RESUMEN
Thioredoxin interacting protein (TXNIP) has emerged as a significant regulator of ß-cell mass and loss, rendering it an attractive target for treating diabetes. We previously showed that Shiga-Y6, a fluorinated curcumin derivative, inhibited TXNIP mRNA and protein expression in vitro, raising the question of whether the same effect could be translated in vivo. Herein, we examined the effect of Shiga-Y6 on TNXIP levels and explored its therapeutic potential in a mouse model of diabetes, Akita mice. We intraperitoneally injected Shiga-Y6 (SY6; 30 mg/kg of body weight) or vehicle into 8-week-old Akita mice for 28 consecutive days. On day 29, the mice were euthanized, following which the serum levels of glucose, insulin, and glucagon were measured using ELISA, the expression of TXNIP in pancreatic tissue lysates was determined using western blotting, and the level of ß-cell apoptosis was assessed using the TUNEL assay. TXNIP levels in the pancreatic tissue of Akita mice were significantly elevated compared with wild-type (WT) mice. Shiga-Y6 administration for 28 days significantly lowered those levels compared with Akita mice that received vehicle to a level comparable to WT mice. In immunohistochemical analysis, both α- to ß-cell ratio and the number of apoptotic ß-cells were significantly reduced in SY6-treated Akita mice, compared with vehicle-treated Akita mice. Findings from the present study suggest a potential of Shiga-Y6 as an antidiabetic agent through lowering TXNIP protein levels and ameliorating pancreatic ß-cells apoptosis.
Asunto(s)
Curcumina , Diabetes Mellitus , Células Secretoras de Insulina , Ratones , Animales , Curcumina/farmacología , Curcumina/uso terapéutico , Curcumina/metabolismo , Diabetes Mellitus/metabolismo , Células Secretoras de Insulina/metabolismo , Insulina/metabolismo , Modelos Animales de Enfermedad , Tiorredoxinas/genética , Tiorredoxinas/metabolismoRESUMEN
Mitochondrial ferritin (FtMt) is a mitochondrial iron storage protein associated with neurodegenerative diseases. In patients with progressive supranuclear palsy (PSP), FtMt was shown to accumulate in nigral neurons. Here, we investigated FtMt and LC3 in the post-mortem midbrain of PSP patients to reveal novel aspects of the pathology. Immunohistochemistry was used to assess the distribution and abnormal changes in FtMt and LC3 immunoreactivities. Colocalization analysis using double immunofluorescence was performed, and subcellular patterns were examined using 3D imaging and modeling. In the substantia nigra pars compacta (SNc), strong FtMt-IR and LC3-IR were observed in the neurons of PSP patients. In other midbrain regions, such as the superior colliculus, the FtMt-IR and LC3-IR remained unchanged. In the SNc, nigral neurons were categorized into four patterns based on subcellular LC3/FtMt immunofluorescence intensities, degree of colocalization, and subcellular overlapping. This categorization suggested that concomitant accumulation of LC3/FtMt is related to mitophagy processes. Using the LC3-IR to stage neuronal damage, we retraced LC3/FtMt patterns and revealed the progression of FtMt accumulation in nigral neurons. Informed by these findings, we proposed a hypothesis to explain the function of FtMt during PSP progression.
