluated the ability of CBD to modify the function of A2AR. METHODS: We used HEK-293T cells transfected with the cDNA encoding the human A2AR and Gαs protein, both modified to perform bioluminescence-based assays. We first assessed the effect of CBD on A2AR ligand binding using an A2AR NanoLuciferase sensor. Next, we evaluated whether CBD modified A2AR coupling to mini-Gαs proteins using the NanoBiT™ assay. Finally, we further assessed CBD effects on A2AR intrinsic activity by recording agonist-induced cAMP accumulation. RESULTS: CBD did not bind orthosterically to A2AR but reduced the coupling of A2AR to Gαs protein and the subsequent generation of cAMP. CONCLUSION: CBD negatively modulates A2AR functioning.
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In recent years, there have been important legislative changes in many countries regarding the use of cannabis for medicinal and/or recreational purposes, which have facilitated access to it. Uruguay, Canada and some of the US states are the only jurisdictions that have legalised recreational consumption, applying different legislative models. The aim of this review is to analyse the effects that the legalisation of recreational cannabis has had on its use and its consequences. In general, the evidence accumulated to date indicates that the legalisation of cannabis has been associated with a decrease in the price of the substance, higher concentration of THC (potency), greater diversity of presentations for consumption, lower risk perception and an increase in consumption in adults and moderately in adolescents (even though it is illegal for them to consume), as well as an increase in the adverse consequences derived from cannabis consumption on public health. There has been a decrease in drug-related arrests, but the illegal market continues to be frequently used. No increase in the demand for treatment due to cannabis consumption has been detected. Therefore, these legislative changes have so far failed to achieve their main objectives, which were to suppress the illegal market and protect the most vulnerable groups, while on the contrary, they seem to imply an increase in some of the negative aspects associated with cannabis consumption. However, taking into account that most of these legislative changes have entered into force relatively recently, a longer follow-up period is required to be able to draw definitive conclusions.
En los últimos años se han producido importantes cambios legislativos en numerosos países respecto al consumo de cannabis con fines medicinales y/o recreativos, que han facilitado su accesibilidad. Actualmente, Uruguay, Canadá y algunos estados de EE.UU. han legalizado el consumo recreativo, aplicando distintos modelos legislativos. El objetivo de la presente revisión es analizar los efectos que ha tenido la legalización del cannabis recreativo sobre su consumo y sus consecuencias. En general, las evidencias indican que la legalización se ha asociado a un descenso en el precio, mayor concentración de THC (potencia), mayor diversidad de presentaciones para su consumo, una menor percepción de riesgo y un incremento en el consumo en adultos y de forma moderada en adolescentes (aunque sea ilegal el consumo para ellos), así como un aumento de las consecuencias adversas derivadas del consumo en la salud pública. Se ha producido un descenso en los arrestos relacionados con el consumo, pero el mercado ilegal sigue utilizándose de forma habitual. No se ha detectado un incremento de la demanda de tratamiento por este consumo. Por el momento, estos cambios legislativos no han conseguido alcanzar sus objetivos principales que eran suprimir el mercado ilegal y proteger a los grupos más vulnerables, mientras que, por el contrario, parecen implicar un incremento de algunos aspectos negativos asociados al consumo de cannabis. Sin embargo, teniendo en cuenta que la mayoría de estos cambios legislativos han entrado en vigor hace relativamente poco tiempo, se requiere un periodo de seguimiento mayor para poder extraer conclusiones definitivas.
