RESUMO
The neuronal ceroid lipofuscinoses (NCLs) are at least 13 distinct progressive neurodegenerative disorders unified by the accumulation of lysosomal auto-fluorescent material called lipofuscin. The only form that occurs via autosomal-dominant inheritance exhibits adult onset and is sometimes referred to as Parry type NCL. The manifestations may include behavioral symptoms followed by seizures, ataxia, dementia, and early death. Mutations in the gene DNAJC5 that codes for the presynaptic co-chaperone cysteine string protein-α (CSPα) were recently reported in sporadic adult-onset cases and in families with dominant inheritance. The mutant CSPα protein may lead to disease progression by both loss and gain of function mechanisms. Iron chelation therapy may be considered as a possible pharmaceutical intervention based on our recent mechanism-based proposal of CSPα oligomerization via ectopic Fe-S cluster-binding, summarized in this review.
Assuntos
Proteínas de Choque Térmico HSP40/genética , Proteínas de Membrana/genética , Lipofuscinoses Ceroides Neuronais/genética , Lipofuscinoses Ceroides Neuronais/terapia , Neurônios/metabolismo , Genes Dominantes/genética , Humanos , Ferro/metabolismo , Quelantes de Ferro/uso terapêutico , Mutação/genética , Lipofuscinoses Ceroides Neuronais/fisiopatologia , Neurônios/patologia , LinhagemRESUMO
AIMS: The most common autosomal recessive limb girdle muscular dystrophy is associated with the CAPN3 gene. The exclusively recessive inheritance of this disorder has been recently challenged by the description of the recurrent variants, c.643_663del21 [p.(Ser215_Gly221del)] and c.598_612del15 [p.(Phe200_Leu204del)], associated with autosomal dominant inheritance. Our objective was to confirm the existence of autosomal dominant calpainopathies. METHODS: Through our activity as one of the reference centres for genetic diagnosis of calpainopathies in France and the resulting collaborations through the French National Network for Rare Neuromuscular Diseases (FILNEMUS), we identified four families harbouring the same CAPN3 heterozygous variant with supposedly autosomal dominant inheritance. RESULTS: We identified a novel dominantly inherited CAPN3 variant, c.1333G>A [p.(Gly445Arg)] in 14 affected patients from four unrelated families. The complementary phenotypic, functional and genetic findings correlate with an autosomal dominant inheritance in these families, emphasizing the existence of this novel transmission mode for calpainopathies. The mild phenotype associated with these autosomal dominant cases widens the phenotypic spectrum of calpainopathies and should therefore be considered in clinical practice. CONCLUSIONS: We confirm the existence of autosomal dominant calpainopathies as an entity beyond the cases related to the in-frame deletions c.643_663del21 and c.598_612del15, with the identification of a novel dominantly inherited and well-documented CAPN3 missense variant, c.1333G>A [p.(Gly445Arg)]. In addition to the consequences for genetic counselling, the confirmation of an autosomal dominant transmission mode for calpainopathies underlines the importance of re-assessing other myopathies for which the inheritance is considered as strictly autosomal recessive.
Assuntos
Calpaína/genética , Aberrações Cromossômicas , Proteínas Musculares/genética , Doenças Neuromusculares/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , França , Genes Dominantes/genética , Variação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Mutação/genética , Linhagem , Fenótipo , Adulto JovemRESUMO
Although genetic factors contribute to almost half of all cases of deafness, treatment options for genetic deafness are limited. We developed a genome-editing approach to target a dominantly inherited form of genetic deafness. Here we show that cationic lipid-mediated in vivo delivery of Cas9-guide RNA complexes can ameliorate hearing loss in a mouse model of human genetic deafness. We designed and validated, both in vitro and in primary fibroblasts, genome editing agents that preferentially disrupt the dominant deafness-associated allele in the Tmc1 (transmembrane channel-like gene family 1) Beethoven (Bth) mouse model, even though the mutant Tmc1Bth allele differs from the wild-type allele at only a single base pair. Injection of Cas9-guide RNA-lipid complexes targeting the Tmc1Bth allele into the cochlea of neonatal Tmc1Bth/+ mice substantially reduced progressive hearing loss. We observed higher hair cell survival rates and lower auditory brainstem response thresholds in injected ears than in uninjected ears or ears injected with control complexes that targeted an unrelated gene. Enhanced acoustic startle responses were observed among injected compared to uninjected Tmc1Bth/+ mice. These findings suggest that protein-RNA complex delivery of target gene-disrupting agents in vivo is a potential strategy for the treatment of some types of autosomal-dominant hearing loss.