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Ferritinas/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismo , Proteínas Mitocondriales/metabolismo , Neuronas/metabolismo , Sustancia Negra/metabolismo , Parálisis Supranuclear Progresiva/metabolismo , Biomarcadores , Susceptibilidad a Enfermedades , Ferritinas/genética , Técnica del Anticuerpo Fluorescente , Humanos , Inmunohistoquímica , Mesencéfalo/metabolismo , Mesencéfalo/patología , Proteínas Asociadas a Microtúbulos/genética , Mitocondrias/metabolismo , Proteínas Mitocondriales/genética , Mitofagia , Unión Proteica , Transporte de Proteínas , Sustancia Negra/patología , Parálisis Supranuclear Progresiva/diagnóstico , Parálisis Supranuclear Progresiva/etiologíaRESUMEN
Amyloid-ß (Aß) accumulation and tauopathy are considered the pathological hallmarks of Alzheimer's disease (AD), but attenuation in choline signaling, including decreased nicotinic acetylcholine receptors (nAChRs), is evident in the early phase of AD. Currently, there are no drugs that can suppress the progression of AD due to a limited understanding of AD pathophysiology. For this, diagnostic methods that can assess disease progression non-invasively before the onset of AD symptoms are essential, and it would be valuable to incorporate the concept of neurotheranostics, which simultaneously enables diagnosis and treatment. The neuroprotective pathways activated by nAChRs are attractive targets as these receptors may regulate microglial-mediated neuroinflammation. Microglia exhibit both pro- and anti-inflammatory functions that could be modulated to mitigate AD pathogenesis. Currently, single-cell analysis is identifying microglial subpopulations that may have specific functions in different stages of AD pathologies. Thus, the ability to image nAChRs and microglia in AD according to the stage of the disease in the living brain may lead to the development of new diagnostic and therapeutic methods. In this review, we summarize and discuss the recent findings on the nAChRs and microglia, as well as their methods for live imaging in the context of diagnosis, prophylaxis, and therapy for AD.
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Enfermedad de Alzheimer , Receptores Nicotínicos , Enfermedad de Alzheimer/diagnóstico por imagen , Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Encéfalo/metabolismo , Humanos , Microglía/metabolismo , Receptores Nicotínicos/metabolismoRESUMEN
Thioredoxin-interacting protein (TXNIP) is involved in multiple disease-associated functions related to oxidative stress, especially by inhibiting the anti-oxidant- and thiol-reducing activity of thioredoxin (TXN). Shiga-Y5 (SY5), a fluorine-19 magnetic resonance probe for detecting amyloid-ß deposition in the brain, previously showed therapeutic effects in a mouse model of Alzheimer's disease; however, the mechanism of action of SY5 remains unclear. SY5 passes the blood-brain barrier and then undergoes hydrolysis to produce a derivative, Shiga-Y6 (SY6), which is a TXNIP-negative regulator. Therefore, this study investigates the therapeutic role of SY5 as the prodrug of SY6 in the thioredoxin system in the brain of a mouse model of Alzheimer's disease. The intraperitoneal injection of SY5 significantly inhibited TXNIP mRNA (p = 0.0072) and protein expression (p = 0.0143) induced in the brain of APP/PS1 mice. In contrast, the levels of TXN mRNA (p = 0.0285) and protein (p = 0.0039) in the brain of APP/PS1 mice were increased after the injection of SY5. The ratio of TXN to TXNIP, which was decreased (p = 0.0131) in the brain of APP/PS1 mice, was significantly increased (p = 0.0072) after the injection of SY5. These results suggest that SY5 acts as a prodrug of SY6 in targeting the thioredoxin system and could be a potential therapeutic compound in oxidative stress-related diseases in the brain.
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Enfermedad de Alzheimer/tratamiento farmacológico , Encéfalo/efectos de los fármacos , Proteínas Portadoras/antagonistas & inhibidores , Curcumina/farmacología , Modelos Animales de Enfermedad , Sondas Moleculares/farmacología , Enfermedad de Alzheimer/metabolismo , Animales , Encéfalo/metabolismo , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Curcumina/administración & dosificación , Curcumina/análogos & derivados , Flúor , Inyecciones Intraperitoneales , Imagen por Resonancia Magnética , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Sondas Moleculares/administración & dosificación , Sondas Moleculares/química , Estructura MolecularRESUMEN
Recent evidence suggests that the formation of soluble amyloid ß (Aß) aggregates with high toxicity, such as oligomers and protofibrils, is a key event that causes Alzheimer's disease (AD). However, understanding the pathophysiological role of such soluble Aß aggregates in the brain in vivo could be difficult due to the lack of a clinically available method to detect, visualize, and quantify soluble Aß aggregates in the brain. We had synthesized a novel fluorinated curcumin derivative with a fixed keto form, named as Shiga-Y51, which exhibited high selectivity to Aß oligomers in vitro. In this study, we investigated the in vivo detection of Aß oligomers by fluorine-19 (19F) magnetic resonance imaging (MRI) using Shiga-Y51 in an APP/PS1 double transgenic mouse model of AD. Significantly high levels of 19F signals were detected in the upper forebrain region of APP/PS1 mice compared with wild-type mice. Moreover, the highest levels of Aß oligomers were detected in the upper forebrain region of APP/PS1 mice in enzyme-linked immunosorbent assay. These findings suggested that 19F-MRI using Shiga-Y51 detected Aß oligomers in the in vivo brain. Therefore, 19F-MRI using Shiga-Y51 with a 7 T MR scanner could be a powerful tool for imaging Aß oligomers in the brain.
