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Prótesis Osicular , Otosclerosis/cirugía , Implantación de Prótesis/métodos , Cirugía del Estribo/métodos , Adulto , Aleaciones , Femenino , Audición , Humanos , Masculino , Persona de Mediana Edad , Diseño de Prótesis , Implantación de Prótesis/instrumentación , Estudios Retrospectivos , Cirugía del Estribo/instrumentación , Resultado del TratamientoRESUMEN
BACKGROUND: Multiple sclerosis (MS) may affect other cranial nerves besides the optic nerve. Sudden sensorineural hearing loss (SSHL), possibly caused by a deficit in the auditory tract, including the vestibulocochlear nerve, is sometimes associated with MS. OBJECTIVES: We aimed to assess the incidence of SSHL among MS patients, its frequency as an initial symptom of MS, and magnetic resonance imaging (MRI) findings associated with SSHL in MS. METHODS: We collected retrospectively all patients diagnosed with MS and SSHL at the Helsinki University Hospital between 2004 and 2014. Patients with both diagnoses were re-evaluated using hospital medical records, audiograms and head MRI scans. RESULTS: A total of 2736 patients were diagnosed with MS, 1581 patients with SSHL, and 18 patients (0.7% of all MS patients) with both; two patients presented with SSHL as an initial symptom of MS. The annual incidence of SSHL was 59.8/100 000 (95% confidence interval (CI) 37.7-94.9) in MS patients, and 12.4/100 000 (95% CI 11.8-13.0) in the normal population. CONCLUSION: SSHL is a rare symptom of MS and is even less frequent as an initial symptom. Its incidence in MS patients, however, markedly exceeds that in the normal population.
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Ablación por Catéter , Pabellón Auricular , Queloide/cirugía , Adolescente , Adulto , Anciano , Perforación del Cuerpo/efectos adversos , Niño , Estudios de Cohortes , Femenino , Humanos , Queloide/etiología , Queloide/patología , Masculino , Persona de Mediana Edad , Resultado del Tratamiento , Adulto JovenRESUMEN
Agonists of the allosteric benzodiazepine site of GABAA receptors bind at the interface of the alpha and gamma subunits. Here, we tested the in vivo contribution of the gamma2 subunit to the actions of zolpidem, an alpha1 subunit selective benzodiazepine agonist, by generating mice with a phenylalanine (F) to isoleucine (I) substitution at position 77 in the gamma2 subunit. The gamma2F77I mutation has no major effect on the expression of GABAA receptor subunits in the cerebellum. The potency of zolpidem, but not that of flurazepam, for the inhibition of [3H]flunitrazepam binding to cerebellar membranes is greatly reduced in gamma2I77/I77 mice. Zolpidem (1 microM) increased both the amplitude and decay of miniature inhibitory postsynaptic currents (mIPSCs) in Purkinje cells of control C57BL/6 (34% and 92%, respectively) and gamma2F77/F77 (20% and 84%) mice, but not in those of gamma2F77I mice. Zolpidem tartrate had no effect on exploratory activity (staircase test) or motor performance (rotarod test) in gamma2I77/I77 mice at doses up to 30 mg/kg (i.p.) that strongly sedated or impaired the control mice. Flurazepam was equally effective in enhancing mIPSCs and disrupting performance in the rotarod test in control and gamma2I77/I77 mice. These results show that the effect of zolpidem, but not flurazepam, is selectively eliminated in the brain by the gamma2F77I point mutation.
