RESUMEN
Early identification of Alzheimer's disease (AD) risk factors would aid development of interventions to delay the onset of dementia, but current biomarkers are invasive and/or costly to assess. Validated plasma biomarkers would circumvent these challenges. We previously identified the kinase DYRK1A in plasma. To validate DYRK1A as a biomarker for AD diagnosis, we assessed the levels of DYRK1A and the related markers brain-derived neurotrophic factor (BDNF) and homocysteine in two unrelated AD patient cohorts with age-matched controls. Receiver-operating characteristic curves and logistic regression analyses showed that combined assessment of DYRK1A, BDNF and homocysteine has a sensitivity of 0.952, a specificity of 0.889 and an accuracy of 0.933 in testing for AD. The blood levels of these markers provide a diagnosis assessment profile. Combined assessment of these three markers outperforms most of the previous markers and could become a useful substitute to the current panel of AD biomarkers. These results associate a decreased level of DYRK1A with AD and challenge the use of DYRK1A inhibitors in peripheral tissues as treatment. These measures will be useful for diagnosis purposes.
Asunto(s)
Enfermedad de Alzheimer/sangre , Factor Neurotrófico Derivado del Encéfalo/sangre , Homocisteína/sangre , Proteínas Serina-Treonina Quinasas/sangre , Proteínas Tirosina Quinasas/sangre , Anciano , Enfermedad de Alzheimer/diagnóstico , Enfermedad de Alzheimer/inmunología , Animales , Anticuerpos Monoclonales/metabolismo , Anticuerpos Monoclonales/farmacología , Biomarcadores/sangre , Femenino , Humanos , Masculino , Ratones Endogámicos C57BL , Ratones Transgénicos , Persona de Mediana Edad , Proteínas Serina-Treonina Quinasas/inmunología , Proteínas Tirosina Quinasas/inmunología , Curva ROC , Quinasas DyrKRESUMEN
To determine whether apparent involvement of DYRK1A in Alzheimer's disease (AD) pathology makes it a candidate plasma biomarker for diagnosis, we developed a method to quantify plasma DYRK1A by immunoblot in transgenic mouse models having different gene dosages of Dyrk1a, and, consequently, different relative protein expression. Then, we measured plasma DYRK1A levels in 26 patients with biologically confirmed AD and 25 controls (negative amyloid imaging available on 13). DYRK1A was detected in transgenic mouse brain and plasma samples, and relative levels of DYRK1A correlated with the gene copy number. In plasma from AD patients, DYRK1A levels were significantly lower compared with controls (P<0.0001). Results were similar when we compared AD patients with the subgroup of controls confirmed by negative amyloid imaging. In a subgroup of patients with early AD (CDR=0.5), lower DYRK1A expression was confirmed. In contrast, no difference was found in levels of DYRK1B, the closest relative of DYRK1A, between AD patients and controls. Further, AD patients exhibited a positive correlation between plasma DYRK1A levels and cerebrospinal fluid tau and phosphorylated-tau proteins, but no correlation with amyloid-ß42 levels and Pittsburgh compound B cortical binding. DYRK1A levels detected in lymphoblastoid cell lines from AD patients were also lower when compared with cells from age-matched controls. These findings suggest that reduced DYRK1A expression might be a novel plasma risk factor for AD.
