RESUMO
BACKGROUND: Vacuolar protein sorting 13 homolog A (VPS13A) disease, historically known as chorea-acanthocytosis, is a rare neurodegenerative disorder caused by biallelic mutations in VPS13A, usually resulting in reduced or absent levels of its protein product, VPS13A. VPS13A localizes to contact sites between subcellular organelles, consistent with its recently identified role in lipid transfer between membranes. Mutations are associated with neuronal loss in the striatum, most prominently in the caudate nucleus, and associated marked astrogliosis. There are no other known disease-specific cellular changes (eg, protein aggregation), but autopsy reports to date have been limited, often lacking genetic or biochemical diagnostic confirmation. OBJECTIVE: The goal of this study was to characterize neuropathological findings in the brains of seven patients with VPS13A disease (chorea-acanthocytosis). METHODS: In this study, we collected brain tissues and clinical data from seven cases of VPS13A for neuropathological analysis. The clinical diagnosis was confirmed by the presence of VPS13A mutations and/or immunoblot showing the loss or reduction of VPS13A protein. Tissues underwent routine, special, and immunohistochemical staining focused on neurodegeneration. Electron microscopy was performed in one case. RESULTS: Gross examination showed severe striatal atrophy. Microscopically, there was neuronal loss and astrogliosis in affected regions. Luxol fast blue staining showed variable lipid accumulation with diverse morphology, which was further characterized by electron microscopy. In some cases, rare degenerating p62- and ubiquitin-positive cells were present in affected regions. Calcifications were present in four cases, being extensive in one. CONCLUSIONS: We present the largest autopsy series of biochemically and genetically confirmed VPS13A disease and identify novel histopathological findings implicating abnormal lipid accumulation. © 2023 International Parkinson and Movement Disorder Society.
Assuntos
Neuroacantocitose , Humanos , Autopsia , Núcleo Caudado/metabolismo , Gliose , Lipídeos , Neuroacantocitose/genética , Neuroacantocitose/diagnóstico , Neuroacantocitose/patologia , Proteínas de Transporte Vesicular/genéticaRESUMO
In many small communities in the Mediterranean area, groundwater is usually the only water body available. Depending mainly on the surrounding geology, their concentration of naturally occurring radionuclides may pose a radiological hazard. Removal of uranium and radium from drinking water is the best way to avoid it, i.e., reverse osmosis (RO), but consuming a lot of energy. Thus, two modified drinking water treatment plants (DWTPs) using zeolites coated with manganese dioxide as adsorbent material were analyzed as an alternative to RO. Groundwater salinity can negatively affect this process. Radium removal decreased as water salinity increased; but it had a major impact on uranium, rendering the adsorption effectless in one DWTP. Waste management and how to avoid it from becoming radioactive are of major concern. Radium and uranium were associated to the reducible fraction in the filter material and also to the carbonate fraction in the case of uranium. Regeneration of the filter material using KCl solutions was able to remove 81% and 63% of uranium and radium, respectively.
Assuntos
Água Potável , Água Subterrânea , Radioatividade , Rádio (Elemento) , Urânio , Poluentes Radioativos da Água , Rádio (Elemento)/análise , Urânio/análise , Adsorção , Salinidade , Poluentes Radioativos da Água/análiseRESUMO
Developmental dyslexia is a common neurodevelopmental disorder characterized by difficulties in reading and writing. Although underlying biological and genetic mechanisms remain unclear, anomalies in phonological processing and auditory processing have been associated with dyslexia. Several candidate risk genes have also been identified, with KIAA0319 as a main candidate. Animal models targeting the rodent homolog (Kiaa0319) have been used to explore putative behavioral and anatomic anomalies, with mixed results. For example after downregulation of Kiaa0319 expression in rats via shRNA, significant adult rapid auditory processing impairments were reported, along with cortical anomalies reflecting atypical neuronal migration. Conversely, Kiaa0319 knockout (KO) mice were reported to have typical adult auditory processing, and no visible cortical anomalies. To address these inconsistencies, we tested Kiaa0319 KO mice on auditory processing tasks similar to those used previously in rat shRNA knockdown studies. Subsequent neuroanatomic analyses on these same mice targeted medial geniculate nucleus (MGN), a receptive communication-related brain structure. Results confirm that Kiaa0319 KO mice exhibit significant auditory processing impairments specific to rapid/brief stimuli, and also show significant volumetric reductions and a shift toward fewer large and smaller neurons in the MGN. The latter finding is consistent with post mortem MGN data from human dyslexic brains. Combined evidence supports a role for KIAA0319 in the development of auditory CNS pathways subserving rapid auditory processing functions critical to the development of speech processing, language, and ultimately reading. Results affirm KIAA0319 variation as a possible risk factor for dyslexia specifically via anomalies in central acoustic processing pathways.
