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1.
FASEB J ; 34(3): 4573-4590, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-32020675

RESUMEN

Aberrant expression of the transcription factor double homeobox protein 4 (DUX4) can lead to a number of diseases including facio-scapulo-humeral muscular dystrophy (FSHD), acute lymphoblastic leukemia, and sarcomas. Inhibition of DUX4 may represent a therapeutic strategy for these diseases. By applying Systematic Evolution of Ligands by EXponential Enrichment (SELEX), we identified aptamers against DUX4 with specific secondary structural elements conveying high affinity to DUX4 as assessed by fluorescence resonance energy transfer and fluorescence polarization techniques. Sequences analysis of these aptamers revealed the presence of two consensus DUX4 motifs in a reverse complementary fashion forming hairpins interspersed with bulge loops at distinct positions that enlarged the binding surface with the DUX4 protein, as determined by crystal structure analysis. We demonstrate that insertion of specific structural elements into transcription factor binding oligonucleotides can enhance specificity and affinity.


Asunto(s)
Aptámeros de Nucleótidos/química , Proteínas de Homeodominio/metabolismo , Distrofia Muscular Facioescapulohumeral/metabolismo , Técnica SELEX de Producción de Aptámeros/métodos , Proteínas de Homeodominio/química , Proteínas de Homeodominio/genética , Humanos , Masculino , Modelos Moleculares , Factor de Transcripción PAX7/química , Factor de Transcripción PAX7/genética , Factor de Transcripción PAX7/metabolismo
2.
J Biol Chem ; 291(33): 17165-77, 2016 08 12.
Artículo en Inglés | MEDLINE | ID: mdl-27298317

RESUMEN

Myotonic dystrophy type I (DM1) is a disabling neuromuscular disease with no causal treatment available. This disease is caused by expanded CTG trinucleotide repeats in the 3' UTR of the dystrophia myotonica protein kinase gene. On the RNA level, expanded (CUG)n repeats form hairpin structures that sequester splicing factors such as muscleblind-like 1 (MBNL1). Lack of available MBNL1 leads to misregulated alternative splicing of many target pre-mRNAs, leading to the multisystemic symptoms in DM1. Many studies aiming to identify small molecules that target the (CUG)n-MBNL1 complex focused on synthetic molecules. In an effort to identify new small molecules that liberate sequestered MBNL1 from (CUG)n RNA, we focused specifically on small molecules of natural origin. Natural products remain an important source for drugs and play a significant role in providing novel leads and pharmacophores for medicinal chemistry. In a new DM1 mechanism-based biochemical assay, we screened a collection of isolated natural compounds and a library of over 2100 extracts from plants and fungal strains. HPLC-based activity profiling in combination with spectroscopic methods were used to identify the active principles in the extracts. The bioactivity of the identified compounds was investigated in a human cell model and in a mouse model of DM1. We identified several alkaloids, including the ß-carboline harmine and the isoquinoline berberine, that ameliorated certain aspects of the DM1 pathology in these models. Alkaloids as a compound class may have potential for drug discovery in other RNA-mediated diseases.


Asunto(s)
Regiones no Traducidas 3' , Alcaloides/farmacología , Proteínas de Unión al ADN , Modelos Biológicos , Distrofia Miotónica/tratamiento farmacológico , Proteínas de Unión al ARN , Expansión de Repetición de Trinucleótido , Alcaloides/química , Alcaloides/aislamiento & purificación , Empalme Alternativo/efectos de los fármacos , Animales , Línea Celular , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Evaluación Preclínica de Medicamentos/métodos , Humanos , Ratones , Distrofia Miotónica/genética , Distrofia Miotónica/metabolismo , Distrofia Miotónica/patología , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/metabolismo
3.
Skelet Muscle ; 14(1): 11, 2024 May 21.
Artículo en Inglés | MEDLINE | ID: mdl-38769542

RESUMEN

BACKGROUND: Myotonic Dystrophy type I (DM1) is the most common muscular dystrophy in adults. Previous reports have highlighted that neuromuscular junctions (NMJs) deteriorate in skeletal muscle from DM1 patients and mouse models thereof. However, the underlying pathomechanisms and their contribution to muscle dysfunction remain unknown. METHODS: We compared changes in NMJs and activity-dependent signalling pathways in HSALR and Mbnl1ΔE3/ΔE3 mice, two established mouse models of DM1. RESULTS: Muscle from DM1 mouse models showed major deregulation of calcium/calmodulin-dependent protein kinases II (CaMKIIs), which are key activity sensors regulating synaptic gene expression and acetylcholine receptor (AChR) recycling at the NMJ. Both mouse models exhibited increased fragmentation of the endplate, which preceded muscle degeneration. Endplate fragmentation was not accompanied by changes in AChR turnover at the NMJ. However, the expression of synaptic genes was up-regulated in mutant innervated muscle, together with an abnormal accumulation of histone deacetylase 4 (HDAC4), a known target of CaMKII. Interestingly, denervation-induced increase in synaptic gene expression and AChR turnover was hampered in DM1 muscle. Importantly, CaMKIIß/ßM overexpression normalized endplate fragmentation and synaptic gene expression in innervated Mbnl1ΔE3/ΔE3 muscle, but it did not restore denervation-induced synaptic gene up-regulation. CONCLUSIONS: Our results indicate that CaMKIIß-dependent and -independent mechanisms perturb synaptic gene regulation and muscle response to denervation in DM1 mouse models. Changes in these signalling pathways may contribute to NMJ destabilization and muscle dysfunction in DM1 patients.