Assuntos
Proteínas Associadas a CRISPR/administração & dosagem , Edição de Genes/métodos , Genes Dominantes/genética , Terapia Genética/métodos , Perda Auditiva/genética , Estimulação Acústica , Alelos , Animais , Animais Recém-Nascidos , Limiar Auditivo , Sequência de Bases , Proteínas Associadas a CRISPR/metabolismo , Proteínas Associadas a CRISPR/uso terapêutico , Sistemas CRISPR-Cas , Sobrevivência Celular , Cóclea/citologia , Cóclea/metabolismo , Modelos Animais de Doenças , Potenciais Evocados Auditivos do Tronco Encefálico , Feminino , Fibroblastos , Células Ciliadas Auditivas/citologia , Perda Auditiva/fisiopatologia , Perda Auditiva/prevenção & controle , Humanos , Lipossomos , Masculino , Proteínas de Membrana/genética , Camundongos , Reflexo de SobressaltoRESUMO
N-methyl-D-aspartate receptors (NMDARs) are ligand-gated cation channels that mediate excitatory synaptic transmission. Genetic mutations in multiple NMDAR subunits cause various childhood epilepsy syndromes. Here, we report a de novo recurrent heterozygous missense mutation-c.1999G>A (p.Val667Ile)-in a NMDAR gene previously unrecognized to harbor disease-causing mutations, GRIN2D, identified by exome and candidate panel sequencing in two unrelated children with epileptic encephalopathy. The resulting GluN2D p.Val667Ile exchange occurs in the M3 transmembrane domain involved in channel gating. This gain-of-function mutation increases glutamate and glycine potency by 2-fold, increases channel open probability by 6-fold, and reduces receptor sensitivity to endogenous negative modulators such as extracellular protons. Moreover, this mutation prolongs the deactivation time course after glutamate removal, which controls the synaptic time course. Transfection of cultured neurons with human GRIN2D cDNA harboring c.1999G>A leads to dendritic swelling and neuronal cell death, suggestive of excitotoxicity mediated by NMDAR over-activation. Because both individuals' seizures had proven refractory to conventional antiepileptic medications, the sensitivity of mutant NMDARs to FDA-approved NMDAR antagonists was evaluated. Based on these results, oral memantine was administered to both children, with resulting mild to moderate improvement in seizure burden and development. The older proband subsequently developed refractory status epilepticus, with dramatic electroclinical improvement upon treatment with ketamine and magnesium. Overall, these results suggest that NMDAR antagonists can be useful as adjuvant epilepsy therapy in individuals with GRIN2D gain-of-function mutations. This work further demonstrates the value of functionally evaluating a mutation, enabling mechanistic understanding and therapeutic modeling to realize precision medicine for epilepsy.
Assuntos
Genes Dominantes/genética , Mutação , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Espasmos Infantis/tratamento farmacológico , Espasmos Infantis/genética , Sequência de Aminoácidos , Sequência de Bases , Morte Celular , Criança , Análise Mutacional de DNA , Dendritos/patologia , Eletroencefalografia , Exoma/genética , Feminino , Ácido Glutâmico/metabolismo , Humanos , Lactente , Recém-Nascido , Ketamina/uso terapêutico , Magnésio/uso terapêutico , Memantina/administração & dosagem , Memantina/uso terapêutico , Modelos Moleculares , Medicina de Precisão , Receptores de N-Metil-D-Aspartato/química , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Convulsões/tratamento farmacológico , Convulsões/genética , Convulsões/metabolismo , Espasmos Infantis/metabolismoRESUMO
Stalk rots are destructive diseases in maize around the world, and are most often caused by the pathogen Pythium, Fusarium and other fungi. The most efficient management for controlling stalk rots is to breed resistant cultivars. Pythium stalk rot can cause serious yield loss on maize, and to find the resistance genes from the existing germplasm is the basis to develop Pythium-resistance hybrid lines. In this study, we investigated the genetic resistance to Pythium stalk rot in inbred line Qi319 using F2 and F2:3 population, and found that the resistance to Pythium inflatum in Qi319 was conferred by two independently inherited dominant genes, RpiQI319-1 and RpiQI319-2. Linkage analysis uncovered that the RpiQI319-1 co-segregated with markers bnlg1203, and bnlg2057 on chromosome 1, and that the RpiQI319-2 locus co-segregated with markers umc2069 and bnlg1716 on chromosome 10. The RpiQI319-1 locus was further mapped into a ~500-kb interval flanked by markers SSRZ33 and SSRZ47. These results will facilitate marker-assisted selection of Pythium stalk rot-resistant cultivars in maize breeding. To our knowledge, this is the first report on the resistance to P. inflatum in the inbred line Qi319, and is also the first description of two independently inherited dominant genes conferring the resistance of Pythium stalk rot in maize.
Assuntos
Resistência à Doença/genética , Genes de Plantas/genética , Doenças das Plantas/genética , Zea mays/genética , Mapeamento Cromossômico , Cromossomos de Plantas/genética , DNA de Plantas/genética , Genes Dominantes/genética , Ligação Genética , Marcadores Genéticos/genética , Interações Hospedeiro-Patógeno , Endogamia , Doenças das Plantas/microbiologia , Reação em Cadeia da Polimerase , Pythium/fisiologia , Zea mays/microbiologiaRESUMO
Genome-wide association studies (GWAS) have reproducibly associated variants within introns of FTO with increased risk for obesity and type 2 diabetes (T2D). Although the molecular mechanisms linking these noncoding variants with obesity are not immediately obvious, subsequent studies in mice demonstrated that FTO expression levels influence body mass and composition phenotypes. However, no direct connection between the obesity-associated variants and FTO expression or function has been made. Here we show that the obesity-associated noncoding sequences within FTO are functionally connected, at megabase distances, with the homeobox gene IRX3. The obesity-associated FTO region directly interacts with the promoters of IRX3 as well as FTO in the human, mouse and zebrafish genomes. Furthermore, long-range enhancers within this region recapitulate aspects of IRX3 expression, suggesting that the obesity-associated interval belongs to the regulatory landscape of IRX3. Consistent with this, obesity-associated single nucleotide polymorphisms are associated with expression of IRX3, but not FTO, in human brains. A direct link between IRX3 expression and regulation of body mass and composition is demonstrated by a reduction in body weight of 25 to 30% in Irx3-deficient mice, primarily through the loss of fat mass and increase in basal metabolic rate with browning of white adipose tissue. Finally, hypothalamic expression of a dominant-negative form of Irx3 reproduces the metabolic phenotypes of Irx3-deficient mice. Our data suggest that IRX3 is a functional long-range target of obesity-associated variants within FTO and represents a novel determinant of body mass and composition.