Assuntos
Dislexia , Corpos Geniculados , Animais , Percepção Auditiva/genética , Dislexia/genética , Camundongos , Camundongos Knockout , RNA Interferente Pequeno , RatosRESUMO
The in situtechnique for measuring radionuclides in the soil using a portable Ge detector is a highly versatile tool for both the radiological characterisation and for the monitoring of operating nuclear power plants. The main disadvantage of this technique is related to the lack of knowledge of the geometry of the source whose activity concentration is to be determined. However, its greatest advantage is the high spatial representability of the samples and the reduced time and resource consumption compared to gamma spectrometry laboratory measurements. In this study, the possibilities and limits offered byin situgamma spectrometry with a high-resolution gamma portable detector in two common uses are shown. First, the radiological background characterisation and its relationship with the geology of an area of 2700 km2are assessed. Second, its potential for monitoring man-made activity concentration in soils located around an operating nuclear power plant in Spain for surveillance purposes is evaluated. Finally, high-accuracy radiation maps were prepared from the measurements that were carried out. These radiation maps are essential tools to know the radioactive background of an area, especially useful to assess artificial radioactive deposits produced after a nuclear accident or incident.
Assuntos
Monitoramento de Radiação , Poluentes Radioativos do Solo , Humanos , Centrais Nucleares , Monitoramento de Radiação/métodos , Radioisótopos/análise , Poluentes Radioativos do Solo/análise , Espectrometria gama/métodosRESUMO
The VPS13 gene family consists of VPS13A-D in mammals. Although all four genes have been linked to human diseases, their cellular functions are poorly understood, particularly those of VPS13D. We generated and characterized knockouts of each VPS13 gene in HeLa cells. Among the individual knockouts, only VPS13D-KO cells exhibit abnormal mitochondrial morphology. Additionally, VPS13D loss leads to either partial or complete peroxisome loss in several transformed cell lines and in fibroblasts derived from a VPS13D mutation-carrying patient with recessive spinocerebellar ataxia. Our data show that VPS13D regulates peroxisome biogenesis.
Assuntos
Peroxissomos/genética , Peroxissomos/metabolismo , Proteínas/genética , Proteínas/metabolismo , Células HEK293 , Células HeLa , Humanos , Mitocôndrias/genética , Mitocôndrias/metabolismo , Mutação/genéticaRESUMO
BACKGROUND: Chorea-acanthocytosis (ChAc; OMIM #200150) is a rare autosomal recessive condition with onset in early adulthood that is caused by mutations in the vacuolar protein sorting 13A (VPS13A) gene encoding chorein. Several diagnostic genomic DNA (gDNA) sequencing approaches are widely used. However, their limitations appear not to be acknowledged thoroughly enough. METHODS: Clinically, we deployed magnetic resonance imaging, blood smear analysis, and clinical chemistry for the index patient's characterization. The molecular analysis of the index patient next to his parents covered genomic DNA (gDNA) sequencing approaches, RNA/cDNA sequencing, and chorein specific Western blot. RESULTS: We report a 33-year-old male patient without functional protein due to compound heterozygosity for two VPS13A large deletions of 1168 and 1823 base pairs (bp) affecting, respectively, exons 8 and 9, and exon 13. To our knowledge, this represents the first ChAc case with two compound heterozygous large deletions identified so far. Of note, standard genomic DNA (gDNA) Sanger sequencing approaches alone yielded false negative findings. CONCLUSION: Our case demonstrates the need to carry out detection of chorein in patients suspected of having ChAc as a helpful and potentially decisive tool to establish diagnosis. Furthermore, the course of the molecular analysis in this case discloses diagnostic pitfalls in detecting some variations, such as deletions, using only standard genomic DNA (gDNA) Sanger sequencing approaches and exemplifies alternative methods, such as RNA/cDNA sequencing or qRT-PCR analysis, necessary to avoid false negative results.