Asunto(s)
Enfermedad de Alzheimer/sangre , Enfermedad de Alzheimer/genética , Biomarcadores/sangre , Marcadores Genéticos/genética , Proteínas Serina-Treonina Quinasas/sangre , Proteínas Serina-Treonina Quinasas/genética , Proteínas Tirosina Quinasas/sangre , Proteínas Tirosina Quinasas/genética , Anciano , Enfermedad de Alzheimer/diagnóstico , Animales , Encéfalo/metabolismo , Encéfalo/patología , Modelos Animales de Enfermedad , Femenino , Estudios de Asociación Genética , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Persona de Mediana Edad , Tomografía de Emisión de Positrones , Valor Predictivo de las Pruebas , Quinasas DyrKRESUMEN
In the brain, programmed cell death (PCD) serves to adjust the numbers of the different types of neurons during development, and its pathological reactivation in the adult leads to neurodegeneration. Dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 1A (DYRK1A) is a pleiotropic kinase involved in neural proliferation and cell death, and its role during brain growth is evolutionarily conserved. Human DYRK1A lies in the Down syndrome critical region on chromosome 21, and heterozygous mutations in the gene cause microcephaly and neurological dysfunction. The mouse model for DYRK1A haploinsufficiency (the Dyrk1a(+/-) mouse) presents neuronal deficits in specific regions of the adult brain, including the substantia nigra (SN), although the mechanisms underlying these pathogenic effects remain unclear. Here we study the effect of DYRK1A copy number variation on dopaminergic cell homeostasis. We show that mesencephalic DA (mDA) neurons are generated in the embryo at normal rates in the Dyrk1a haploinsufficient model and in a model (the mBACtgDyrk1a mouse) that carries three copies of Dyrk1a. We also show that the number of mDA cells diminishes in postnatal Dyrk1a(+/-) mice and increases in mBACtgDyrk1a mice due to an abnormal activity of the mitochondrial caspase9 (Casp9)-dependent apoptotic pathway during the main wave of PCD that affects these neurons. In addition, we show that the cell death induced by 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP), a toxin that activates Casp9-dependent apoptosis in mDA neurons, is attenuated in adult mBACtgDyrk1a mice, leading to an increased survival of SN DA neurons 21 days after MPTP intoxication. Finally, we present data indicating that Dyrk1a phosphorylation of Casp9 at the Thr125 residue is the mechanism by which this kinase hinders both physiological and pathological PCD in mDA neurons. These data provide new insight into the mechanisms that control cell death in brain DA neurons and they show that deregulation of developmental apoptosis may contribute to the phenotype of patients with imbalanced DYRK1A gene dosage.
Asunto(s)
Apoptosis , Neuronas Dopaminérgicas/metabolismo , Intoxicación por MPTP/metabolismo , Mesencéfalo/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Tirosina Quinasas/metabolismo , Animales , Caspasa 9/genética , Caspasa 9/metabolismo , Supervivencia Celular/genética , Modelos Animales de Enfermedad , Neuronas Dopaminérgicas/patología , Humanos , Intoxicación por MPTP/genética , Intoxicación por MPTP/patología , Mesencéfalo/patología , Ratones , Ratones Noqueados , Proteínas Serina-Treonina Quinasas/genética , Proteínas Tirosina Quinasas/genética , Quinasas DyrKRESUMEN
This study was designed to determine whether Coxsackie adenovirus receptor (CAR) and alpha nu beta3/alpha nu beta5 integrin co-receptors are involved in adenovirus gene transfer in the rat cochlea. We find that CAR and integrin co-receptors are expressed in every cell subtype transduced by the adenoviral vector Ad5 DeltaE1-E3/cytomegalovirus/green fluorescent protein (GFP) on cochlear slices in vitro. The spiral ganglion neurons, which do not express CAR, were not transduced by the virus. Blocking these receptors by monoclonal antibodies decreased transgene expression, whereas disrupting tight junctions with ethylenediaminetetraacetic acid led to an increased transgene expression. However, sensory hair cells and strial cells also expressing CAR and alpha nu integrins were not transduced by the vector. GFP expression was also studied in vivo. Perilymphatic perfusion of adenovirus in vivo did not affect hearing and only cells lining the perilymphatic spaces were transduced. Endolymphatic perfusion resulted in low-frequency hearing loss and although some cells of the organ of Corti were efficiently transduced, the sensory and the strial cells were not. Transduced sensory and strial cells were occasionally observed in cochleas after single shot of adenovirus. Pretreatment with anti-CAR and anti-alpha nu antibodies decreases GFP expression in vivo, suggesting that the CAR/alpha nu integrin pathway is involved in adenovirus transduction in the cochlea.