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Agonistas del GABA/farmacología , Mutación Puntual , Piridinas/farmacología , Receptores de GABA-A/genética , Sustitución de Aminoácidos , Animales , Secuencia de Bases , Cartilla de ADN , Flunitrazepam/farmacocinética , Ratones , Ratones Mutantes , Polimorfismo de Nucleótido Simple/genética , ZolpidemRESUMEN
Angelman syndrome is a severe neurodevelopmental disorder with cognitive impairment and neurological deficits. It results from a maternal deletion of human chromosome 15q11-13 containing two candidate genes E6-P ubiquitin-protein ligase (UBE3A) and GABA(A) receptor beta3 subunit (GABRB3), the latter of which has been also linked to autism. To clarify the potential role of GABA(A) beta3 subunit-containing inhibitory receptors in these disorders, we applied ligand autoradiography on brain sections from mice with inactivated GABRB3 or maternal UBE3A genes. Binding of GABA(A) receptor channel ([(35)S]t-butylbicyclophosphorothionate) and benzodiazepine ([(3)H]Ro 15-4513) site ligands was reduced in selected brain regions of the beta3-deficient mice as compared to controls, while the UBE3A-deficient mice failed to show reduced GABA(A) receptors. The results, suggesting two different pathophysiological mechanisms, are in agreement with positron emission tomography results from Angelman syndrome patients of the corresponding genetic backgrounds.
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Síndrome de Angelman/metabolismo , Encéfalo/metabolismo , Modelos Animales de Enfermedad , Receptores de GABA-A/metabolismo , Síndrome de Angelman/genética , Animales , Femenino , Ligasas/deficiencia , Ligasas/genética , Ligasas/metabolismo , Masculino , Ratones , Ratones Mutantes , Unión Proteica/fisiología , Receptores de GABA-A/deficiencia , Receptores de GABA-A/genética , Ubiquitina-Proteína LigasasRESUMEN
We generated transgenic (Thy1alpha6) mice in which the GABA(A) receptor alpha6 subunit, whose expression is usually confined to granule cells of cerebellum and cochlear nuclei, is ectopically expressed under the control of the pan-neuronal Thy-1.2 promoter. Strong Thy1alpha6 subunit expression occurs, for example, in deep cerebellar nuclei, layer V iscocortical and hippocampal pyramidal cells and dentate granule cells. Ligand binding and protein biochemistry show that most forebrain alpha6 subunits assemble as alpha6betagamma2-type receptors, and some as alpha1alpha6betagamma2 and alpha3alpha6betagamma2 receptors. Electron microscopic immunogold labeling shows that most Thy1-derived alpha6 immunoreactivity is in the extrasynaptic plasma membrane of dendrites and spines in both layer V isocortical and CA1pyramidal cells. Synaptic immunolabeling is rare. Consistent with the alpha6 subunits' extrasynaptic localization, Thy1alpha6 CA1 pyramidal neurons have a five-fold increased tonic GABA(A) receptor-mediated current compared with wild-type cells; however, the spontaneous IPSC frequency and the mIPSC amplitude in Thy1alpha6 mice decrease 37 and 30%, respectively compared with wild-type. Our results strengthen the idea that GABA(A) receptors containing alpha6 subunits can function as extrasynaptic receptors responsible for tonic inhibition and further suggest that a homeostatic mechanism might operate, whereby increased tonic inhibition causes a compensatory decrease in synaptic GABA(A) receptor responses.