Assuntos
Deleção de Genes , Testes Genéticos/métodos , Neuroacantocitose/genética , Proteínas de Transporte Vesicular/genética , Adulto , Western Blotting/métodos , Heterozigoto , Humanos , Masculino , Neuroacantocitose/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Proteínas de Transporte Vesicular/metabolismoRESUMO
The VPS13A gene is associated with the neurodegenerative disorder Chorea Acanthocytosis. It is unknown what the consequences are of impaired function of VPS13A at the subcellular level. We demonstrate that VPS13A is a peripheral membrane protein, associated with mitochondria, the endoplasmic reticulum and lipid droplets. VPS13A is localized at sites where the endoplasmic reticulum and mitochondria are in close contact. VPS13A interacts with the ER residing protein VAP-A via its FFAT domain. Interaction with mitochondria is mediated via its C-terminal domain. In VPS13A-depleted cells, ER-mitochondria contact sites are decreased, mitochondria are fragmented and mitophagy is decreased. VPS13A also localizes to lipid droplets and affects lipid droplet motility. In VPS13A-depleted mammalian cells lipid droplet numbers are increased. Our data, together with recently published data from others, indicate that VPS13A is required for establishing membrane contact sites between various organelles to enable lipid transfer required for mitochondria and lipid droplet related processes.
Assuntos
Retículo Endoplasmático/genética , Gotículas Lipídicas/metabolismo , Mitocôndrias/genética , Proteínas de Transporte Vesicular/genética , Retículo Endoplasmático/metabolismo , Endossomos/genética , Humanos , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Neuroacantocitose/genética , Doenças Neurodegenerativas/genética , Domínios Proteicos , Proteínas de Transporte Vesicular/metabolismoRESUMO
The capacity for language is one of the key features underlying the complexity of human cognition and its evolution. However, little is known about the neurobiological mechanisms that mediate normal or impaired linguistic ability. For developmental dyslexia, early postmortem studies conducted in the 1980s linked the disorder to subtle defects in the migration of neurons in the developing neocortex. These early studies were reinforced by human genetic analyses that identified dyslexia susceptibility genes and subsequent evidence of their involvement in neuronal migration. In this review, we examine recent experimental evidence that does not support the link between dyslexia and neuronal migration. We critically evaluate gene function studies conducted in rodent models and draw attention to the lack of robust evidence from histopathological and imaging studies in humans. Our review suggests that the neuronal migration hypothesis of dyslexia should be reconsidered, and the neurobiological basis of dyslexia should be approached with a fresh start.
Assuntos
Movimento Celular , Modelos Animais de Doenças , Dislexia/etiologia , Dislexia/genética , Predisposição Genética para Doença/genética , Neocórtex/citologia , Neurônios/citologia , Animais , HumanosRESUMO
Study of knockout (KO) mice has helped understand the link between many genes/proteins and human diseases. Identification of infertile KO mice provides valuable tools to characterize the molecular mechanisms underlying gamete formation. The KIAA0319L gene has been described to have a putative association with dyslexia; surprisingly, we observed that homozygous KO males for AU040320, KIAA0319L ortholog, are infertile and present a globozoospermia-like phenotype. Mutant spermatozoa are mostly immotile and display a malformed roundish head with no acrosome. In round spermatids, proacrosomal vesicles accumulate close to the acroplaxome but fail to coalesce into a single acrosomal vesicle. In wild-type mice AU040320 localises to the trans-Golgi-Network of germ cells but cannot be detected in mature acrosomes. Our results suggest AU040320 may be necessary for the normal formation of proacrosomal vesicles or the recruitment of cargo proteins required for downstream events leading to acrosomal fusion. Mutations in KIAA0319L could lead to human infertility; we screened for KIAA0319L mutations in a selected cohort of globozoospermia patients in which no genetic abnormalities have been previously identified, but detected no pathogenic changes in this particular cohort.