Assuntos
Proteínas de Homeodomínio/genética , Íntrons/genética , Oxigenases de Função Mista/genética , Obesidade/genética , Oxo-Ácido-Liases/genética , Proteínas/genética , Fatores de Transcrição/genética , Tecido Adiposo/metabolismo , Dioxigenase FTO Dependente de alfa-Cetoglutarato , Animais , Metabolismo Basal/genética , Índice de Massa Corporal , Peso Corporal/genética , Encéfalo/metabolismo , Diabetes Mellitus Tipo 2/genética , Dieta , Genes Dominantes/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Hipotálamo/metabolismo , Masculino , Camundongos , Fenótipo , Polimorfismo de Nucleotídeo Único/genética , Regiões Promotoras Genéticas/genética , Magreza/genética , Fatores de Transcrição/deficiência , Fatores de Transcrição/metabolismo , Peixe-Zebra/embriologia , Peixe-Zebra/genéticaRESUMO
UNLABELLED: Acute necrotizing encephalopathy is a rare neurologic disease most often triggered by a febrile viral event affecting an otherwise healthy infant. The clinical course is characterized by rapid deterioration of the neurological condition that often leads to coma and requires intensive care. The diagnosis is usually suggested by MRI, which shows symmetrical and focal necrotic lesions of thalami. Acute necrotizing encephalopathy has been linked in recent studies to an autosomal-dominant mutation of the gene for the protein RAN-binding protein 2. CASE REPORT: We report three cases in siblings of Tunisian origin. Two of them presented with acute necrotizing encephalopathy at the age of 9 months in the immediate aftermath of a viral infection. The molecular study conducted in the family showed that both patients and their mother were carriers of the missense mutation gene RAN-binding protein 2. COMMENTS: Although the role of Ran BP2 protein is incompletely known, mutation of the RANBP2 gene causes rare, reversible central neurologic disorders. Suspected diagnosis is facilitated by MRI, which shows specific lesions of multifocal, symmetric involvement of the thalami, brainstem tegmentum, supratentorial white matter, and cerebellum. Due to the low frequency of the disease and its non-specific clinical presentation, the diagnosis of acute necrotizing encephalopathy is a major challenge, while preventative measures can be proposed in familial mutation.
Assuntos
Análise Mutacional de DNA , Emigrantes e Imigrantes , Genes Dominantes/genética , Leucoencefalite Hemorrágica Aguda/genética , Chaperonas Moleculares/genética , Mutação de Sentido Incorreto/genética , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Cerebelo/patologia , Aberrações Cromossômicas , Diagnóstico Diferencial , Progressão da Doença , Dominância Cerebral/fisiologia , França , Triagem de Portadores Genéticos , Humanos , Lactente , Leucoencefalite Hemorrágica Aguda/diagnóstico , Imageamento por Ressonância Magnética , Masculino , Exame Neurológico , Tegmento Mesencefálico/patologia , Tálamo/patologia , Tunísia/etnologia , Viroses/complicaçõesRESUMO
Arabidopsis Fused kinase TWO-IN-ONE (TIO) controls phragmoplast expansion through its interaction with the Kinesin-12 subfamily proteins that anchor the plus ends of interdigitating microtubules in the phragmoplast midzone. Previous analyses of loss-of-function mutants and RNA interference lines revealed that TIO positively controls both somatic and gametophytic cell cytokinesis; however, knowledge of the full spectrum of TIO functions during plant development remains incomplete. To characterize TIO functions further, we expressed TIO and a range of TIO variants under control of the TIO promoter in wild-type Arabidopsis plants. We discovered that TIO-overexpressing transgenic lines produce enlarged pollen grains, arising from incomplete cytokinesis during male meiosis, and show sporophytic abnormalities indicative of polyploidy. These phenotypes arose independently in TIO variants in which either gametophytic function or the ability of TIO to interact with Kinesin-12 subfamily proteins was abolished. Interaction assays in yeast showed TIO to bind to the AtNACK2/TETRASPORE, and plants doubly homozygous for kinesin-12a and kinesin-12b knockout mutations to produce enlarged pollen grains. Our results show TIO to dominantly inhibit male meiotic cytokinesis in a dosage-dependent manner that may involve direct binding to a component of the canonical NACK-PQR cytokinesis signaling pathway.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/enzimologia , Citocinese/genética , Regulação da Expressão Gênica de Plantas , Arabidopsis/citologia , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Arabidopsis/metabolismo , Genes Dominantes/genética , Cinesinas/genética , Cinesinas/metabolismo , Meiose/genética , Microtúbulos/genética , Microtúbulos/metabolismo , Mutagênese Insercional , Fenótipo , Fosfotransferases/genética , Fosfotransferases/metabolismo , Pólen/citologia , Pólen/enzimologia , Pólen/genética , Pólen/fisiologia , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Transdução de Sinais , Técnicas do Sistema de Duplo-HíbridoRESUMO