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Hipocampo/metabolismo , Células Piramidales/metabolismo , Receptores de GABA-A/biosíntesis , Potenciales de Acción/fisiología , Secuencia de Aminoácidos , Animales , Autorradiografía , Electrofisiología , Colorantes Fluorescentes , Congelación , Hipocampo/citología , Inmunohistoquímica , Hibridación in Situ , Ligandos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Microscopía Electrónica , Datos de Secuencia Molecular , Prosencéfalo/metabolismo , Fracciones Subcelulares/metabolismo , Sinapsis/efectos de los fármacos , Adhesión del TejidoRESUMEN
A GABA(A) receptor delta subunit-deficient mouse line was created by homologous recombination in embryonic stem cells to investigate the role of the subunit in the brain GABA(A) receptors. High-affinity [(3)H]muscimol binding to GABA sites as studied by ligand autoradiography was reduced in various brain regions of delta(-/-) animals. [(3)H]Ro 15-4513 binding to benzodiazepine sites was increased in delta(-/-) animals, partly due to an increment of diazepam-insensitive receptors, indicating an augmented forebrain assembly of gamma 2 subunits with alpha 4 subunits. In the western blots of forebrain membranes of delta(-/-) animals, the level of gamma 2 subunit was increased and that of alpha 4 decreased, while the level of alpha1 subunits remained unchanged. In the delta(-/-) forebrains, the remaining alpha 4 subunits were associated more often with gamma 2 subunits, since there was an increase in the alpha 4 subunit level immunoprecipitated by the gamma 2 subunit antibody. The pharmacological properties of t-butylbicyclophosphoro[(35)S]thionate binding to the integral ion-channel sites were slightly altered in the forebrain and cerebellum, consistent with elevated levels of alpha 4 gamma 2 and alpha 6 gamma 2 subunit-containing receptors, respectively.The altered pharmacology of forebrain GABA(A) receptors and the decrease of the alpha 4 subunit level in delta subunit-deficient mice suggest that the delta subunit preferentially assembles with the alpha 4 subunit. The delta subunit seems to interfere with the co-assembly of alpha 4 and gamma 2 subunits and, therefore, in its absence, the gamma 2 subunit is recruited into a larger population of alpha 4 subunit-containing functional receptors. These results support the idea of subunit competition during the assembly of native GABA(A) receptors.
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Encéfalo/metabolismo , Inhibición Neural/fisiología , Neuronas/metabolismo , Receptores de GABA-A/deficiencia , Transmisión Sináptica/fisiología , Ácido gamma-Aminobutírico/metabolismo , Marcadores de Afinidad , Animales , Azidas , Benzodiazepinas/agonistas , Sitios de Unión/efectos de los fármacos , Sitios de Unión/fisiología , Encéfalo/efectos de los fármacos , Femenino , Agonistas del GABA , Canales Iónicos/efectos de los fármacos , Canales Iónicos/genética , Canales Iónicos/metabolismo , Masculino , Ratones , Ratones Noqueados , Muscimol , Mutación/genética , Inhibición Neural/efectos de los fármacos , Neuronas/efectos de los fármacos , Ensayo de Unión Radioligante , Receptores de GABA-A/efectos de los fármacos , Receptores de GABA-A/genética , Transmisión Sináptica/efectos de los fármacos , Tritio , Ácido gamma-Aminobutírico/farmacologíaRESUMEN
An increase in the activity of brain stem locus coeruleus noradrenergic neurons has been hypothesised to be a major factor accounting for opiate withdrawal symptoms. These neurons are under GABAergic inhibition. Their GABA(A) receptors have unique pharmacological properties, most likely due to the enriched expression of GABA(A) receptor subtypes containing novel epsilon and straight theta subunits. Using in situ hybridisation of cryostat sections, we now report a significant increase in the epsilon subunit mRNA expression after precipitation of opioid withdrawal by naloxone. Similar changes were detected in tyrosine hydroxylase mRNA expression. The results suggest increased formation of unique GABA(A) receptor subtype(s) in the locus coeruleus neurons during increased neuronal activity.