Assuntos
Acrossomo/metabolismo , Homozigoto , Infertilidade Masculina/genética , Proteínas de Membrana/genética , Mutação , Animais , Humanos , Masculino , Camundongos , Camundongos Knockout , Receptores de Superfície Celular/genética , Espermatogênese , Espermatozoides , Teratozoospermia/etiologia , Teratozoospermia/genéticaRESUMO
Knowledge of soil-to-plant transfer processes is a key element that can have a significant health impact. Much effort has been taken to characterize the speciation of anthropogenic radionuclides released into the environment. However, the information about naturally occurring radionuclides is scarce. This work evaluate the potential risks of transference, that is, the bioavailability of the 234,238U, 226Ra, 228,230,232Th, and 210Po in three different soils collected in Mediterranean ecosystems. Chemical speciation of these radionuclides was carried out according to two different methods, Pavlotskaya and a modification of Tessier's protocol. Most of these radionuclides were associated to fractions strongly bound to soil particles and not able to be transferred. Increasing concentrations of U and Th extracted with increasing volume of NH4OAc 1 M were observed, until it reached saturation. Readily bioavailable fraction in both methods (either exchangeable or water soluble + exchangeable) decreased in the following order: 226Ra > 234,238 U > 228,230,232Th > 210Po. It was found that < 3% of the natural radionuclide concentration in soil are readily bioavailable for plant uptake in this region of Spain, and the resulting human health risk is negligible from natural radionuclide ingestion.
Assuntos
Monitoramento Ambiental/métodos , Radioisótopos/análise , Poluentes Radioativos do Solo/análise , Disponibilidade Biológica , Humanos , Plantas/metabolismo , Radioisótopos/metabolismo , Poluentes Radioativos do Solo/metabolismo , EspanhaRESUMO
Developmental dyslexia is a neurodevelopmental disorder that affects reading ability caused by genetic and non-genetic factors. Amongst the susceptibility genes identified to date, KIAA0319 is a prime candidate. RNA-interference experiments in rats suggested its involvement in cortical migration but we could not confirm these findings in Kiaa0319-mutant mice. Given its homologous gene Kiaa0319L (AU040320) has also been proposed to play a role in neuronal migration, we interrogated whether absence of AU040320 alone or together with KIAA0319 affects migration in the developing brain. Analyses of AU040320 and double Kiaa0319;AU040320 knockouts (dKO) revealed no evidence for impaired cortical lamination, neuronal migration, neurogenesis or other anatomical abnormalities. However, dKO mice displayed an auditory deficit in a behavioral gap-in-noise detection task. In addition, recordings of click-evoked auditory brainstem responses revealed suprathreshold deficits in wave III amplitude in AU040320-KO mice, and more general deficits in dKOs. These findings suggest that absence of AU040320 disrupts firing and/or synchrony of activity in the auditory brainstem, while loss of both proteins might affect both peripheral and central auditory function. Overall, these results stand against the proposed role of KIAA0319 and AU040320 in neuronal migration and outline their relationship with deficits in the auditory system.
Assuntos
Percepção Auditiva/fisiologia , Movimento Celular/fisiologia , Córtex Cerebral/metabolismo , Proteínas do Tecido Nervoso/deficiência , Neurônios/metabolismo , Receptores de Superfície Celular/deficiência , Potenciais de Ação/fisiologia , Adaptação Fisiológica/fisiologia , Animais , Córtex Cerebral/crescimento & desenvolvimento , Córtex Cerebral/patologia , Dislexia/genética , Potenciais Evocados Auditivos do Tronco Encefálico/fisiologia , Feminino , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas do Tecido Nervoso/genética , Neurogênese/fisiologia , Neurônios/patologia , Receptores de Superfície Celular/genéticaRESUMO
KIAA0319 is a transmembrane protein associated with dyslexia with a presumed role in neuronal migration. Here we show that KIAA0319 expression is not restricted to the brain but also occurs in sensory and spinal cord neurons, increasing from early postnatal stages to adulthood and being downregulated by injury. This suggested that KIAA0319 participates in functions unrelated to neuronal migration. Supporting this hypothesis, overexpression of KIAA0319 repressed axon growth in hippocampal and dorsal root ganglia neurons; the intracellular domain of KIAA0319 was sufficient to elicit this effect. A similar inhibitory effect was observed in vivo as axon regeneration was impaired after transduction of sensory neurons with KIAA0319. Conversely, the deletion of Kiaa0319 in neurons increased neurite outgrowth in vitro and improved axon regeneration in vivo. At the mechanistic level, KIAA0319 engaged the JAK2-SH2B1 pathway to activate Smad2, which played a central role in KIAA0319-mediated repression of axon growth. In summary, we establish KIAA0319 as a novel player in axon growth and regeneration with the ability to repress the intrinsic growth potential of axons. This study describes a novel regulatory mechanism operating during peripheral nervous system and central nervous system axon growth, and offers novel targets for the development of effective therapies to promote axon regeneration.