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Enfermedad de Alzheimer/patología , Péptidos beta-Amiloides/metabolismo , Encéfalo/patología , Curcumina/metabolismo , Modelos Animales de Enfermedad , Imagen por Resonancia Magnética con Fluor-19/métodos , Placa Amiloide/patología , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/química , Animales , Encéfalo/metabolismo , Curcumina/química , Femenino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Placa Amiloide/metabolismoRESUMEN
Thioredoxin-interacting protein (TXNIP) has multiple disease-associated functions including inducing oxidative stress by inhibiting the anti-oxidant and thiol reducing activity of thioredoxin (TRX), reducing cellular glucose transport, and is a component of the activated inflammasome complex. Increased expression of TXNIP is encountered in diabetic conditions of high glucose. Curcumin and chemical derivatives have multiple therapeutic properties as anti-inflammatories, anti-oxidants, amyloid aggregation inhibitors and modulate a number of cellular signaling pathways. Using a fluorinated-derivative of curcumin (designated Shiga-Y6), we showed significant inhibition of TXNIP mRNA and protein expression, and induction of TRX mRNA and protein in ARPE-19 retinal pigment epithelial cells and THP-1-derived macrophages, while the non-fluorinated structural equivalent (Shiga-Y52) and native curcumin did not show these same effects. Shiga-Y6 was effective in reducing high glucose, endoplasmic reticulum stress-induced TXNIP in ARPE-19 cells, and reducing lipopolysaccharide and endoplasmic stress-induced proinflammatory gene expression in THP-1 macrophages. Moreover, TXNIP-knockdown experiments showed that the anti-inflammatory effect of Shiga-Y6 in LPS-stimulated THP-1 macrophages was TXNIP-independent.
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Antiinflamatorios/farmacología , Proteínas Portadoras/metabolismo , Macrófagos/efectos de los fármacos , Epitelio Pigmentado de la Retina/efectos de los fármacos , Antiinflamatorios/química , Proteínas Portadoras/genética , Línea Celular , Curcumina/farmacología , Estrés del Retículo Endoplásmico , Técnicas de Silenciamiento del Gen , Glucosa/farmacología , Humanos , Mediadores de Inflamación/metabolismo , Macrófagos/metabolismo , Epitelio Pigmentado de la Retina/metabolismo , Células THP-1 , Tiorredoxinas/metabolismoRESUMEN
Iron deposits are often observed in the brains of patients with neurodegenerative diseases, including Alzheimer's and Parkinson's diseases. This study outlines the development of F-Nox-1 as the first example of a 19F-MRI probe that can selectively detect Fe(ii) in aqueous solutions. The use of tetrafluoro-p-phenylenediamine (TFPDA) as a 19F signal emitter with an Fe(ii)-selective chemical switch, based on our previously reported N-oxide chemistry, yielded a readout of a symmetry-dependent 19F signal change in response to Fe(ii). The addition of Fe(ii) ions to F-Nox-1 triggered a 19F signal change, both in the chemical shift and signal intensity, and the response was highly selective to Fe(ii) over other biologically relevant metal ions. The probe could also detect Fe(ii) in serum containing various biological contaminants by 19F magnetic resonance imaging (19F-MRI). Imaging of soluble Fe(ii) species, which is the major component of water-soluble iron species, by 19F-MRI will potentially enable the direct monitoring of the elevation of Fe(ii) levels prior to the formation of iron deposits, which is a potential risk factor for neurodegenerative diseases.