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Cannabis , Adulto , Adolescente , Humanos , Cannabis/efectos adversos , UruguayRESUMEN
BACKGROUND AND PURPOSE: Opioid-based drugs are the gold standard medicines for pain relief. However, tolerance and several side effects (i.e. constipation and dependence) may occur upon chronic opioid administration. Photopharmacology is a promising approach to improve the benefit/risk profiles of these drugs. Thus, opioids can be locally activated with high spatiotemporal resolution, potentially minimizing systemic-mediated adverse effects. Here, we aimed at developing a morphine photo-derivative (photocaged morphine), which can be activated upon light irradiation both in vitro and in vivo. EXPERIMENTAL APPROACH: Light-dependent activity of pc-morphine was assessed in cell-based assays (intracellular calcium accumulation and electrophysiology) and in mice (formalin animal model of pain). In addition, tolerance, constipation and dependence were investigated in vivo using experimental paradigms. KEY RESULTS: In mice, pc-morphine was able to elicit antinociceptive effects, both using external light-irradiation (hind paw) and spinal cord implanted fibre-optics. In addition, remote morphine photoactivation was devoid of common systemic opioid-related undesired effects, namely, constipation, tolerance to the analgesic effects, rewarding effects and naloxone-induced withdrawal. CONCLUSION AND IMPLICATIONS: Light-dependent opioid-based drugs may allow effective analgesia without the occurrence of tolerance or the associated and severe opioid-related undesired effects. LINKED ARTICLES: This article is part of a themed issue on Advances in Opioid Pharmacology at the Time of the Opioid Epidemic. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v180.7/issuetoc.
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Analgesia , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos , Ratones , Animales , Morfina/farmacología , Analgésicos Opioides/farmacología , Dolor/tratamiento farmacológico , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/tratamiento farmacológico , Estreñimiento/inducido químicamente , Estreñimiento/tratamiento farmacológicoRESUMEN
En los últimos años se han producido importantes cambios legislativos en numerosos países respecto al consumo de cannabis con fines medicinales y/o recreativos, que han facilitado su accesibilidad. Actualmente, Uruguay, Canadá y algunos estados de EE.UU. han legalizado el consumo recreativo, aplicando distintos modelos legislativos. El objetivo de la presente revisión es analizar los efectos que ha tenido la legalización del cannabis recreativo sobre su consumo y sus consecuencias. En general, las evidencias indican que la legalización se ha asociado a un descenso en el precio, mayor concentración de THC (potencia), mayor diversidad de presentaciones para su consumo, una menor percepción de riesgo y un incremento en el consumo en adultos y de forma moderada en adolescentes (aunque sea ilegal el consumo para ellos), así como un aumento de las consecuencias adversas derivadas del consumo en la salud pública. Se ha producido un descenso en los arrestos relacionados con el consumo, pero el mercado ilegal sigue utilizándose de forma habitual. No se ha detectado un incremento de la demanda de tratamiento por este consumo. Por el momento, estos cambios legislativos no han conseguido alcanzar sus objetivos principales que eran suprimir el mercado ilegal y proteger a los grupos más vulnerables, mientras que, por el contrario, parecen implicar un incremento de algunos aspectos negativos asociados al consumo de cannabis. Sin embargo, teniendo en cuenta que la mayoría de estos cambios legislativos han entrado en vigor hace relativamente poco tiempo, se requiere un periodo de seguimiento mayor para poder extraer conclusiones definitivas. (AU)
In recent years, there have been important legislative changes in many countries regarding the use of cannabis for medicinal and/or recreational purposes, which have facilitated access to it. Uruguay, Canada and some of the US states are the only jurisdictions that have legalised recreational consumption, applying different legislative models. The aim of this review is to analyse the effects that the legalisation of recreational cannabis has had on its use and its consequences. In general, the evidence accumulated to date indicates that the legalisation of cannabis has been associated with a decrease in the price of the substance, higher concentration of THC (potency), greater diversity of presentations for consumption, lower risk perception and an increase in consumption in adults and moderately in adolescents (even though it is illegal for them to consume), as well as an increase in the adverse consequences derived from cannabis consumption on public health. There has been a decrease in drug-related arrests, but the illegal market continues to be frequently used. No increase in the demand for treatment due to cannabis consumption has been detected. Therefore, these legislative changes have so far failed to achieve their main objectives, which were to suppress the illegal market and protect the most vulnerable groups, while on the contrary, they seem to imply an increase in some of the negative aspects associated with cannabis consumption. However, taking into account that most of these legislative changes have entered into force relatively recently, a longer follow-up period is required to be able to draw definitive conclusions. (AU)