Assuntos
Axônios/metabolismo , Moléculas de Adesão Celular/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Crescimento Neuronal , Proteína Smad2/metabolismo , Envelhecimento/metabolismo , Animais , Crescimento Celular , Linhagem Celular , Células Cultivadas , Feminino , Gânglios Espinais/metabolismo , Hipocampo/metabolismo , Humanos , Janus Quinase 2/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Regeneração Nervosa/fisiologia , Proteínas do Tecido Nervoso/genética , Neurônios/metabolismo , Domínios Proteicos , Ratos Wistar , Nervo Isquiático/lesões , Nervo Isquiático/metabolismo , Medula Espinal/metabolismoRESUMO
Chorea-Acanthocytosis (ChAc) is a rare hereditary neurological disorder characterized by abnormal movements, red blood cell pathology, and progressive neurodegeneration. Little is understood of the pathogenesis of ChAc and related disorders (collectively Neuroacanthocytosis). The Eighth International Chorea-Acanthocytosis Symposium was held in May 2016 in Ann Arbor, MI, USA, and focused on molecular mechanisms driving ChAc pathophysiology. Accompanying the meeting, members of the neuroacanthocytosis research community and other invited scientists met in a workshop to discuss the current understanding and next steps needed to better understand ChAc pathogenesis. These discussions identified several broad and critical needs for advancing ChAc research and patient care, and led to the definition of 18 specific action points related to functional and molecular studies, animal models, and clinical research. These action points, described below, represent tractable research goals to pursue for the next several years.
RESUMO
Chorea-Acanthocytosis is a rare, neurodegenerative disorder characterized by progressive loss of locomotor and cognitive function. It is caused by loss of function mutations in the Vacuolar Protein Sorting 13A (VPS13A) gene, which is conserved from yeast to human. The consequences of VPS13A dysfunction in the nervous system are still largely unspecified. In order to study the consequences of VPS13A protein dysfunction in the ageing central nervous system we characterized a Drosophila melanogaster Vps13 mutant line. The Drosophila Vps13 gene encoded a protein of similar size as human VPS13A. Our data suggest that Vps13 is a peripheral membrane protein located to endosomal membranes and enriched in the fly head. Vps13 mutant flies showed a shortened life span and age associated neurodegeneration. Vps13 mutant flies were sensitive to proteotoxic stress and accumulated ubiquitylated proteins. Levels of Ref(2)P, the Drosophila orthologue of p62, were increased and protein aggregates accumulated in the central nervous system. Overexpression of the human Vps13A protein in the mutant flies partly rescued apparent phenotypes. This suggests a functional conservation of human VPS13A and Drosophila Vps13. Our results demonstrate that Vps13 is essential to maintain protein homeostasis in the larval and adult Drosophila brain. Drosophila Vps13 mutants are suitable to investigate the function of Vps13 in the brain, to identify genetic enhancers and suppressors and to screen for potential therapeutic targets for Chorea-Acanthocytosis.