RESUMEN
Oxidative stress is associated with the progression of the neurodegenerative diseases Parkinson's disease (PD) and cerebral ischemia. Recently, 5-aminolevulinic acid (5-ALA), an intermediate in the porphyrin synthesis pathway, was reported to exert antioxidative effects on macrophages and cardiomyocytes. Here, we demonstrated the neuroprotective effects of 5-ALA using rat models of PD and ischemia as well as in vitro in SH-SY5Y cells. 5-ALA partially prevented neurodegeneration in each condition. These results suggest that 5-ALA has a potential for promising therapeutic agent to protect against neurodegeneration exacerbated by oxidative stress.
Asunto(s)
Isquemia Encefálica/patología , Ácidos Levulínicos/farmacología , Degeneración Nerviosa , Fármacos Neuroprotectores , Estrés Oxidativo/efectos de los fármacos , Enfermedad de Parkinson/patología , Accidente Cerebrovascular/patología , Animales , Isquemia Encefálica/etiología , Línea Celular Tumoral , Modelos Animales de Enfermedad , Humanos , Ácidos Levulínicos/uso terapéutico , Masculino , Degeneración Nerviosa/prevención & control , Enfermedad de Parkinson/etiología , Ratas Wistar , Accidente Cerebrovascular/etiología , Ácido AminolevulínicoRESUMEN
Aberrant angiogenesis is a pathological feature of a number of diseases and arises from the uncoordinated expression of angiogenic factors as response to different cellular stresses. Age-related macular degeneration (AMD), a leading cause of vision loss, can result from pathological angiogenesis. As a mutation in the mitochondrial ferritin (FTMT) gene has been associated with AMD, its possible role in modulating angiogenic factors and angiogenesis was investigated. FTMT is an iron-sequestering protein primarily expressed in metabolically active cells and tissues with high oxygen demand, including retina. In this study, we utilized the human retinal pigment epithelial cell line ARPE-19, both as undifferentiated and differentiated cells. The effects of proinflammatory cytokines, FTMT knockdown, and transient and stable overexpression of FTMT were investigated on expression of pro-angiogenic vascular endothelial growth factor (VEGF) and anti-angiogenic pigment epithelial-derived factor (PEDF). Proinflammatory cytokines induced FTMT and VEGF expression, while NF-κB inhibition significantly reduced FTMT expression. VEGF protein and mRNA expression were significantly increased in FTMT-silenced ARPE-19 cells. Using an in vitro angiogenesis assay with endothelial cells, we showed that conditioned media from FTMT-overexpressing cells had significant antiangiogenic effects. Collectively, our findings indicate that increased levels of FTMT inhibit angiogenesis, possibly by reducing levels of VEGF and increasing PEDF expression. The cellular models developed can be used to investigate if increased FTMT may be protective in angiogenic diseases, such as AMD.
Asunto(s)
Ferritinas/metabolismo , Proteínas Mitocondriales/metabolismo , Neovascularización Fisiológica , Epitelio Pigmentado de la Retina/metabolismo , Línea Celular , Citocinas/genética , Citocinas/metabolismo , Proteínas del Ojo/genética , Proteínas del Ojo/metabolismo , Ferritinas/genética , Humanos , Proteínas Mitocondriales/genética , FN-kappa B/genética , FN-kappa B/metabolismo , Factores de Crecimiento Nervioso/genética , Factores de Crecimiento Nervioso/metabolismo , Epitelio Pigmentado de la Retina/citología , Serpinas/genética , Serpinas/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Aggregation of tau into neurofibrillary tangles (NFTs) is characteristic of tauopathies, including Alzheimer's disease. Recent advances in tau imaging have attracted much attention because of its potential contributions to early diagnosis and monitoring of disease progress. Fluorine-19 magnetic resonance imaging (19 F-MRI) may be extremely useful for tau imaging once a high-quality probe has been formulated. In this investigation, a novel fluorine-19-labeling compound has been developed as a probe for tau imaging using 19 F-MRI. This compound is a buta-1,3-diene derivative with a polyethylene glycol side chain bearing a CF3 group and is known as Shiga-X35. Female rTg4510 mice (a mouse model of tauopathy) and wild-type mice were intravenously injected with Shiga-X35, and magnetic resonance imaging of each mouse's head was conducted in a 7.0-T horizontal-bore magnetic resonance scanner. The 19 F-MRI in rTg4510 mice showed an intense signal in the forebrain region. Analysis of the signal intensity in the forebrain region revealed a significant accumulation of fluorine-19 magnetic resonance signal in the rTg4510 mice compared with the wild-type mice. Histological analysis showed fluorescent signals of Shiga-X35 binding to the NFTs in the brain sections of rTg4510 mice. Data collected as part of this investigation indicate that 19 F-MRI using Shiga-X35 could be a promising tool to evaluate tau pathology in the brain.