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Humanos , Cannabis/crecimiento & desarrollo , Uso de la Marihuana/historia , Uso de la Marihuana/legislación & jurisprudencia , Uso de la Marihuana/tendencias , Salud Pública/legislación & jurisprudencia , Salud Pública/estadística & datos numéricosRESUMEN
Alzheimer's disease (AD) is characterized by a reorganization of brain activity determining network hyperexcitability and loss of synaptic plasticity. Precisely, a dysfunction in metabotropic GABAB receptor signalling through G protein-gated inwardly rectifying K+ (GIRK or Kir3) channels on the hippocampus has been postulated. Thus, we determined the impact of amyloid-ß (Aß) pathology in GIRK channel density, subcellular distribution, and its association with GABAB receptors in hippocampal CA1 pyramidal neurons from the APP/PS1 mouse model using quantitative SDS-digested freeze-fracture replica labelling (SDS-FRL) and proximity ligation in situ assay (P-LISA). In wild type mice, single SDS-FRL detection revealed a similar dendritic gradient for GIRK1 and GIRK2 in CA1 pyramidal cells, with higher densities in spines, and GIRK3 showed a lower and uniform distribution. Double SDS-FRL showed a co-clustering of GIRK2 and GIRK1 in post- and presynaptic compartments, but not for GIRK2 and GIRK3. Likewise, double GABAB1 and GIRK2 SDS-FRL detection displayed a high degree of co-clustering in nanodomains (40-50 nm) mostly in spines and axon terminals. In APP/PS1 mice, the density of GIRK2 and GIRK1, but not for GIRK3, was significantly reduced along the neuronal surface of CA1 pyramidal cells and in axon terminals contacting them. Importantly, GABAB1 and GIRK2 co-clustering was not present in APP/PS1 mice. Similarly, P-LISA experiments revealed a significant reduction in GABAB1 and GIRK2 interaction on the hippocampus of this animal model. Overall, our results provide compelling evidence showing a significant reduction on the cell surface density of pre- and postsynaptic GIRK1 and GIRK2, but not GIRK3, and a decline in GABAB receptors and GIRK2 channels co-clustering in hippocampal pyramidal neurons from APP/PS1 mice, thus suggesting that a disruption in the GABAB receptor-GIRK channel membrane assembly causes dysregulation in the GABAB signalling via GIRK channels in this AD animal model.
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Canales de Potasio Rectificados Internamente Asociados a la Proteína G , Receptores de GABA-B , Animales , Canales de Potasio Rectificados Internamente Asociados a la Proteína G/metabolismo , Canales de Potasio Rectificados Internamente Asociados a la Proteína G/ultraestructura , Hipocampo/metabolismo , Ratones , Plasticidad Neuronal , Receptores de GABA-B/metabolismo , Ácido gamma-AminobutíricoRESUMEN
Aberrant endocannabinoid signaling accompanies several neurodegenerative disorders, including multiple sclerosis. Here, we report altered endocannabinoid signaling in X-linked adrenoleukodystrophy (X-ALD), a rare neurometabolic demyelinating syndrome caused by malfunction of the peroxisomal ABCD1 transporter, resulting in the accumulation of very long-chain fatty acids (VLCFAs). We found abnormal levels of cannabinoid receptor 2 (CB2r) and related endocannabinoid enzymes in the brain and peripheral blood mononuclear cells (PBMCs) of X-ALD patients and in the spinal cord of a murine model of X-ALD. Preclinical treatment with a selective agonist of CB2r (JWH133) halted axonal degeneration and associated locomotor deficits, along with normalization of microgliosis. Moreover, the drug improved the main metabolic disturbances underlying this model, particularly in redox and lipid homeostatic pathways, including increased lipid droplets in motor neurons, through the modulation of the GSK-3ß/NRF2 axis. JWH133 inhibited Reactive Oxygen Species elicited by excess VLCFAs in primary microglial cultures of Abcd1-null mice. Furthermore, we uncovered intertwined redox and CB2r signaling in the murine spinal cords and in patient PBMC samples obtained from a phase II clinical trial with antioxidants (NCT01495260). These findings highlight CB2r signaling as a potential therapeutic target for X-ALD and perhaps other neurodegenerative disorders that present with dysregulated redox and lipid homeostasis.