Asunto(s)
Adenoviridae/genética , Cóclea/metabolismo , Vectores Genéticos/administración & dosificación , Integrinas/metabolismo , Receptores Virales/metabolismo , Transducción Genética/métodos , Potenciales de Acción , Animales , Cóclea/virología , Nervio Coclear/fisiología , Proteína de la Membrana Similar al Receptor de Coxsackie y Adenovirus , Expresión Génica , Terapia Genética/métodos , Vectores Genéticos/genética , Proteínas Fluorescentes Verdes/análisis , Proteínas Fluorescentes Verdes/genética , Humanos , Inmunohistoquímica , Inyecciones , Integrina alfa5/análisis , Integrina alfa5/metabolismo , Cadenas beta de Integrinas/análisis , Cadenas beta de Integrinas/metabolismo , Integrina beta3/análisis , Integrina beta3/metabolismo , Integrinas/análisis , Microscopía Fluorescente , Modelos Animales , Ratas , Ratas Wistar , Técnicas de Cultivo de Tejidos , TransgenesRESUMEN
The dual-specificity tyrosine-phosphorylated and regulated kinase 1A (DYRK1A) gene encodes a protein kinase known to play a critical role in neurodevelopment. Mice with one functional copy of Dyrk1A (Dyrk1A(+/-)) display a marked hypoactivity and altered gait dynamics in basal conditions and in novel environments. Dopamine (DA) is a key neurotransmitter in motor behavior and genetic deletion of certain genes directly related to the dopaminergic system has a strong impact on motor activity. We have studied the effects of reduced Dyrk1A expression on the function of the nigrostriatal dopaminergic system. To characterize the dopaminergic system in DYRK1A(+/-) mice, we have used behavioral, pharmacological, histological, neurochemical and neuroimaging (microPET) techniques in a multidisciplinary approach. Dyrk1A(+/-) mice exhibited decreased striatal DA levels, reduced number of DA neurons in the substantia nigra pars compacta, as well as altered behavioral responses to dopaminergic agents. Moreover, microdialysis experiments revealed attenuated striatal DA release and positron emission tomography scan display reduced forebrain activation when challenged with amphetamine, in Dyrk1A(+/-) compared with wild-type mice. These data indicate that Dyrk1A is essential for a proper function of nigrostriatal dopaminergic neurons and suggest that Dyrk1A(+/-) mice can be used to study the pathogenesis of motor disorders involving dopaminergic dysfunction.
Asunto(s)
Dopamina/metabolismo , Actividad Motora/fisiología , Neostriado/enzimología , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Tirosina Quinasas/metabolismo , Sustancia Negra/enzimología , Animales , Femenino , Heterocigoto , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microdiálisis , Vías Nerviosas/metabolismo , Tomografía de Emisión de Positrones , Prosencéfalo/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Tirosina Quinasas/genética , Tirosina 3-Monooxigenasa/metabolismo , Quinasas DyrKRESUMEN
The gene encoding the dual-specificity tyrosine-regulated kinase DYRK1A maps to the chromosomal segment HSA21q22.2, which lies within the Down syndrome critical region. The reduction in brain size and behavioral defects observed in mice lacking one copy of the murine homologue Dyrk1A (Dyrk1A+/-) support the idea that this kinase may be involved in monosomy 21 associated mental retardation. However, the structural basis of these behavioral defects remains unclear. In the present work, we have analyzed the microstructure of cortical circuitry in the Dyrk1A+/- mouse and control littermates by intracellular injection of Lucifer Yellow in fixed cortical tissue. We found that labeled pyramidal cells were considerably smaller, less branched and less spinous in the cortex of Dyrk1A+/- mice than in control littermates. These results suggest that Dyrk1A influences the size and complexity of pyramidal cells, and thus their capability to integrate information.