Asunto(s)
Benzoxazoles/química , Butadienos/química , Imagen por Resonancia Magnética con Fluor-19/métodos , Flúor , Tauopatías/diagnóstico por imagen , Anciano , Enfermedad de Alzheimer/diagnóstico por imagen , Enfermedad de Alzheimer/metabolismo , Animales , Benzoxazoles/síntesis química , Encéfalo/diagnóstico por imagen , Encéfalo/metabolismo , Butadienos/síntesis química , Modelos Animales de Enfermedad , Femenino , Humanos , Masculino , Ratones , Ratones Transgénicos , Ovillos Neurofibrilares/metabolismo , Proteínas tau/metabolismoRESUMEN
Charcot-Marie-Tooth disease (CMT) is the most common inherited neuropathy characterized by clinical and genetic heterogeneity. Although more than 30 loci harboring CMT-causing mutations have been identified, many other genes still remain to be discovered for many affected individuals. For two consanguineous families with CMT (axonal and mixed phenotypes), a parametric linkage analysis using genome-wide SNP chip identified a 4.3 Mb region on 12q24 showing a maximum multipoint LOD score of 4.23. Subsequent whole-genome sequencing study in one of the probands, followed by mutation screening in the two families, revealed a disease-specific 5 bp deletion (c.247-10_247-6delCACTC) in a splicing element (pyrimidine tract) of intron 2 adjacent to the third exon of cytochrome c oxidase subunit VIa polypeptide 1 (COX6A1), which is a component of mitochondrial respiratory complex IV (cytochrome c oxidase [COX]), within the autozygous linkage region. Functional analysis showed that expression of COX6A1 in peripheral white blood cells from the affected individuals and COX activity in their EB-virus-transformed lymphoblastoid cell lines were significantly reduced. In addition, Cox6a1-null mice showed significantly reduced COX activity and neurogenic muscular atrophy leading to a difficulty in walking. Those data indicated that COX6A1 mutation causes the autosomal-recessive axonal or mixed CMT.
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Axones/fisiología , Enfermedad de Charcot-Marie-Tooth/genética , Complejo IV de Transporte de Electrones/genética , Complejo IV de Transporte de Electrones/fisiología , Genes Recesivos/genética , Atrofia Muscular/genética , Mutación/genética , Adulto , Animales , Consanguinidad , Electrofisiología , Femenino , Ligamiento Genético , Humanos , Escala de Lod , Masculino , Ratones , Ratones Noqueados , Linaje , Fenotipo , Empalme del ARN/genéticaRESUMEN
We have recently shown that age-dependent regional brain atrophy and lateral ventricle expansion may be linked with impaired cognitive and locomotor functions. However, metabolic profile transformation in different brain regions during aging is unknown. This study examined metabolic changes in the hippocampus, medial prefrontal cortex (mPFC) and striatum of middle- and late-aged Sprague-Dawley rats using ultrahigh performance liquid chromatography coupled with high-resolution accurate mass-orbitrap tandem mass spectrometry. Thirty-eight potential metabolites were altered in hippocampus, 29 in mPFC, and 14 in striatum. These alterations indicated that regional metabolic mechanisms in lated-aged rats are related to multiple pathways including glutathione, sphingolipid, tyrosine, and purine metabolism. Thus, our findings might be useful for understanding the complexity of metabolic mechanisms in aging and provide insight for aging and health span.