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Adrenoleucodistrofia , Miembro 1 de la Subfamilia D de Transportador de Casetes de Unión al ATP , Adrenoleucodistrofia/tratamiento farmacológico , Animales , Ensayos Clínicos Fase II como Asunto , Endocannabinoides/uso terapéutico , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Leucocitos Mononucleares/metabolismo , Ratones , Factor 2 Relacionado con NF-E2/metabolismo , Receptores de Cannabinoides/metabolismo , Receptores de Cannabinoides/uso terapéuticoRESUMEN
G protein-coupled receptors (GPCRs) constitute the largest group of membrane receptor proteins controlling brain activity. Accordingly, GPCRs are the main target of commercial drugs for most neurological and neuropsychiatric disorders. One of the mechanisms by which GPCRs regulate neuronal function is by homo- and heteromerization, with the establishment of direct protein-protein interactions between the same and different GPCRs. The occurrence of GPCR homo- and heteromers in artificial systems is generally well accepted, but more specific methods are necessary to address GPCR oligomerization in the brain. Here, we revise some of the techniques that have mostly contributed to reveal GPCR oligomers in native tissue, which include immunogold electron microscopy, proximity ligation assay (PLA), resonance energy transfer (RET) between fluorescent ligands and the Amplified Luminescent Proximity Homogeneous Assay (ALPHA). Of note, we use the archetypical GPCR oligomer, the adenosine A2A receptor (A2AR)-dopamine D2 receptor (D2R) heteromer as an example to illustrate the implementation of these techniques, which can allow visualizing GPCR oligomers in the human brain under normal and pathological conditions. Indeed, GPCR oligomerization may be involved in the pathophysiology of neurological and neuropsychiatric disorders.
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Receptores Dopaminérgicos , Receptores Acoplados a Proteínas G , Adenosina , Encéfalo/metabolismo , Humanos , Ligandos , Receptores Dopaminérgicos/metabolismo , Receptores Acoplados a Proteínas G/metabolismoRESUMEN
Amyloid plaques composed of Aß amyloid peptides and neurofibrillary tangles are a pathological hallmark of Alzheimer Disease. In situ identification of early-stage amyloid aggregates in Alzheimer's disease is relevant for their importance as potential targets for effective drugs. Synchrotron-based infrared imaging is here used to identify early-stage oligomeric/granular aggregated amyloid species in situ in the brain of APP/PS1 transgenic mice for the first time. Also, APP/PS1 mice show fibrillary aggregates at 6 and 12 months. A significant decreased burden of early-stage aggregates and fibrillary aggregates is obtained following treatment with poly(propylene imine) dendrimers with histidine-maltose shell (a neurodegenerative protector) in 6-month-old APP/PS1 mice, thus demonstrating their putative therapeutic properties of in AD models. Identification, localization, and characterization using infrared imaging of these non-fibrillary species in the cerebral cortex at early stages of AD progression in transgenic mice point to their relevance as putative pharmacological targets. No less important, early detection of these structures may be useful in the search for markers for non-invasive diagnostic techniques.