Asunto(s)
Neocórtex/anomalías , Neocórtex/patología , Malformaciones del Sistema Nervioso/genética , Proteínas Serina-Treonina Quinasas/genética , Células Piramidales/patología , Animales , Diferenciación Celular/genética , Forma de la Célula/genética , Espinas Dendríticas/metabolismo , Espinas Dendríticas/patología , Modelos Animales de Enfermedad , Síndrome de Down/genética , Síndrome de Down/patología , Síndrome de Down/fisiopatología , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neocórtex/metabolismo , Malformaciones del Sistema Nervioso/metabolismo , Malformaciones del Sistema Nervioso/patología , Fenotipo , Proteínas Tirosina Quinasas , Células Piramidales/metabolismo , Quinasas DyrKRESUMEN
DYRK1A is a protein kinase proposed to be involved in neurogenesis. Gene-targeting disruption of Dyrk1A in mice leads to decreased body and brain size, with no severe disturbance of behavior. In this study, the authors focused on the motor profile of Dyrk1A(+/-) mice. These mice presented impairment of neuromotor development with decreased activity, suggesting a physiological role of Dyrk1A in the maturation of the neuromotor system. In the adult, a marked hypoactivity and alteration of specific motor parameters were detected. These results are in agreement with the significant expression of Dyrk1A in structures related to motor function and support a role of Dyrk1A in the control of motor function.
Asunto(s)
Haplotipos , Actividad Motora/fisiología , Proteínas Serina-Treonina Quinasas/deficiencia , Proteínas Tirosina Quinasas/deficiencia , Desempeño Psicomotor/fisiología , Factores de Edad , Análisis de Varianza , Animales , Animales Recién Nacidos , Conducta Exploratoria/fisiología , Fuerza de la Mano/fisiología , Ratones , Ratones Endogámicos , Ratones Transgénicos , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/fisiología , Proteínas Tirosina Quinasas/genética , Proteínas Tirosina Quinasas/fisiología , Tiempo de Reacción/fisiología , Caminata/fisiología , Quinasas DyrKRESUMEN
La nutrición artificial domiciliaria es una modalidad de tratamiento nutricional, utilizada cada vez con más frecuencia, que tiene como objeto contribuir a mejorar la calidad de vida de los pacientes que necesitan tratamiento nutricional pero cuya permanencia en el hospital no resulta estrictamente necesaria. En esta revisión se comentan los aspectos relativos a la regulación legal, la prescripción, el control y la educación necesarios en esta modalidad de tratamiento. Tanto la nutrición enteral domiciliaria como la nutrición parenteral domiciliaria precisan un manejo adecuado por parte del paciente y/o sus cuidadores, y los responsables de los equipos de nutrición debemos asegurarnos de que aquéllos lo comprendan y lo realicen de forma adecuada. El papel fundamental del endocrinólogo, como responsable del equipo de soporte nutricional, debe ser coordinar todas las acciones necesarias para el éxito del tratamiento y el control clínico de los pacientes. También se comentan aspectos relativos al futuro de la nutrición artificial domiciliaria (AU)
Asunto(s)
Alimentación con Biberón/métodos , Alimentación con Biberón , Calidad de Vida , Apoyo Nutricional/métodos , Apoyo Nutricional , Tratamiento Domiciliario/métodos , Indicadores de Salud , Indicadores de Calidad de Vida , Legislación/normas , Legislación/organización & administración , Legislación , Nutrición Parenteral/métodos , Nutrición Parenteral , España/epidemiología , Selección de Paciente , Deshidratación/complicaciones , Diarrea/complicaciones , Síndrome de Dificultad Respiratoria del Recién Nacido/complicacionesRESUMEN
Pendred syndrome is an autosomal-recessive disorder characterized by congenital sensorineural hearing loss combined with goiter. This disorder may account for up to 10% of cases of hereditary deafness. The disease gene (PDS/SLC26A4) has been mapped to chromosome 7q22-q31 and encodes a chloride-iodide transport protein. Mutations in this gene are also a cause of non-syndromic autosomal recessive hearing impairment (DFNB4). We have analyzed the PDS/SLC26A4 gene in Spanish and Italian families and we have detected five novel mutations (X781W, T132I, IVS2-2A>G, Y556H and 406del5).