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Cuerpo Estriado/metabolismo , Hipocampo/metabolismo , Corteza Prefrontal/metabolismo , Envejecimiento/fisiología , Animales , Cromatografía Líquida de Alta Presión/métodos , Cuerpo Estriado/fisiología , Glutatión/metabolismo , Hipocampo/fisiología , Masculino , Metaboloma , Corteza Prefrontal/fisiología , Ratas Sprague-Dawley , Esfingolípidos/metabolismo , Espectrometría de Masas en Tándem/métodos , Tirosina/metabolismoRESUMEN
Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra projecting to the striatum. It has been estimated that approximately 80% of the striatal dopamine and 50% of nigral dopaminergic neurons are lost before the onset of typical motor symptoms, indicating that early diagnosis of PD using noninvasive imaging is feasible. Fluorine-19 (19 F) magnetic resonance imaging (MRI) represents a highly sensitive, easily available, low-background, and cost-effective approach to evaluate dopaminergic function using non-radioactive fluorine-containing dopaminergic agents. The aim of this study was to find a potent 19 F MRI probe to evaluate dopaminergic presynaptic function in the striatum. To select candidates for 19 F MRI probes, we investigated the following eight non-radioactive fluorine-containing dopaminergic agents: fluorodopa (F-DOPA), F-tyrosine, haloperidol, GBR13069 duhydrochloride, GBR12909 duhydrochloride, 3-bis-(4-fluorophenyl) methoxytropane hydrochloride, flupenthixol, and fenfluramine. In 19 F nuclear magnetic resonance measurements, F-tyrosine and F-DOPA displayed a relatively higher signal-to-noise ratio value in brain homogenates than in others. F-DOPA, but not F-tyrosine, induced the rotational behavior in a 6-hydroxydopamine (6-OHDA)-induced hemiparkinsonian rat model. In addition, a significantly high amount of F-DOPA accumulated in the ipsilateral striatum of hemiparkinsonian rats after the injection. We performed 19 F MRI in PC12 cells and isolated rat brain using a 7T MR scanner. Our findings suggest that F-DOPA is a promising 19 F MRI probe for evaluating dopaminergic presynaptic function in the striatum of hemiparkinsonian rats. © 2016 Wiley Periodicals, Inc.
Asunto(s)
Cuerpo Estriado/diagnóstico por imagen , Dihidroxifenilalanina/análogos & derivados , Modelos Animales de Enfermedad , Dopamina/fisiología , Imagen por Resonancia Magnética con Fluor-19/métodos , Enfermedad de Parkinson/diagnóstico por imagen , Animales , Cuerpo Estriado/metabolismo , Dihidroxifenilalanina/metabolismo , Neuronas Dopaminérgicas/fisiología , Flúor/metabolismo , Radioisótopos de Flúor/metabolismo , Masculino , Células PC12 , Enfermedad de Parkinson/metabolismo , Ratas , Ratas WistarRESUMEN
DJ-1, encoded in a causative gene of familial Parkinson's disease (PARK7), has multiple functions: it works as an antioxidant, in transcriptional regulation, as a molecular chaperone and in protein degradation. Three types of pathogenic mutants of DJ-1 (M26I, D149A and L166P) have been reported to disrupt proper structures and lead to a loss of function. DJ-1 receives oxidation at the cysteine residue, and the degree of oxidation at the C106 residue determines DJ-1 activity. In this decade, DJ-1 has been reported to suppress the progression of various neurodegenerative disorders in animal models. The administration of recombinant wild-type DJ-1 protein suppresses the neuronal loss associated with both Parkinson's disease and ischemic stroke in rats. Furthermore, in studies focused on DJ-1 as the therapeutic target, compounds that have the capacity of binding to DJ-1 at the C106 residue have been reported to exert therapeutic effects on various neurodegenerative disorders such as Parkinson's disease, Alzheimer's disease and ischemic stroke. DJ-1 and DJ-1-targeting molecules/compounds will be useful therapeutic targets for various neurodegenerative disorders due to their various functions such as antioxidant capacity, chaperone function and as a proteolytic pathway.