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Enfermedad de Alzheimer/tratamiento farmacológico , Dendrímeros/uso terapéutico , Polipropilenos/uso terapéutico , Péptidos beta-Amiloides/genética , Péptidos beta-Amiloides/metabolismo , Animales , Corteza Cerebral/diagnóstico por imagen , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Dendrímeros/administración & dosificación , Histidina/química , Maltosa/química , Ratones , Ratones Endogámicos C57BL , Polipropilenos/administración & dosificación , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Amyloid plaques are one of the principal hallmarks of Alzheimer's disease and are mainly composed of Aß amyloid peptides together with other components such as lipids, cations, or glycosaminoglycans. The structure of amyloid peptide's aggregates is related to the peptide toxicity and highly depends on the aggregation conditions and the presence of cofactors. While fibrillary aggregates are nowadays considered nontoxic, oligomeric/granular (nonfibrillary) aggregates have been found to be toxic. In this work we have characterized in situ two different types of amyloid deposits analyzing sections of the cortex of patients in advanced stages of Alzheimer disease. By combining SR-µFTIR for the study of the secondary structure of the peptide and ThS fluorescence as an indicator of fibrillary structures, we found two types of plaques: ThS positive plaques with a clear infrared band at 1630 cm-1 that would correspond to fibrillary plaques and ThS negative plaques showing a mixture of nonfibrillar ß-sheet and unordered aggregated structures that would correspond to the nonfibrillary plaques (plaques with increased unordered structure). The analysis of the FTIR spectra has allowed correlation of lipid oxidation with the presence of nonfibrillary plaques. The metal composition of the two types of plaques has been analyzed using SR-nano-XRF and XANES. The results have shown higher accumulation of iron (mainly Fe2+) in fibrillary plaques than in nonfibrillary ones. However, in nonfibrillary plaques Fe3+ has been found to predominate over Fe2+. The identification of different types of aggregated forms and the different composition of metals found in the different types of plaques could be of paramount importance for the understanding of the development of Alzheimer disease.
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Enfermedad de Alzheimer , Placa Amiloide , Péptidos beta-Amiloides , Humanos , Espectroscopía Infrarroja por Transformada de Fourier , Sincrotrones , Rayos XRESUMEN
According to the adenosine hypothesis of schizophrenia, the classically associated hyperdopaminergic state may be secondary to a loss of function of the adenosinergic system. Such a hypoadenosinergic state might either be due to a reduction of the extracellular levels of adenosine or alterations in the density of adenosine A2A receptors (A2ARs) or their degree of functional heteromerization with dopamine D2 receptors (D2R). In the present study, we provide preclinical and clinical evidences for this latter mechanism. Two animal models for the study of schizophrenia endophenotypes, namely the phencyclidine (PCP) mouse model and the A2AR knockout mice, were used to establish correlations between behavioural and molecular studies. In addition, a new AlphaLISA-based method was implemented to detect native A2AR-D2R heteromers in mouse and human brain. First, we observed a reduction of prepulse inhibition in A2AR knockout mice, similar to that observed in the PCP animal model of sensory gating impairment of schizophrenia, as well as a significant upregulation of striatal D2R without changes in A2AR expression in PCP-treated animals. In addition, PCP-treated animals showed a significant reduction of striatal A2AR-D2R heteromers, as demonstrated by the AlphaLISA-based method. A significant and pronounced reduction of A2AR-D2R heteromers was next demonstrated in postmortem caudate nucleus from schizophrenic subjects, even though both D2R and A2AR were upregulated. Finally, in PCP-treated animals, sub-chronic administration of haloperidol or clozapine counteracted the reduction of striatal A2AR-D2R heteromers. The degree of A2AR-D2R heteromer formation in schizophrenia might constitute a hallmark of the illness, which indeed should be further studied to establish possible correlations with chronic antipsychotic treatments.
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Receptor de Adenosina A2A , Esquizofrenia , Adenosina , Animales , Cuerpo Estriado/metabolismo , Dopamina , Ratones , Receptor de Adenosina A2A/genética , Receptor de Adenosina A2A/metabolismo , Receptores de Dopamina D2/metabolismoRESUMEN
Editor's Note: this Article has been retracted; the Retraction Note is available at https://www.nature.com/articles/s41598-020-76208-w.