Asunto(s)
Proteínas Portadoras/genética , Pérdida Auditiva Sensorineural/genética , Proteínas de Transporte de Membrana , Alelos , Empalme Alternativo/genética , ADN/química , ADN/genética , Análisis Mutacional de ADN , Salud de la Familia , Genotipo , Bocio/complicaciones , Bocio/genética , Pérdida Auditiva Sensorineural/complicaciones , Humanos , Italia , Región Mediterránea , Mutación , Mutación Missense , Fenotipo , Eliminación de Secuencia , España , Transportadores de Sulfato , SíndromeRESUMEN
The present study was aimed at establishing the differences in the neurodevelopmental profile between two F2 lines derived from two F1 hybrid mouse strains (129 x C57BL/6 and C57BL/6 x SJL). The choice of the given strains was based on the frequent use of these mice in transgenic research. For the neurodevelopment phenotyping, we employed a test battery consisting of 23 somatometric, sensorial and motor tests. Significant variations between the strains were established in different functional domains. Some specific delays in the appearance of developmental landmarks were observed in F2 mice derived from crosses of F1 C57BL/6 x 129, whereas they acquired early developmental functions, such as the righting reflex, sooner than C57BL/6 x SJL-derived mice. C57BL/6 x 129 F2 offspring were spontaneously hypoactive, and their poorer motor performance was confirmed by low performance in the negative geotaxis test. However, there were no differences in the general psychomotor development as shown by the good performance in the homing test in both F2 lines. Both strains were susceptible to the handling procedures used, presenting a similar alteration in the response observed in the homing test as compared to nonhandled control mice. In conclusion, our work highlights the importance of the genetic background for transgenesis experiments and also the need for well-established testing protocols to obtain sufficient information at the first stage of behavioral screening of genetically modified mice.
Asunto(s)
Animales Modificados Genéticamente/genética , Actividad Motora/genética , Destreza Motora/fisiología , Animales , Animales Recién Nacidos , Cruzamiento/métodos , Femenino , Manejo Psicológico , Masculino , Ratones , Ratones Endogámicos C57BL , Embarazo , Especificidad de la EspecieRESUMEN
Pendred syndrome is an autosomal-recessive disorder characterized by congenital sensorineural hearing loss combined with goiter. This disorder may account for up to 10% of cases of hereditary deafness. The disease gene (PDS/SLC26A4) has been mapped to chromosome 7q22-q31 and encodes a chloride-iodide transport protein. Mutations in this gene are also a cause of non-syndromic autosomal recessive hearing impairment (DFNB4). We have analyzed the PDS/SLC26A4 gene in Spanish and Italian families and we have detected five new mutations (X871M, T132I, IVS1-2A>G, Y556H and 406del5).
Asunto(s)
Proteínas Portadoras/genética , Trastornos de la Audición/genética , Proteínas de Transporte de Membrana , Alelos , ADN/química , ADN/genética , Análisis Mutacional de ADN , Salud de la Familia , Genotipo , Bocio/complicaciones , Bocio/genética , Pérdida Auditiva Sensorineural/complicaciones , Pérdida Auditiva Sensorineural/genética , Humanos , Italia , Mutación , Fenotipo , España , Transportadores de Sulfato , SíndromeRESUMEN
The availability of the recently published DNA sequence of human chromosome 21 (HSA21) is a landmark contribution that will have an immediate impact on the study of the role of specific genes to Down syndrome (DS). Trisomy 21 or DS is the only autosomal aneuploidy that is not lethal in the fetal or early postnatal period. DS phenotypes show variable penetrance, affecting many different organs, including brain (mental retardation, early onset of Alzheimer's disease, AD), muscle (hypotonia), skeleton, and blood. DS phenotypes may stem directly from the cumulative effect of overexpression of specific HSA21 gene products or indirectly through the interaction of these gene products with the whole genome, transcriptome, or proteome. Mouse genetic models have played an important role in the elucidation of the contribution of specific genes to the DS phenotype. To date, the strategies used for modeling DS in mice have been three: (1) to assess single-gene contributions to DS phenotype, using transgenic techniques to create models overexpressing single or combinations of genes, (2) to assess the effects of overexpressing large foreign DNA pieces, introduced on yeast artificial chromosomes (YACs) or bacterial artificial chromosomes (BACs) into transgenic mice, and (3) mouse trisomies that carry all or part of MMU16, which has regions of conserved homology with HSA21. Here we review the existing murine models and the relevance of their contribution to DS research.