Assuntos
Encéfalo/fisiologia , Proteínas de Drosophila/fisiologia , Homeostase/fisiologia , Proteínas do Tecido Nervoso/fisiologia , Proteínas de Transporte Vesicular/fisiologia , Animais , Encéfalo/patologia , Drosophila , Proteínas de Drosophila/genética , Humanos , Mutação , Proteínas de Transporte Vesicular/genéticaRESUMO
Developmental dyslexia is a common disorder with a strong genetic component, but the underlying molecular mechanisms are still unknown. Several candidate dyslexia-susceptibility genes, including KIAA0319, DYX1C1, and DCDC2, have been identified in humans. RNA interference experiments targeting these genes in rat embryos have shown impairments in neuronal migration, suggesting that defects in radial cortical migration could be involved in the disease mechanism of dyslexia. Here we present the first characterisation of a Kiaa0319 knockout mouse line. Animals lacking KIAA0319 protein do not show anatomical abnormalities in any of the layered structures of the brain. Neurogenesis and radial migration of cortical projection neurons are not altered, and the intrinsic electrophysiological properties of Kiaa0319-deficient neurons do not differ from those of wild-type neurons. Kiaa0319 overexpression in cortex delays radial migration, but does not affect final neuronal position. However, knockout animals show subtle differences suggesting possible alterations in anxiety-related behaviour and in sensorimotor gating. Our results do not reveal a migration disorder in the mouse model, adding to the body of evidence available for Dcdc2 and Dyx1c1 that, unlike in the rat in utero knockdown models, the dyslexia-susceptibility candidate mouse homolog genes do not play an evident role in neuronal migration. However, KIAA0319 protein expression seems to be restricted to the brain, not only in early developmental stages but also in adult mice, indicative of a role of this protein in brain function. The constitutive and conditional knockout lines reported here will be useful tools for further functional analyses of Kiaa0319.
Assuntos
Movimento Celular/genética , Dislexia/genética , Dislexia/patologia , Neocórtex/patologia , Proteínas do Tecido Nervoso/deficiência , Neurônios/fisiologia , Fatores Etários , Animais , Animais Recém-Nascidos , Ansiedade/etiologia , Ansiedade/genética , Encéfalo/metabolismo , Adaptação à Escuridão/genética , Modelos Animais de Doenças , Dislexia/complicações , Eletroporação , Embrião de Mamíferos , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Genótipo , Técnicas In Vitro , Antígeno Ki-67/metabolismo , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Masculino , Potenciais da Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neocórtex/metabolismo , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Neurogênese/genética , Fator de Transcrição PAX6/metabolismo , Técnicas de Patch-Clamp , Gravidez , Inibição Pré-Pulso/genética , Interferência de RNA , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Filtro Sensorial/genética , Proteínas com Domínio T/metabolismo , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismoRESUMO
The anthropogenic (137Cs, 90Sr, 239+240Pu and 241Am) and naturally occurring radionuclide (40K, 234,238U, 228,230,232Th, 226Ra and 210Pb) content in near surface air present seasonal variations related to natural processes, such as soil erosion, resuspension of fine particles of soil and radon exhalation from soil (210Pb). The objective is to analyze seasonal variations of their concentrations and compare with radiological events (Fukushima fallout and wild fire) in a location without any known source of anthropogenic radionuclides. The 210Pb, 40K, and 137Cs presented annual variations, with maximum activity levels in summer. Solar radiation and rainfall were correlated with 210Pb and 40K. The 234,238U, 228,230,232Th, 226Ra, 137Cs and 90Sr presented positive correlation with monthly mean values of temperature. The ratio 90Sr/137Cs was within the range of those reported for soils in Spain. Finally, the maximal effective dose rate was estimated to be 37 and 88 µSv/y for infants and adults, respectively, well below 1 mSv/y reference level. The main contributor to effective dose was 210Pb, about 92%, followed by: 210Pb â« 228,230,232Th > 226Ra, 234,238U > 7Be, 239+240Pu > 40K, 90Sr > 137Cs > 22Na.