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Introduction: Lafora disease (LD) is a rare form of progressive infantile epilepsy in which rapid neurological deterioration occurs as the disease advances, leading the patients to a vegetative state and then death, usually within the first decade of disease onset. Based on the capacity of the endogenous cannabinoid system (ECS) to modulate several cellular processes commonly altered in many neurodegenerative processes, as well as the antiepileptic properties of certain natural cannabinoids, the aim of this study was to evaluate the role of the ECS in LD progression. Materials and Methods: We tested whether a natural cannabis extract highly enriched in cannabidiol (CBD) might be effective in curbing the pathological phenotype of malin knockout (KO) mice as an animal model of LD. Results: Our results reveal for the first time that alterations in the ECS occur during the evolution of LD, mainly at the level of CB1, CB2, and G protein-coupled receptor 55 (GPR55) receptor expression, and that a CBD-enriched extract (CBDext) is able to reduce the cognitive impairment exhibited by malin KO mice. However, in contrast to what has previously been reported for other kinds of refractory epilepsy in childhood, the CBD-enriched extract does not reduce the severity of the epileptic seizures induced in this animal model of LD. Conclusions: In summary, this study reveals that the ECS might play a role in LD and that a CBD-enriched extract partially reduces the dementia-like phenotype, but not the increased vulnerability to epileptic seizures, exhibited by an animal model of such a life-threatening disease.
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Oxidative stress is a major pathogenic factor in Alzheimer's disease, but it should not be tackled alone rather together with other key targets to derive effective treatments. The combination of the scaffold of the polar antioxidant lead 7-methoxy-2,2-dimethylchroman-6-ol (CR-6) with that of the lipophilic cholinesterase inhibitor 6-chlorotacrine results in compounds with favorable brain permeability and multiple activities in vitro (acetylcholinesterase, butyrylcholinesterase, ß-site amyloid precursor protein (APP) cleaving enzyme-1 (BACE-1), and Aß42 and tau aggregation inhibition). In in vivo studies on wild-type and APP/presenilin 1 (PS1) mice, two selected compounds were well tolerated and led to positive trends, albeit statistically nonsignificant in some cases, on memory performance, amyloid pathology (reduced amyloid burden and potentiated non-amyloidogenic APP processing), and oxidative stress (reduced cortical oxidized proteins and increased antioxidant enzymes superoxide dismutase 2 (SOD2), catalase, glutathione peroxidase 1 (GPX1), and heme oxygenase 1 (Hmox1) and transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf2)). These compounds emerge as interesting brain-permeable multitarget compounds, with a potential as anti-Alzheimer agents beyond that of the original lead CR-6.
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Enfermedad de Alzheimer/tratamiento farmacológico , Antioxidantes/química , Antioxidantes/farmacología , Benzopiranos/química , Benzopiranos/farmacología , Terapia Molecular Dirigida , Enfermedad de Alzheimer/metabolismo , Animales , Antioxidantes/metabolismo , Antioxidantes/uso terapéutico , Benzopiranos/metabolismo , Benzopiranos/uso terapéutico , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Humanos , Ratones , Simulación de Dinámica Molecular , Estrés Oxidativo/efectos de los fármacos , Permeabilidad , Conformación ProteicaRESUMEN
Amyloid plaques composed of Aß amyloid peptides and neurofibrillary tangles are a pathological hallmark of Alzheimer's disease. In situ identification of early-stage amyloid aggregates in Alzheimer's disease is relevant for their importance as potential targets for effective drugs. Synchrotron-based infrared imaging is here used to identify early-stage oligomeric/granular aggregated amyloid species in situ in the brain of APP/PS1 transgenic mice and Octodon degus for the first time. Also, APP/PS1 mice show fibrillary aggregates at 6 and 12 months whereas very little formation of fibrils is found in aged Octodon degus. Finally, significant decreased burden of early-stage aggregates and fibrillary aggregates is obtained following treatment with G4-His-Mal dendrimers (a neurodegenerative protector) in 6-month-old APP/PS1 mice, thus demonstrating putative therapeutic properties of G4-His-Mal dendrimers in AD models. Identification, localization, and characterization using infrared imaging of these non-fibrillary species in the cerebral cortex at early stages of AD progression in transgenic mice point to their relevance as putative pharmacological targets. No less important, early detection of these structures may be useful in the search for markers for non-invasive diagnostic techniques.