Asunto(s)
Modelos Animales de Enfermedad , Síndrome de Down/genética , Animales , Conducta Animal/fisiología , Encéfalo/patología , Cromosomas Humanos Par 21 , Expresión Génica/fisiología , Genética Conductual , Humanos , Ratones , Ratones Transgénicos , Modelos GenéticosRESUMEN
Mutations in the unconventional myosin VI gene, Myo6, are associated with deafness and vestibular dysfunction in the Snell's waltzer (sv) mouse. The corresponding human gene, MYO6, is located on chromosome 6q13. We describe the mapping of a new deafness locus, DFNA22, on chromosome 6q13 in a family affected by a nonsyndromic dominant form of deafness (NSAD), and the subsequent identification of a missense mutation in the MYO6 gene in all members of the family with hearing loss.
Asunto(s)
Cromosomas Humanos Par 6 , Sordera/genética , Cadenas Pesadas de Miosina/genética , Secuencia de Aminoácidos , Animales , Mapeo Cromosómico , Modelos Animales de Enfermedad , Humanos , Ratones , Modelos Moleculares , Datos de Secuencia Molecular , Mutación , Cadenas Pesadas de Miosina/química , Linaje , Conformación Proteica , Homología de Secuencia de AminoácidoRESUMEN
Mutations in the connexin 31 (GJB3) gene have been found in subjects with dominant and recessive deafness and in patients with erythrokeratodermia variabilis. We report here a dominant mutation in the GJB3 gene (D66del) in a family affected with peripheral neuropathy and sensorineural hearing impairment. A wide range of disease severity for peripheral neuropathy, from asymptomatic cases to subjects with chronic skin ulcers in their feet and osteomyelitis leading to amputations, was detected in D66del patients. Mild, often asymmetrical, hearing impairment was found in all but one patient with mutation D66del of this family and the same mutation was present in an independent family ascertained because of hearing impairment. We have found mouse connexin 31 (Gjb3) gene expression in the cochlea and in the auditory and sciatic nerves, showing a pattern similar to that of Gjb1 (connexin 32), of which the human ortholog (GJB1) is involved in X-linked peripheral neuropathy. This expression pattern, together with auditory-evoked brainstem anomalous response in D66del patients, indicates that hearing impairment due to GJB3 mutations involves alterations in both the cochlea and the auditory nerve. Peripheral neuropathy is the third phenotypic alteration linked to GJB3 mutations, which enlarges the list of genes that cause this group of heterogeneous disorders.
Asunto(s)
Nervio Coclear/metabolismo , Conexinas/genética , Pérdida Auditiva Sensorineural/genética , Enfermedades del Sistema Nervioso Periférico/genética , Sistema Nervioso Periférico/metabolismo , Adulto , Animales , Conexinas/biosíntesis , Análisis Mutacional de ADN , Femenino , Expresión Génica , Humanos , Hibridación in Situ , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Mutación , Linaje , Nervio Ciático/patologíaRESUMEN
We have demonstrated that DSCR1 acts as a negative regulator of calcineurin-mediated signaling and that its transcript is overexpressed in the Down syndrome (DS) fetal brain. To evaluate the possible involvement of DSCR1 in DS, we have cloned the mouse gene and analyzed its expression pattern in the central nervous system (CNS). Early expression of Dscr1 is detected mainly in the heart tube and in the CNS in rhombomere 4 and the pretectum. From embryonic day 14.5 onwards, Dscr1 is widely distributed in the CNS but becomes more restricted as the brain matures. We confirmed its neuronal expression pattern in the adult, preferentially in Purkinje and pyramidal cells, by double labeling with glial fibrillary acidic protein. We also show that although Dscr1 is present in trisomy in the Ts65Dn mouse, the adult brain expression pattern is not significantly altered. This expression pattern indicated that Dscr1 is a developmentally regulated gene involved in neurogenesis and cardiogenesis and suggests that it may contribute to the alterations observed in these organ systems in DS patients.