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Expresión Génica/efectos de los fármacos , Locus Coeruleus/efectos de los fármacos , Morfina/farmacología , Neuronas/efectos de los fármacos , ARN Mensajero/efectos de los fármacos , Receptores de GABA-A/genética , Síndrome de Abstinencia a Sustancias/metabolismo , Animales , Interacciones Farmacológicas/fisiología , Expresión Génica/fisiología , Locus Coeruleus/metabolismo , Masculino , Dependencia de Morfina/tratamiento farmacológico , Dependencia de Morfina/metabolismo , Dependencia de Morfina/fisiopatología , Naloxona/farmacología , Antagonistas de Narcóticos/farmacología , Neuronas/metabolismo , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Síndrome de Abstinencia a Sustancias/tratamiento farmacológico , Síndrome de Abstinencia a Sustancias/fisiopatología , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Gamma-ainobutyric acid type A (GABA(A)) receptor ionophore ligand t-[35S]butylbicyclophosphorothionate ([35S]TBPS) was used in an autoradiographic assay on brain cryostat sections to visualize and characterize atypical GABA-insensitive [35S]TBPS binding previously described in certain recombinant GABA(A) receptors and the cerebellar granule cell layer. Picrotoxinin-sensitive but 1-mM GABA-insensitive [35S]TBPS binding was present in the rat cerebellar granule cell layer, many thalamic nuclei, subiculum and the internal rim of the cerebral cortex, amounting in these regions up to 6% of the basal binding determined in the absence of exogenous GABA. Similar binding properties were detected also in human and chicken brain sections. Like the GABA-sensitive [35S]TBPS binding, GABA-insensitive binding was profoundly decreased by pentobarbital, pregnanolone, loreclezole and Mg2+. The binding was reversible and apparently dependent on Cl- ions. Localization of the GABA-insensitive [35S]TBPS binding was not identical to that of high-affinity [3H]muscimol binding and diazepam-insensitive [3H]Ro 15-4513 binding, two previously established receptor subtype-dependent binding heterogeneities in the rat brain. The present study reveals a component of the GABA-ionophore enriched in the thalamus and cerebellar granule cells, possibly representing poorly desensitized or desensitizing receptors.
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Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , Cerebelo/metabolismo , Convulsivantes/farmacología , Agonistas del GABA/farmacología , Muscimol/farmacología , Receptores de GABA-A/metabolismo , Tálamo/metabolismo , Marcadores de Afinidad/metabolismo , Marcadores de Afinidad/farmacología , Animales , Azidas/metabolismo , Azidas/farmacología , Benzodiazepinas/metabolismo , Benzodiazepinas/farmacología , Unión Competitiva , Compuestos Bicíclicos Heterocíclicos con Puentes/metabolismo , Pollos , Convulsivantes/metabolismo , Agonistas del GABA/metabolismo , Humanos , Masculino , Muscimol/metabolismo , Picrotoxina/análogos & derivados , Picrotoxina/farmacología , Ensayo de Unión Radioligante , Ratas , Ratas Wistar , Sesterterpenos , Radioisótopos de Azufre , Tritio , Ácido gamma-Aminobutírico/farmacologíaRESUMEN
We searched for subunit correlations for GABA(A) receptor-associated atypically GABA-insensitive [35S]TBPS binding. The homomeric beta3 subunit receptors could be excluded, as GABA-insensitive [35S]TBPS binding was present in beta3-/- mice. Localization of GABA-insensitive [35S]TBPS binding correlated best with those of delta, alpha4 and alpha6 subunit mRNAs. The amounts of GABA-insensitive [35S]TBPS binding components were increased in delta-/- mice, but dramatically reduced in alpha6-/- mice, suggesting a role for alpha6 but excluding delta subunits.
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Química Encefálica/genética , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , Convulsivantes/farmacología , Receptores de GABA-A , Animales , Sitios de Unión , Compuestos Bicíclicos Heterocíclicos con Puentes/metabolismo , Convulsivantes/metabolismo , Agonistas del GABA/metabolismo , Agonistas del GABA/farmacología , Ratones , Ratones Noqueados , Datos de Secuencia Molecular , ARN Mensajero/análisis , Ensayo de Unión Radioligante , Receptores de GABA-A/química , Receptores de GABA-A/genética , Receptores de GABA-A/metabolismo , Radioisótopos de AzufreRESUMEN
A deamination product of histidine, urocanic acid, accumulates in the skin of mammals as trans-urocanic acid. Ultraviolet (UV) irradition converts it to the cis-isomer that is an important mediator in UV-induced immunosuppression. We have recently shown that urocanic acid interferes with the agonist binding to GABA(A) receptors. We now report that the effects of urocanic acid on binding of a convulsant ligand (t-butylbicyclo[35S]phosphorothionate) to GABA(A) receptors in brain membrane homogenates are dependent on pH of the incubation medium, the agonistic actions being enhanced at the normal pH of the skin (5.5). Using Xenopus laevis oocytes expressing recombinant rat alpha1beta1gamma2S GABA(A) receptors, the low pH potentiated the direct agonistic action of trans-urocanic acid under two-electrode voltage-clamp, whereas cis-urocanic acid retained its low efficacy both at pH 5.5 and 7.4. The results thus indicate clear differences between urocanic acid isomers in functional activity at one putative receptor site of immunosuppression, the GABA(A) receptor, the presence of which in the skin remains to be demonstrated.