Assuntos
Poluentes Radioativos do Ar/análise , Contaminação Radioativa do Ar/estatística & dados numéricos , Exposição à Radiação/estatística & dados numéricos , Monitoramento de Radiação , Radioisótopos/análise , Adulto , Humanos , Lactente , Recém-Nascido , Espanha , Inquéritos e QuestionáriosRESUMO
Single nucleotide polymorphisms (SNPs) close to the VPS13C, C2CD4A and C2CD4B genes on chromosome 15q are associated with impaired fasting glucose and increased risk of type 2 diabetes. eQTL analysis revealed an association between possession of risk (C) alleles at a previously implicated causal SNP, rs7163757, and lowered VPS13C and C2CD4A levels in islets from female (n = 40, P < 0.041) but not from male subjects. Explored using promoter-reporter assays in ß-cells and other cell lines, the risk variant at rs7163757 lowered enhancer activity. Mice deleted for Vps13c selectively in the ß-cell were generated by crossing animals bearing a floxed allele at exon 1 to mice expressing Cre recombinase under Ins1 promoter control (Ins1Cre). Whereas Vps13c(fl/fl):Ins1Cre (ßVps13cKO) mice displayed normal weight gain compared with control littermates, deletion of Vps13c had little effect on glucose tolerance. Pancreatic histology revealed no significant change in ß-cell mass in KO mice vs. controls, and glucose-stimulated insulin secretion from isolated islets was not altered in vitro between control and ßVps13cKO mice. However, a tendency was observed in female null mice for lower insulin levels and ß-cell function (HOMA-B) in vivo. Furthermore, glucose-stimulated increases in intracellular free Ca(2+) were significantly increased in islets from female KO mice, suggesting impaired Ca(2+) sensitivity of the secretory machinery. The present data thus provide evidence for a limited role for changes in VPS13C expression in conferring altered disease risk at this locus, particularly in females, and suggest that C2CD4A may also be involved.
Assuntos
Proteínas de Ligação ao Cálcio/genética , Intolerância à Glucose/genética , Células Secretoras de Insulina/metabolismo , Proteínas do Tecido Nervoso/genética , Proteínas/genética , Animais , Western Blotting , Cálcio/metabolismo , Diabetes Mellitus Tipo 2/genética , Feminino , Células Secretoras de Glucagon/patologia , Insulina/metabolismo , Resistência à Insulina , Secreção de Insulina , Células Secretoras de Insulina/patologia , Masculino , Camundongos , Camundongos Knockout , Pâncreas/patologia , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase em Tempo Real , Fatores Sexuais , Proteínas de Transporte VesicularRESUMO
Galectin-12, a member of the galectin family of ß-galactoside-binding animal lectins, is preferentially expressed in adipocytes and required for adipocyte differentiation in vitro. This protein was recently found to regulate lipolysis, whole body adiposity, and glucose homeostasis in vivo. Here we identify VPS13C, a member of the VPS13 family of vacuolar protein sorting-associated proteins highly conserved throughout eukaryotic evolution, as a major galectin-12-binding protein. VPS13C is upregulated during adipocyte differentiation, and is required for galectin-12 protein stability. Knockdown of Vps13c markedly reduces the steady-state levels of galectin-12 by promoting its degradation through primarily the lysosomal pathway, and impairs adipocyte differentiation. Our studies also suggest that VPS13C may have a broader role in protein quality control. The regulation of galectin-12 stability by VPS13C could potentially be exploited for therapeutic intervention of obesity and related metabolic diseases.
Assuntos
Adipogenia/genética , Galectinas/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Proteínas/metabolismo , Células 3T3-L1 , Animais , Galectinas/genética , Camundongos , Proteínas do Tecido Nervoso/genética , Estabilidade Proteica , Proteínas/genética , Proteínas de Transporte VesicularRESUMO
Naturally Occurring Radioactive Materials (NORM) wastes are generated in huge quantities in several industries and their management has been carried out under considerations of industrial non-radioactive wastes, before the concern on the radioactivity content was included in the legislation. Therefore these wastes were conditioned using conventional methods and the waste disposals were designed to isolate toxic elements from the environment for long periods of time. Spanish regulation for these conventional toxic waste disposals includes conditions that assure adequate isolation to minimize the impact of the wastes to the environment in present and future conditions. After 1996 the radiological impact of the management of NORM wastes is considered and all the aspects related with natural radiations and the radiological control regarding the management of residues from NORM industries were developed in the new regulation. One option to be assessed is the disposal of NORM wastes in hazardous and non-hazardous waste disposals, as was done before this new regulation. This work analyses the management of NORM wastes in these landfills to derive the masses that can be disposed without considerable radiological impact. Generic dose assessments were carried out under highly conservative hypothesis and a discussion on the uncertainty and variability sources was included to provide consistency to the calculations.