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Enfermedades Neurodegenerativas/tratamiento farmacológico , Enfermedades Neurodegenerativas/genética , Proteína Desglicasa DJ-1/efectos de los fármacos , Proteína Desglicasa DJ-1/genética , Animales , Antiparkinsonianos/uso terapéutico , Humanos , Enfermedad de Parkinson/tratamiento farmacológico , Enfermedad de Parkinson/genéticaRESUMEN
Parkinson's disease (PD) is a progressive neurodegenerative disorder that is primarily characterized by the degeneration of dopaminergic neurons in the nigrostriatal pathway. Loss-of-function mutations in the gene encoding PARK7/DJ-1 were identified in familial PD. Wild-type DJ-1 acts as an oxidative stress sensor in neural cells. Previously, we identified binding compounds of DJ-1, including UCP0045037/compound A, UCP0054278/compound B, and compound-23 (comp-23), by in silico virtual screening. These compounds prevented oxidative stress-induced dopaminergic neuronal death and restored locomotion defects in animal models of PD. In addition, these binding partners reduced infarct size in cerebral ischemia in rats. The neuroprotective effects of these compounds are lost in DJ-1-knockdown cells and DJ-1-knockout animal. These results suggest that these compounds interact with endogenous DJ-1 and then produce antioxidant and neuroprotective responses in both animal models for PD and cerebral ischemia in rats. This raises the possibility that interaction partners of DJ-1, such as UCP0045037, UCP0054278, and comp-23, may represent a novel dopaminergic neuroprotective drug for the treatment of PD.
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Benzamidas/farmacología , Benzodioxoles/farmacología , Enfermedades Neurodegenerativas/prevención & control , Estrés Oxidativo/efectos de los fármacos , Proteína Desglicasa DJ-1/farmacología , Animales , Benzamidas/metabolismo , Benzodioxoles/metabolismo , Isquemia Encefálica/metabolismo , Isquemia Encefálica/prevención & control , Línea Celular Tumoral , Modelos Animales de Enfermedad , Neuronas Dopaminérgicas/efectos de los fármacos , Neuronas Dopaminérgicas/metabolismo , Humanos , Enfermedades Neurodegenerativas/metabolismo , Fármacos Neuroprotectores/metabolismo , Fármacos Neuroprotectores/farmacología , Enfermedad de Parkinson/metabolismo , Enfermedad de Parkinson/prevención & control , Unión Proteica , Proteína Desglicasa DJ-1/metabolismo , Ratas , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Mitochondrial ferritin (FtMt) is a novel ferritin that sequesters iron and plays a protective role against oxidative stress. FtMt shares a high homology with H-ferritin but is expressed only in the brain, heart, and testis. In the midbrain, FtMt expression is observed in the substantia nigra. FtMt plays a neuroprotective role in the pathology of neurodegenerative diseases such as Parkinson's disease, where excessive iron induces oxidative stress, causing cell death. Herein, we investigated FtMt immunoreactivity in the brains of patients with subarachnoid hemorrhage (SAH). Double immunofluorescence labeling of tyrosine hydroxylase (TH) and FtMt showed high colocalization in the substantia nigra pars compacta (SNc) in control and SAH cases. However, in SAH cases, FtMt immunoreactivity was observed in some TH-negative neurons. Double immunofluorescence labeling of glial cell markers and FtMt showed no apparent colocalization. The number and ratio of FtMt-positive but TH-negative neurons significantly differed between the control and SAH groups. Prussian blue staining in SAH cases showed positive iron staining over a wide surface range and the substantia nigra. Thus, FtMt may be related to iron dynamics in the substantia nigra following subarachnoid hemorrhage.
RESUMEN
Mitochondrial ferritin (FtMt) is an endogenous iron-storage protein localized in the mitochondria. FtMt is mainly observed in restricted tissues, such as those in the testis, islets of Langerhans, and brain. Further, it may protect cells from oxidative stress in neurodegenerative diseases, including Alzheimer's disease and progressive supranuclear palsy. However, the role of FtMt in Parkinson's disease (PD) remains unclear. Therefore, the current study investigated the localization and expression level of FtMt in the midbrain of patients with PD and healthy controls using immunohistochemical techniques. FtMt immunoreactivity was mainly detected in dopaminergic neurons in the substantia nigra pars compacta (SNc) in both healthy controls and patients with PD. In addition, FtMt-positive particles were observed outside the dopaminergic neurons in patients with PD. Based on a quantitative comparison, patients with PD had a significantly upregulated FtMt immunoreactivity in dopaminergic neurons than healthy controls. Our result might be helpful in future studies on the role of FtMt in PD.