PURPOSE: In relatively small series, autosomal dominant lateral temporal epilepsy (ADLTE) has been associated with leucine-rich, glioma-inactivated 1 (LGI1) mutations in about 50% of the families, this genetic heterogeneity being probably caused by differences in the clinical characteristics of the families. In this article we report the overall clinical and genetic spectrum of ADLTE in Italy with the aim to provide new insight into its nosology and genetic basis. METHODS: In a collaborative study of the Commission of Genetics of the Italian League Against Epilepsy (LICE) encompassing a 10-year period (2000-2010), we collected 33 ADLTE families, selected on the basis of the following criteria: presence of at least two members concordant for unprovoked partial seizures with prominent auditory and or aphasic symptoms, absence of any known structural brain pathology or etiology, and normal neurologic examination. The clinical, neurophysiologic, and neuroradiologic findings of all patients were analyzed and a genealogic tree was built for each pedigree. The probands' DNA was tested for LGI1 mutations by direct sequencing and, if negative, were genotyped with single-nucleotide polymorphism (SNP) array to search for disease-linked copy-number variation CNV. The disease penetrance in mutated and nonmutated families was assessed as a proportion of obligate carriers who were affected. KEY FINDINGS: The 33 families included a total of 127 affected individuals (61 male, 66 female, 22 deceased). The age at onset ranged between 2 and 60 years (mean 18.7 years). Ninety-one patients (72%) had clear-cut focal (elementary, complex, or secondarily generalized) seizures, characterized by prominent auditory auras in 68% of the cases. Other symptoms included complex visual hallucinations, vertigo, and déjà vu. Aphasic seizures, associated or not with auditory features, were observed in 20% of the cases, whereas tonic-clonic seizures occurred in 86% of the overall series. Sudden noises could precipitate the seizures in about 20% of cases. Seizures, which usually occurred at a low frequency, were promptly controlled or markedly improved by antiepileptic treatment in the majority of patients. The interictal electroencephalography (EEG) studies showed the epileptiform temporal abnormalities in 62% of cases, with a slight predominance over the left region. Magnetic resonance imaging (MRI) or computerized tomography (CT) scans were negative. LGI1 mutations (missense in nine and a microdeletion in one) were found in only 10 families (30%). The patients belonging to the mutated and not mutated groups did not differ except for penetrance estimate, which was 61.3% and 35% in the two groups, respectively (chi-square, p = 0.017). In addition, the disease risk of members of families with mutations in LGI1 was three times higher than that of members of LGI1-negative families (odds ratio [OR] 2.94, confidence interval [CI] 1.2-7.21). SIGNIFICANCE: A large number of ADLTE families has been collected over a 10-year period in Italy, showing a typical and homogeneous phenotype. LGI1 mutations have been found in only one third of families, clinically indistinguishable from nonmutated pedigrees. The estimate of penetrance and OR, however, demonstrates a significantly lower penetrance rate and relative disease risk in non-LGI1-mutated families compared with LGI1-mutated pedigrees, suggesting that a complex inheritance pattern may underlie a proportion of these families.
Assuntos
Epilepsia do Lobo Temporal/genética , Saúde da Família , Genes Dominantes/genética , Mutação/genética , Penetrância , Proteínas/genética , Estimulação Acústica , Adolescente , Adulto , Idade de Início , Criança , Pré-Escolar , Análise Mutacional de DNA , Eletroencefalografia , Epilepsia do Lobo Temporal/fisiopatologia , Feminino , Genótipo , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Itália , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Fenótipo , Estudos Retrospectivos , Adulto JovemRESUMO
Nocturnal frontal lobe epilepsy has been historically considered a channelopathy caused by mutations in subunits of the neuronal nicotinic acetylcholine receptor or in a recently reported potassium channel. However, these mutations account for only a minority of patients, and the existence of at least a new locus for the disease has been demonstrated. In 2005, we detected two nucleotide variations in the promoter of the CRH gene coding for the corticotropin releasing hormone in 7 patients. These variations cosegregated with the disease and were demonstrated to alter the cellular levels of this hormone. Here, we report the identification in an Italian affected family of a novel missense mutation (hpreproCRH p.Pro30Arg) located in the region of the CRH coding for the protein pro-sequence. The mutation was detected in heterozygosity in the two affected individuals. In vitro assays demonstrated that this mutation results in reduced levels of protein secretion in the short time thus suggesting that mutated people could present an altered capability to respond immediately to stress agents.