Asunto(s)
Calcineurina/metabolismo , Sistema Nervioso Central/embriología , Regulación del Desarrollo de la Expresión Génica , Corazón/embriología , Proteínas Musculares/fisiología , Neuronas/metabolismo , Animales , Northern Blotting , Encéfalo/embriología , Clonación Molecular , ADN Complementario/metabolismo , Proteínas de Unión al ADN , Biblioteca de Genes , Humanos , Hibridación in Situ , Hibridación Fluorescente in Situ , Péptidos y Proteínas de Señalización Intracelular , Ratones , Transducción de Señal , Factores de Tiempo , Distribución Tisular , Trisomía/genéticaRESUMEN
The availability of the DNA sequence of human chromosome 21 (HSA21) is a landmark contribution that will have an immediate impact on the study of the role of specific genes to Down syndrome (DS). Trisomy 21, full or partial, is a major cause of mental retardation and other phenotypic abnormalities, collectively known as Down syndrome (DS), a disorder affecting 1 in 700 births. The identification of genes on HSA21 and the elucidation of the function of the proteins encoded by these genes have been a major challenge for the human genome project and for research in DS. Over 100 of the estimated 300-500 genes of HSA21 have been identified, but the function of most remains largely unknown. It is believed that the overexpression of an unknown number of HSA21 genes is directly or indirectly responsible for the mental retardation and the other clinical features of DS. For this reason, HSA21 genes that are expressed in tissues affected in DS patients are of special interest.
Asunto(s)
Proteínas Adaptadoras del Transporte Vesicular , Síndrome de Down/genética , Genoma Humano , Secretasas de la Proteína Precursora del Amiloide , Ácido Aspártico Endopeptidasas/genética , Proteínas Portadoras/genética , Proteínas de Unión al ADN , Endopeptidasas , Genómica , Humanos , Péptidos y Proteínas de Señalización Intracelular , Proteínas Musculares/genética , Proteínas Serina-Treonina Quinasas/genética , Proteínas Tirosina Quinasas/genética , Quinasas DyrKRESUMEN
Mutations in the GJB2 gene have been identified in many patients with childhood deafness, 35delG being the most common mutation in Caucasoid populations. We have analyzed a total of 576 families/unrelated patients with recessive or sporadic deafness from Italy and Spain, 193 of them being referred as autosomal recessive, and the other 383 as apparently sporadic cases (singletons). Of the 1,152 unrelated GJB2 chromosomes analyzed from these patients, 37% had GJB2 mutations. Twenty-three different mutations were detected (1 in-frame deletion, 4 nonsense, 5 frameshift, and 13 missense mutations). Mutation 35delG was the most common, accounting for 82% of all GJB2 deafness alleles. The relative frequency of 35delG in Italy and Spain was different, representing 88% of the alleles in Italian patients and only 55% in the Spanish cases. Eight non-35delG mutations were detected more than once (V37I, E47X, 167delT, L90P, 312de114, 334delAA, R143W, and R184P), with relative frequencies ranging between 0.5 and 1.6% of the GJB2 deafness alleles. The information based on conservation of amino acid residues, coexistence with a second GJB2 mutation or absence of the mutation in non-deaf control subjects, suggests that most of these missense changes should be responsible for the deafness phenotype.
Asunto(s)
Conexinas/genética , Pérdida Auditiva Sensorineural/genética , Mutación , Alelos , Niño , Conexina 26 , Mutación del Sistema de Lectura , Eliminación de Gen , Genes Recesivos , Pérdida Auditiva Sensorineural/congénito , Humanos , Mutación MissenseRESUMEN
Connexin31 (GJB3) has been associated with hearing impairment and erythrokeratodermia variabilis. We have analyzed this gene in samples from patients with peripheral neuropathies, deafness and controls and have found several single nucleotide polymorphisms (SNPs). In the noncoding exon 1 of GJB3 two small deletions, 581del2 and 632del4 (GenBank accession number AF052692), were found at frequencies of 30% and 14%, respectively. In exon 2 we found two amino acid changes, R32W (1227C-T) and V200I (1731G-A), and three nucleotide variants not affecting the amino acid sequence, 1610G-A, 1700C-T and 1931C-T. Most of these changes were found at similar frequencies in patients with deafness, patients with peripheral neuropathies and control subjects. V200I, 1700C-T and 1610G-A were found associated in three unrelated patients with deafness and in a fourth patient with peripheral neuropathy, but were not detected in control subjects.