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Enfermedad de Alzheimer/patología , Placa Amiloide/patología , Factores de Edad , Enfermedad de Alzheimer/diagnóstico , Animales , Encéfalo/patología , Modelos Animales de Enfermedad , Masculino , Ratones , Ratones Transgénicos , Octodon , Espectroscopía Infrarroja por Transformada de Fourier , SincrotronesRESUMEN
The deposition of aggregated amyloid-ß peptides derived from the pro-amyloidogenic processing of the amyloid precurson protein (APP) into characteristic amyloid plaques (APs) is distinctive to Alzheimer's disease (AD). Alternative APP processing via the metalloprotease ADAM10 prevents amyloid-ß formation. We tested whether downregulation of ADAM10 activity by its secreted endogenous inhibitor secreted-frizzled-related protein 1 (SFRP1) is a common trait of sporadic AD. We demonstrate that SFRP1 is significantly increased in the brain and cerebrospinal fluid of patients with AD, accumulates in APs and binds to amyloid-ß, hindering amyloid-ß protofibril formation. Sfrp1 overexpression in an AD-like mouse model anticipates the appearance of APs and dystrophic neurites, whereas its genetic inactivation or the infusion of α-SFRP1-neutralizing antibodies favors non-amyloidogenic APP processing. Decreased Sfrp1 function lowers AP accumulation, improves AD-related histopathological traits and prevents long-term potentiation loss and cognitive deficits. Our study unveils SFRP1 as a crucial player in AD pathogenesis and a promising AD therapeutic target.
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Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/metabolismo , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Proteína ADAM10/biosíntesis , Proteína ADAM10/genética , Enfermedad de Alzheimer/patología , Secretasas de la Proteína Precursora del Amiloide/biosíntesis , Secretasas de la Proteína Precursora del Amiloide/genética , Precursor de Proteína beta-Amiloide/genética , Animales , Anticuerpos Bloqueadores/uso terapéutico , Química Encefálica/genética , Regulación hacia Abajo , Humanos , Potenciación a Largo Plazo , Proteínas de la Membrana/antagonistas & inhibidores , Proteínas de la Membrana/biosíntesis , Ratones , Ratones Transgénicos , Neuritas/patología , Placa Amiloide/tratamiento farmacológico , Placa Amiloide/genética , Placa Amiloide/patologíaRESUMEN
Poly(propylene imine) dendrimers have been shown to be promising 3-dimensional polymers for the use in the pharmaceutical and biomedical applications. Our aims of this study were first, to synthesize a novel type of dendrimer with poly(propylene imine) core and maltose-histidine shell (G4HisMal) assessing if maltose-histidine shell can improve the biocompatibility and the ability to cross the blood-brain barrier, and second, to investigate the potential of G4HisMal to protect Alzheimer disease transgenic mice from memory impairment. Our data demonstrate that G4HisMal has significantly improved biocompatibility and ability to cross the blood-brain barrier in vivo. Therefore, we suggest that a maltose-histidine shell can be used to improve biocompatibility and ability to cross the blood-brain barrier of dendrimers. Moreover, G4HisMal demonstrated properties for synapse and memory protection when administered to Alzheimer disease transgenic mice. Therefore, G4HisMal can be considered as a promising drug candidate to prevent Alzheimer disease via synapse protection.