Assuntos
Hormônio Liberador da Corticotropina/genética , Epilepsia do Lobo Frontal/genética , Genes Dominantes/genética , Mutação de Sentido Incorreto/genética , Análise de Variância , Sequência de Bases , Western Blotting , Hormônio Liberador da Corticotropina/metabolismo , Primers do DNA/genética , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Epilepsia do Lobo Frontal/tratamento farmacológico , Vetores Genéticos/genética , Heterozigoto , Humanos , Itália , Levetiracetam , Imageamento por Ressonância Magnética , Microscopia de Fluorescência , Dados de Sequência Molecular , Linhagem , Piracetam/análogos & derivados , Piracetam/uso terapêutico , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNARESUMO
This study describes a dominant low-seed-oil mutant (lo15571) of Arabidopsis (Arabidopsis thaliana) generated by enhancer tagging. Compositional analysis of developing siliques and mature seeds indicated reduced conversion of photoassimilates to oil. Immunoblot analysis revealed increased levels of At1g01050 protein in developing siliques of lo15571. At1g01050 encodes a soluble, cytosolic pyrophosphatase and is one of five closely related genes that share predicted cytosolic localization and at least 70% amino acid sequence identity. Expression of At1g01050 using a seed-preferred promoter recreated most features of the lo15571 seed phenotype, including low seed oil content and increased levels of transient starch and soluble sugars in developing siliques. Seed-preferred RNA interference-mediated silencing of At1g01050 and At3g53620, a second cytosolic pyrophosphatase gene that shows expression during seed filling, led to a heritable oil increase of 1% to 4%, mostly at the expense of seed storage protein. These results are consistent with a scenario in which the rate of mobilization of sucrose, for precursor supply of seed storage lipid biosynthesis by cytosolic glycolysis, is strongly influenced by the expression of endogenous pyrophosphatase enzymes. This emphasizes the central role of pyrophosphate-dependent reactions supporting cytosolic glycolysis during seed maturation when ATP supply is low, presumably due to hypoxic conditions. This route is the major route providing precursors for seed oil biosynthesis. ATP-dependent reactions at the entry point of glycolysis in the cytosol or plastid cannot fully compensate for the loss of oil content observed in transgenic events with increased expression of cytosolic pyrophosphatase enzyme in the cytosol. These findings shed new light on the dynamic properties of cytosolic pyrophosphate pools in developing seed and their influence on carbon partitioning during seed filling. Finally, our work uniquely demonstrates that genes encoding cytosolic pyrophosphatase enzymes provide novel targets to improve seed composition for plant biotechnology applications.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Arabidopsis/crescimento & desenvolvimento , Citosol/enzimologia , Óleos de Plantas/metabolismo , Pirofosfatases/metabolismo , Sementes/crescimento & desenvolvimento , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Centrifugação com Gradiente de Concentração , Cruzamentos Genéticos , Regulação da Expressão Gênica de Plantas , Genes Dominantes/genética , Genes de Plantas/genética , Estudos de Associação Genética , Immunoblotting , Modelos Biológicos , Mutagênese Insercional/genética , Mutação/genética , Filogenia , Plantas Geneticamente Modificadas , Pirofosfatases/genética , Interferência de RNA , Reprodutibilidade dos Testes , Sementes/metabolismoRESUMO
Malignant hyperthermia (MH) susceptibility is a dominantly inherited disorder in which volatile anesthetics trigger aberrant Ca(2+) release in skeletal muscle and a potentially fatal rise in perioperative body temperature. Mutations causing MH susceptibility have been identified in two proteins critical for excitation-contraction (EC) coupling, the type 1 ryanodine receptor (RyR1) and Ca(V)1.1, the principal subunit of the L-type Ca(2+) channel. All of the mutations that have been characterized previously augment EC coupling and/or increase the rate of L-type Ca(2+) entry. The Ca(V)1.1 mutation R174W associated with MH susceptibility occurs at the innermost basic residue of the IS4 voltage-sensing helix, a residue conserved among all Ca(V) channels [Carpenter D, et al. (2009) BMC Med Genet 10:104-115.]. To define the functional consequences of this mutation, we expressed it in dysgenic (Ca(V)1.1 null) myotubes. Unlike previously described MH-linked mutations in Ca(V)1.1, R174W ablated the L-type current and had no effect on EC coupling. Nonetheless, R174W increased sensitivity of Ca(2+) release to caffeine (used for MH diagnostic in vitro testing) and to volatile anesthetics. Moreover, in Ca(V)1.1 R174W-expressing myotubes, resting myoplasmic Ca(2+) levels were elevated, and sarcoplasmic reticulum (SR) stores were partially depleted, compared with myotubes expressing wild-type Ca(V)1.1. Our results indicate that Ca(V)1.1 functions not only to activate RyR1 during EC coupling, but also to suppress resting RyR1-mediated Ca(2+) leak from the SR, and that perturbation of Ca(V)1.1 negative regulation of RyR1 leak identifies a unique mechanism that can sensitize muscle cells to MH triggers.