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Moduladores del GABA/farmacología , Receptores de GABA-A/efectos de los fármacos , Ácido Urocánico/farmacología , Animales , Compuestos Bicíclicos Heterocíclicos con Puentes/metabolismo , Relación Dosis-Respuesta a Droga , Concentración de Iones de Hidrógeno , Tolerancia Inmunológica/efectos de la radiación , Masculino , Ratas , Ratas Wistar , Receptores de GABA-A/fisiología , Estereoisomerismo , Rayos UltravioletaRESUMEN
Clustering of GABA(A) receptor alpha1, alpha6, beta2, and gamma2 subunit genes on mouse chromosome 11/human chromosome 5 may have functional significance for coordinating expression patterns, but until now there has been no evidence for cross-talk between the genes. However, altering the structure of the alpha6 gene, specifically expressed in the cerebellum, with neomycin gene insertions in two different experiments unexpectedly reduced the expression of the widespread alpha1 and beta2 genes in the forebrain. There were corresponding reductions in the levels of alpha1 and beta2 subunit proteins and in autoradiographic ligand binding densities to GABA(A) receptors in the forebrain of alpha6-/- mice. The gamma2 mRNA level was not changed, nor were beta3 and delta mRNAs. The data suggest that elements in the neo gene may have an influence over long distances in the GABA(A) subunit gene complex on as yet undefined structures coordinating the expression of the alpha1 and beta2 genes.
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Regulación de la Expresión Génica , Marcación de Gen , Familia de Multigenes/genética , Prosencéfalo/metabolismo , Receptores de GABA-A/genética , Receptores de GABA-A/metabolismo , Animales , Autorradiografía , Western Blotting , Cerebelo/metabolismo , Genes Reporteros , Humanos , Ligandos , Ratones , Ratones Noqueados , ARN Mensajero/biosíntesis , Ensayo de Unión RadioliganteRESUMEN
Previously, GABA(A) receptor epsilon and theta subunits have been identified only in human. Here, we describe properties of the epsilon and theta subunit genes from mouse and rat that reveal an unusually high level of divergence from their human homologs. In addition to a low level of amino acid sequence conservation ( approximately 70%), the rodent epsilon subunit cDNAs encode a unique Pro/Glx motif of approximately 400 residues within the N-terminal extracellular domain of the subunits. Transcripts of the rat epsilon subunit were detected in brain and heart, whereas the mouse theta subunit mRNA was detectable in brain, lung, and spleen by Northern blot analysis. In situ hybridization revealed a particularly strong signal for both subunit mRNAs in rat locus ceruleus in which expression was detectable from the first postnatal day. Lower levels of coexpression were also detected in other brainstem nuclei and in the hypothalamus. However, the expression pattern of theta subunit mRNA was more widespread than that of epsilon subunit, being found also in the cerebral cortex of rat pups. In contrast to primate brain, neither subunit was expressed in the hippocampus or substantia nigra. The results indicate that GABA(A) receptor epsilon and theta subunits are evolving at a much faster rate than other known GABA(A) receptor subunits and that their expression patterns and functional properties may differ significantly between species.