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Histidina/uso terapéutico , Maltosa/uso terapéutico , Trastornos de la Memoria/prevención & control , Fármacos Neuroprotectores/uso terapéutico , Polipropilenos/uso terapéutico , Enfermedad de Alzheimer/complicaciones , Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/patología , Animales , Línea Celular , Dendrímeros/química , Dendrímeros/farmacocinética , Dendrímeros/uso terapéutico , Histidina/análogos & derivados , Histidina/farmacocinética , Humanos , Maltosa/análogos & derivados , Maltosa/farmacocinética , Trastornos de la Memoria/complicaciones , Trastornos de la Memoria/patología , Ratones , Ratones Transgénicos , Nanopartículas/química , Nanopartículas/uso terapéutico , Fármacos Neuroprotectores/química , Fármacos Neuroprotectores/farmacocinética , Polipropilenos/química , Polipropilenos/farmacocinética , Sinapsis/efectos de los fármacos , Sinapsis/patologíaRESUMEN
At present, clinical interest in the plant-derived cannabinoid compound cannabidiol (CBD) is rising exponentially, since it displays multiple therapeutic properties. In addition, CBD can counteract the undesirable effects of the psychoactive cannabinoid Δ9-tetrahydrocannabinol (Δ9-THC) that hinder clinical development of cannabis-based therapies. Despite this attention, the mechanisms of CBD action and its interaction with Δ9-THC are still not completely elucidated. Here, by combining in vivo and complementary molecular techniques, we demonstrate for the first time that CBD blunts the Δ9-THC-induced cognitive impairment in an adenosine A2A receptor (A2AR)-dependent manner. Furthermore, we reveal the existence of A2AR and cannabinoid CB1 receptor (CB1R) heteromers at the presynaptic level in CA1 neurons in the hippocampus. Interestingly, our findings support a brain region-dependent A2AR-CB1R functional interplay; indeed, CBD was not capable of modifying motor functions presumably regulated by striatal A2AR/CB1R complexes, nor anxiety responses related to other brain regions. Overall, these data provide new evidence regarding the mechanisms of action of CBD and the nature of A2AR-CB1R interactions in the brain.
Asunto(s)
Cannabidiol/uso terapéutico , Disfunción Cognitiva/inducido químicamente , Disfunción Cognitiva/tratamiento farmacológico , Dronabinol/efectos adversos , Hipocampo/metabolismo , Multimerización de Proteína , Receptor de Adenosina A2A/metabolismo , Receptor Cannabinoide CB1/metabolismo , Animales , Cannabidiol/farmacología , Disfunción Cognitiva/fisiopatología , Hipocampo/fisiopatología , Hipocampo/ultraestructura , Locomoción/efectos de los fármacos , Masculino , Ratones Endogámicos C57BL , Terminales Presinápticos/efectos de los fármacos , Terminales Presinápticos/metabolismo , Multimerización de Proteína/efectos de los fármacosRESUMEN
OBJECTIVE: The first aim of this study was to develop a nanocarrier that could transport the peroxisome proliferator-activated receptor agonist, pioglitazone (PGZ) across brain endothelium and examine the mechanism of nanoparticle transcytosis. The second aim was to determine whether these nanocarriers could successfully treat a mouse model of Alzheimer's disease (AD). METHODS: PGZ-loaded nanoparticles (PGZ-NPs) were synthesized by the solvent displacement technique, following a factorial design using poly (lactic-co-glycolic acid) polyethylene glycol (PLGA-PEG). The transport of the carriers was assessed in vitro, using a human brain endothelial cell line, cytotoxicity assays, fluorescence-tagged nanocarriers, fluorescence-activated cell sorting, confocal and transmission electron microscopy. The effectiveness of the treatment was assessed in APP/PS1 mice in a behavioral assay and by measuring the cortical deposition of ß-amyloid. RESULTS: Incorporation of PGZ into the carriers promoted a 50x greater uptake into brain endothelium compared with the free drug and the carriers showed a delayed release profile of PGZ in vitro. In the doses used, the nanocarriers were not toxic for the endothelial cells, nor did they alter the permeability of the blood-brain barrier model. Electron microscopy indicated that the nanocarriers were transported from the apical to the basal surface of the endothelium by vesicular transcytosis. An efficacy test carried out in APP/PS1 transgenic mice showed a reduction of memory deficit in mice chronically treated with PGZ-NPs. Deposition of ß-amyloid in the cerebral cortex, measured by immunohistochemistry and image analysis, was correspondingly reduced. CONCLUSION: PLGA-PEG nanocarriers cross brain endothelium by transcytosis and can be loaded with a pharmaceutical agent to effectively treat a mouse model of AD.