Assuntos
Cálcio/metabolismo , Caveolina 1/genética , Acoplamento Excitação-Contração/fisiologia , Predisposição Genética para Doença/genética , Hipertermia Maligna/genética , Músculo Esquelético/metabolismo , Canal de Liberação de Cálcio do Receptor de Rianodina/genética , Análise de Variância , Anestésicos/farmacologia , Temperatura Corporal , Cafeína/farmacologia , DNA Complementar/genética , Acoplamento Excitação-Contração/genética , Fluorescência , Genes Dominantes/genética , Humanos , Microeletrodos , Mutação de Sentido Incorreto/genética , Retículo Sarcoplasmático/metabolismoRESUMO
HISTORY: A 42-year-old man was found to have a four to six fold increase in the level of plasma ferritin since four years. In the age of 10 he had undergone unilateral resection of a dysplastic kidney associated with systemic hypertension. He had also developed recurrent venous thromboses caused by atresia of the inferior vena cava with azygos continuation, known since 23 years. Iron overload or hemochromatosis had been excluded, but despite numerous investigations the exact cause of the hyperferritinemia had not been elucidated. The patient, his grandfather, his mother and a brother had undergone cataract surgeries in both eyes. He presented at admission with prominent veins over the abdomen a postthrombotic syndrome. INVESTIGATION: Laboratory tests revealed a ferritin level 6 times above the upper limit of normal, but iron, transferrin saturation, and transferrin levels were normal. The patient was on oral anticoagulation (INR 2.2). Molecular genetic tests revealed heterozygous mutation IRE+ 32 G > T. DIAGNOSIS, TREATMENT AND COURSE: The findings indicated a hereditary hyperferritinemia cataract syndrome with an autosomal dominant trait. As functions of other organs are not affected, bilateral cataract surgery is "curative". CONCLUSION: Early and correct diagnosis avoids unnecessary diagnostic and therapeutic interventions, such as extended and repeated laboratory tests, liver biopsies, phlebotomies and chelation therapy.
Assuntos
Catarata/congênito , Distúrbios do Metabolismo do Ferro/congênito , Adulto , Catarata/sangue , Catarata/diagnóstico , Catarata/genética , Extração de Catarata , Aberrações Cromossômicas , Análise Mutacional de DNA , Ferritinas/sangue , Genes Dominantes/genética , Triagem de Portadores Genéticos , Testes Genéticos , Humanos , Hipertensão Renal/sangue , Hipertensão Renal/diagnóstico , Hipertensão Renal/genética , Ferro/sangue , Distúrbios do Metabolismo do Ferro/sangue , Distúrbios do Metabolismo do Ferro/diagnóstico , Distúrbios do Metabolismo do Ferro/genética , Proteínas Reguladoras de Ferro/genética , Rim/anormalidades , Masculino , Fenótipo , Síndrome Pós-Trombótica/sangue , Síndrome Pós-Trombótica/diagnóstico , Síndrome Pós-Trombótica/genética , Transferrina/metabolismo , Veia Cava Inferior/anormalidades , Trombose Venosa/sangue , Trombose Venosa/diagnóstico , Trombose Venosa/genéticaRESUMO
⢠A detailed molecular understanding of how oomycete plant pathogens evade disease resistance is essential to inform the deployment of durable resistance (R) genes. ⢠Map-based cloning, transient expression in planta, pathogen transformation and DNA sequence variation across diverse isolates were used to identify and characterize PiAVR2 from potato late blight pathogen Phytophthora infestans. ⢠PiAVR2 is an RXLR-EER effector that is up-regulated during infection, accumulates at the site of haustoria formation, and is recognized inside host cells by potato protein R2. Expression of PiAVR2 in a virulent P. infestans isolate conveys a gain-of-avirulence phenotype, indicating that this is a dominant gene triggering R2-dependent disease resistance. PiAVR2 presence/absence polymorphisms and differential transcription explain virulence on R2 plants. Isolates infecting R2 plants express PiAVR2-like, which evades recognition by R2. PiAVR2 and PiAVR2-like differ in 13 amino acids, eight of which are in the C-terminal effector domain; one or more of these determines recognition by R2. Nevertheless, few polymorphisms were observed within each gene in pathogen isolates, suggesting limited selection pressure for change within PiAVR2 and PiAVR2-like. ⢠Our results direct a search for R genes recognizing PiAVR2-like, which, deployed with R2, may exert strong selection pressure against the P. infestans population.