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In the pathogenesis of Alzheimer's disease (AD), highly neurotoxic amyloid-ß (Aß) oligomers appear early, they are thus considered to be deeply involved in the onset of Alzheimer's disease. However, Aß oligomer visualization is challenging in human tissues due to their multiple forms (e.g., low- and high-molecular-weight oligomers, including protofibrils) as well as their tendency to rapidly change forms and aggregate. In this review, we present two visualization approaches for Aß oligomers in tissues: an immunohistochemical (using the monoclonal antibody TxCo1 against toxic Aß oligomer conformers) and imaging mass spectrometry using the small chemical Shiga-Y51 that specifically binds Aß oligomers. TxCo1 immunohistochemistry revealed Aß oligomer distributions in postmortem human brains with AD. Using Shiga-Y51, imaging mass spectrometry revealed Aß oligomer distributions in the brain of a transgenic mouse model for AD. These two methods would potentially contribute to elucidating the pathological mechanisms underlying AD.
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The precision of colocalization analysis is enhanced by 3D and is potentially more accurate than 2D. Even though 3D improves the visualization of colocalization analysis, rendering a colocalization model may generate a model with numerous polygons. We developed a 3D colocalization model of FtMt/LC3 followed by simplification. Double immunofluorescence staining of FtMt and LC3 was conducted, and stacked images were acquired. We used IMARIS to render the 3D colocalization model of FtMt/LC3 and further processed it with MeshLab to decimate and generate a less complex colocalization model. We examined the available simplification algorithm using MeshLab in detail and evaluated the feasibility of each procedure in generating a model with less complexity. The quality of the simplified model was subsequently assessed. MeshLab's available shaders were scrutinized to facilitate the spatial colocalization determination. Finally, we showed that QECD was the most effective method for reducing the polygonal complexity of the colocalization model without compromising its quality. In addition, we would recommend implementing the x-ray shader, which we found useful for visualizing colocalization. As 3D was found to be more accurate in quantifying colocalization, our study provides a novel and dependable method for rendering 3D models for colocalization analysis.
Asunto(s)
Imagenología Tridimensional , Imagenología Tridimensional/métodos , Rayos X , Técnica del Anticuerpo FluorescenteRESUMEN
BACKGROUND: Tripeptide Met-Lys-Pro (MKP), a component of casein hydrolysates, has effective angiotensin-converting enzyme (ACE) inhibitory activity. Brain angiotensin II enzyme activates the NADPH oxidase complex via angiotensin II receptor type 1 (AT1) and enhances oxidative stress injury. ACE inhibitors improved cognitive function in Alzheimer's disease (AD) mouse models and previous clinical trials. Thus, although undetermined, MKP may be effective against pathological amyloid-ß (Aß) accumulation-induced cognitive impairment. OBJECTIVE: The current study aimed to investigate the potential of MKP as a pharmaceutical against AD by examining MKP's effect on cognitive function and molecular changes in the brain using double transgenic (APP/PS1) mice. METHODS: Experimental procedures were conducted in APP/PS1 mice (nâ=â38) with a C57BL/6 background. A novel object recognition test was used to evaluate recognition memory. ELISA was used to measure insoluble Aß40, Aß42, and TNF-α levels in brain tissue. Immunohistochemical analysis allowed the assessment of glial cell activation in MKP-treated APP/PS1 mice. RESULTS: The novel object recognition test revealed that MKP-treated APP/PS1 mice showed significant improvement in recognition memory. ELISA of brain tissue showed that MKP significantly reduced insoluble Aß40, Aß42, and TNF-α levels. Immunohistochemical analysis indicated the suppression of the marker for microglia and reactive astrocytes in MKP-treated APP/PS1 mice. CONCLUSION: Based on these results, we consider that MKP could ameliorate pathological Aß accumulation-induced cognitive impairment in APP/PS1 mice. Furthermore, our findings suggest that MKP potentially contributes to preventing cognitive decline in AD.