Assuntos
Phytophthora infestans/patogenicidade , Doenças das Plantas/imunologia , Polimorfismo Genético/genética , Proteínas/metabolismo , Solanum tuberosum/fisiologia , Sequência de Aminoácidos , Mapeamento Cromossômico , Clonagem Molecular , Regulação da Expressão Gênica , Genes Dominantes/genética , Genes de Plantas/genética , Dados de Sequência Molecular , Phytophthora infestans/genética , Phytophthora infestans/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal/genética , Folhas de Planta/genética , Folhas de Planta/imunologia , Folhas de Planta/microbiologia , Folhas de Planta/fisiologia , Estrutura Terciária de Proteína , Proteínas/genética , Solanum/genética , Solanum tuberosum/genética , Solanum tuberosum/imunologia , Solanum tuberosum/microbiologia , Virulência/genética , Fatores de Virulência/genética , Fatores de Virulência/metabolismoAssuntos
Alelos , Brassica/genética , Genes Dominantes/genética , Genes de Plantas/genética , Infertilidade das Plantas/genética , RNA de Plantas/genética , RNA não Traduzido/genética , Brassica/fisiologia , Metilação de DNA/genética , Diploide , Regulação da Expressão Gênica de Plantas/genética , Inativação Gênica , Genes Recessivos/genética , Haplótipos/genética , Infertilidade das Plantas/fisiologia , Proteínas de Plantas/genética , Pólen/genética , Pólen/metabolismo , Reprodução/genética , Reprodução/fisiologiaRESUMO
A diploid organism has two copies of each gene, one inherited from each parent. The expression of two inherited alleles is sometimes biased by the effects known as dominant/recessive relationships, which determine the final phenotype of the organism. To explore the mechanisms underlying these relationships, we have examined the monoallelic expression of S-locus protein 11 genes (SP11), which encode the male determinants of self-incompatibility in Brassica. We previously reported that SP11 expression was monoallelic in some S heterozygotes, and that the promoter regions of recessive SP11 alleles were specifically methylated in the anther tapetum. Here we show that this methylation is controlled by trans-acting small non-coding RNA (sRNA). We identified inverted genomic sequences that were similar to the recessive SP11 promoters in the flanking regions of dominant SP11 alleles. These sequences were specifically expressed in the anther tapetum and processed into 24-nucleotide sRNA, named SP11 methylation inducer (Smi). Introduction of the Smi genomic region into the recessive S homozygotes triggered the methylation of the promoter of recessive SP11 alleles and repressed their transcription. This is an example showing sRNA encoded in the flanking region of a dominant allele acts in trans to induce transcriptional silencing of the recessive allele. Our finding may provide new insights into the widespread monoallelic gene expression systems.
Assuntos
Alelos , Brassica/genética , Inativação Gênica , Genes Dominantes/genética , Genes de Plantas/genética , Infertilidade das Plantas/genética , RNA de Plantas/genética , RNA não Traduzido/genética , Sequência de Bases , Brassica/fisiologia , Metilação de DNA , Diploide , Flores/genética , Regulação da Expressão Gênica de Plantas/genética , Genes Recessivos/genética , Haplótipos/genética , Heterozigoto , Homozigoto , Dados de Sequência Molecular , Fenótipo , Infertilidade das Plantas/fisiologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Pólen/genética , Pólen/metabolismo , Polinização/genética , Regiões Promotoras Genéticas/genética , Reprodução/genética , Reprodução/fisiologia , Transcrição Gênica/genética , Transgenes/genéticaRESUMO
Familial idiopathic basal ganglia calcification (FIBGC) is an inherited neurodegenerative disorder characterized by the accumulation of calcium deposits in different brain regions, particularly in the basal ganglia. FIBGC usually follows an autosomal dominant pattern of inheritance. Despite the mapping to chromosome 14q of a susceptibility locus for IBGC (IBCG1) in one family, this locus has been excluded in several others, demonstrating genetic heterogeneity in this disorder. The etiology of this disorder thus remains largely unknown. Using a large extended multigenerational Italian family from South Tyrol with 17 affected in a total of 56 members, we performed a genome-wide linkage analysis in which we were able to exclude linkage to the IBCG1 locus on chromosome 14q and obtain evidence of a novel locus on chromosome 2q37. Electronic supplementary material. The online version of this article (doi:10.1007/s12031-009-9287-3) contains supplementary material, which is available to authorized users.
Assuntos
Doenças dos Gânglios da Base/genética , Calcinose/genética , Cromossomos Humanos Par 2/genética , Loci Gênicos/genética , Predisposição Genética para Doença/genética , Mutação/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Gânglios da Base/metabolismo , Gânglios da Base/patologia , Gânglios da Base/fisiopatologia , Doenças dos Gânglios da Base/metabolismo , Doenças dos Gânglios da Base/fisiopatologia , Calcinose/metabolismo , Calcinose/fisiopatologia , Transtornos Cromossômicos/genética , Mapeamento Cromossômico , Cromossomos Humanos Par 14/genética , Análise Mutacional de DNA , Feminino , Genes Dominantes/genética , Ligação Genética/genética , Marcadores Genéticos/genética , Testes Genéticos , Variação Genética/genética , Humanos , Padrões de Herança/genética , Itália , Masculino , Pessoa de Meia-Idade , LinhagemRESUMO
BACKGROUND: Autosomal dominant partial epilepsy with auditory features (ADPEAF) is an idiopathic focal epilepsy syndrome with auditory symptoms or receptive aphasia as major ictal manifestations, frequently associated with mutations in the leucine-rich, glioma inactivated 1 (LGI1) gene. Although affected subjects do not have structural abnormalities detected on routine MRI, a lateral temporal malformation was identified through high resolution MRI in one family. We attempted to replicate this finding and to assess auditory and language processing in ADPEAF using fMRI and magnetoencephalography (MEG). METHODS: We studied 17 subjects (10 affected mutation carriers, 3 unaffected carriers, 4 noncarriers) in 7 ADPEAF families, each of which had a different LGI1 mutation. Subjects underwent high-resolution structural MRI, fMRI with an auditory description decision task (ADDT) and a tone discrimination task, and MEG. A control group comprising 26 volunteers was also included. RESULTS: We found no evidence of structural abnormalities in any of the 17 subjects. On fMRI with ADDT, subjects with epilepsy had significantly less activation than controls. On MEG with auditory stimuli, peak 2 auditory evoked field latency was significantly delayed in affected individuals compared to controls. CONCLUSIONS: These findings do not support the previous report of a lateral temporal malformation in autosomal dominant partial epilepsy with auditory features (ADPEAF). However, our fMRI and magnetoencephalography data suggest that individuals with ADPEAF have functional impairment in language processing.
Assuntos
Córtex Auditivo/fisiopatologia , Percepção Auditiva/genética , Epilepsias Parciais/complicações , Transtornos da Linguagem/fisiopatologia , Percepção da Fala/genética , Estimulação Acústica , Adulto , Córtex Auditivo/patologia , Mapeamento Encefálico , Transtornos Cromossômicos/complicações , Transtornos Cromossômicos/genética , Epilepsias Parciais/genética , Feminino , Lateralidade Funcional/genética , Genes Dominantes/genética , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Transtornos da Linguagem/genética , Transtornos da Linguagem/patologia , Testes de Linguagem , Imageamento por Ressonância Magnética , Magnetoencefalografia , Masculino , Mutação/genética , Proteínas/genética , Tempo de Reação/genéticaRESUMO
Fundamento: La distinción entre el hiperinsulinismo congénito (CHI) focal y difuso es esencial de cara al tratamiento y pronóstico de la enfermedad. El objetivo es presentar el primer caso de CHI focal diagnosticado en España combinando los estudios genético y PET-TC. Métodos: Paciente de 13 meses con CHI y pruebas de imagen convencionales normales, tratado con diazóxido, control dietético y alimentación por gastrostomía. Se analizó la secuencia de los genes ABCC8 y KCNJ11, y realizó una PET-TC con 18F-fluoro-L-DOPA. Resultados: Se detectó una mutación patogénica (G111R) en el alelo paterno de ABCC8. La PET-TC demostró un foco hipercaptante en el cuerpo del páncreas compatible con un adenoma confirmado histopatológicamente. Tras la cirugía el paciente continúa asintomático sin tratamiento farmacológico ni medidas dietéticas. Conclusiones: La combinación del análisis genético y la PET-TC con 18F-fluoro-L-DOPA muestra un gran potencial para la identificación, localización y guía de la cirugía del CHI (AU)
Background: Congenital hyperinsulinism (CHI) is the most common cause of persistent hypoglycaemia in infancy. The differential diagnosis between focal and diffuse forms of CHI is of great importance when planning surgery. The aim of this article is to show the first case of focal CHI diagnosed in Spain using PET-CT imaging combined with genetic analysis. Methods: A 13 month child with CHI and normal conventional radiological investigations treated with diazoxide, diet control and feeding by gastrostomy is presented. Genetic analysis of ABCC8 and KCNJ11 genes and PET-TAC using 18F-fluoro-L-DOPA were performed. Results: A pathological mutation (G111R) in the paternal allele of ABCC8 was detected. PET-CT scanning using 18F-fluoro-L-DOPA showed a focus of high uptake in the body of the pancreas compatible with adenoma that was hystopathologically confirmed. After surgical resection the patient is asymptomatic without needing either pharmacological treatment or dietetic control. Conclusions: The combination of genetic analysis and 18F-fluoro-L-DOPA PET-TAC shows a great potential for the identification, location and guideline for surgery in CHI (AU)
Assuntos
Humanos , Masculino , Lactente , Hiperinsulinismo/diagnóstico , Hiperinsulinismo/genética , Prognóstico Clínico Dinâmico Homeopático , Hipoglicemia/complicações , Hidrocortisona/uso terapêutico , Glucagon/uso terapêutico , Octreotida/uso terapêutico , Genes Dominantes , Genes Dominantes/genética , Mutação/genética , Gastrostomia , Diazóxido/uso terapêutico , Consentimento Livre e Esclarecido , Pâncreas/anormalidades , Pâncreas , Aberrações Cromossômicas/embriologia , Transtornos Cromossômicos/complicações , Genes Dominantes/fisiologia , Expressão Gênica/fisiologiaRESUMO
PURPOSE OF REVIEW: While epilepsy describes a heterogeneous array of syndromes, the conventional view is that there is a common underlying failure in the ability of GABAergic inhibition to overcome excessive synaptic excitation. This review explores the possibility that enhanced GABAergic inhibition in the neocortex could also be proepileptogenic. RECENT FINDINGS: Recently, two mouse strains carrying mutant alleles of the alpha4 subunit of the nicotinic acetylcholine receptor that are associated with autosomal dominant nocturnal frontal lobe epilepsy have been found to show spontaneous seizures. Recordings from neocortical pyramidal neurons in vitro show that the autosomal dominant nocturnal frontal lobe epilepsy mutations are associated with large selective increases in nicotine-evoked GABAergic inhibition, which may be key factor in epileptogenesis, as the seizures in vivo are blocked by subconvulsive doses of the GABAA receptor antagonist, picrotoxin. SUMMARY: The precise links between the observed gain of neocortical inhibition and development of seizures in autosomal dominant nocturnal frontal lobe epilepsy mice remain unknown. Recent insights into the functional properties of cortical GABAergic circuits, however, suggest several possible pathways to be explored, whose elucidation could enable selective therapeutic interventions.