Asunto(s)
Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina , Modelos Animales de Enfermedad , Músculo Esquelético , Distrofia Miotónica , Unión Neuromuscular , Distrofia Miotónica/genética , Distrofia Miotónica/metabolismo , Distrofia Miotónica/fisiopatología , Animales , Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/metabolismo , Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/genética , Unión Neuromuscular/metabolismo , Músculo Esquelético/metabolismo , Músculo Esquelético/inervación , Músculo Esquelético/patología , Ratones , Humanos , Histona Desacetilasas/metabolismo , Histona Desacetilasas/genética , Receptores Colinérgicos/metabolismo , Receptores Colinérgicos/genética , Masculino , Ratones Endogámicos C57BL
5.
J Biol Chem ; 287(13): 10344-10354, 2012 Mar 23.
Artículo en Inglés | MEDLINE | ID: mdl-22318734

RESUMEN

Dysferlin is a transmembrane protein implicated in surface membrane repair of muscle cells. Mutations in dysferlin cause the progressive muscular dystrophies Miyoshi myopathy, limb girdle muscular dystrophy 2B, and distal anterior compartment myopathy. Dysferlinopathies are inherited in an autosomal recessive manner, and many patients with this disease harbor mis-sense mutations in at least one of their two pathogenic DYSF alleles. These patients have significantly reduced or absent dysferlin levels in skeletal muscle, suggesting that dysferlin encoded by mis-sense alleles is rapidly degraded by the cellular quality control system. We reasoned that mis-sense mutated dysferlin, if salvaged from degradation, might be biologically functional. We used a dysferlin-deficient human myoblast culture harboring the common R555W mis-sense allele and a DYSF-null allele, as well as control human myoblast cultures harboring either two wild-type or two null alleles. We measured dysferlin protein and mRNA levels, resealing kinetics of laser-induced plasmalemmal wounds, myotube formation, and cellular viability after treatment of the human myoblast cultures with the proteasome inhibitors lactacystin or bortezomib (Velcade). We show that endogenous R555W mis-sense mutated dysferlin is degraded by the proteasomal system. Inhibition of the proteasome by lactacystin or Velcade increases the levels of R555W mis-sense mutated dysferlin. This salvaged protein is functional as it restores plasma membrane resealing in patient-derived myoblasts and reverses their deficit in myotube formation. Bortezomib and lactacystin did not cause cellular toxicity at the regimen used. Our results raise the possibility that inhibition of the degradation pathway of mis-sense mutated dysferlin could be used as a therapeutic strategy for patients harboring certain dysferlin mis-sense mutations.


Asunto(s)
Acetilcisteína/análogos & derivados , Antineoplásicos/farmacología , Ácidos Borónicos/farmacología , Inhibidores de Cisteína Proteinasa/farmacología , Miopatías Distales/tratamiento farmacológico , Proteínas de la Membrana/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Proteínas Musculares/metabolismo , Atrofia Muscular/tratamiento farmacológico , Distrofia Muscular de Cinturas/tratamiento farmacológico , Mutación Missense , Inhibidores de Proteasoma , Proteolisis/efectos de los fármacos , Pirazinas/farmacología , Acetilcisteína/farmacología , Alelos , Sustitución de Aminoácidos , Bortezomib , Células Cultivadas , Miopatías Distales/genética , Miopatías Distales/metabolismo , Miopatías Distales/patología , Disferlina , Humanos , Proteínas de la Membrana/genética , Fibras Musculares Esqueléticas/patología , Proteínas Musculares/genética , Atrofia Muscular/genética , Atrofia Muscular/metabolismo , Atrofia Muscular/patología , Distrofia Muscular de Cinturas/genética , Distrofia Muscular de Cinturas/metabolismo , Distrofia Muscular de Cinturas/patología , Complejo de la Endopetidasa Proteasomal/genética , Complejo de la Endopetidasa Proteasomal/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo
6.
Neurobiol Dis ; 49: 221-31, 2013 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22940629

RESUMEN

Charcot-Marie-Tooth disease type 1A (CMT1A) is a hereditary demyelinating peripheral neuropathy caused by the duplication of the PMP22 gene. Demyelination precedes the occurrence of clinical symptoms that correlate with axonal degeneration. It was postulated that a disturbed axon-glia interface contributes to altered myelination consequently leading to axonal degeneration. In this study, we examined the expression of MAG and Necl4, two critical adhesion molecules that are present at the axon-glia interface, in sural nerve biopsies of CMT1A patients and in peripheral nerves of mice overexpressing human PMP22, an animal model for CMT1A. We show an increase in the expression of MAG and a strong decrease of Necl4 in biopsies of CMT1A patients as well as in CMT1A mice. Expression analysis revealed that MAG is strongly upregulated during peripheral nerve maturation, whereas Necl4 expression remains very low. Ablating MAG in CMT1A mice results in separation of axons from their myelin sheath. Our data show that MAG is important for axon-glia contact in a model for CMT1A, and suggest that its increased expression in CMT1A disease has a compensatory role in the pathology of the disease. Thus, we demonstrate that MAG together with other adhesion molecules such as Necl4 is important in sustaining axonal integrity.


Asunto(s)
Axones/metabolismo , Enfermedad de Charcot-Marie-Tooth/metabolismo , Vaina de Mielina/metabolismo , Glicoproteína Asociada a Mielina/metabolismo , Nervio Sural/metabolismo , Adulto , Anciano de 80 o más Años , Animales , Axones/patología , Moléculas de Adhesión Celular/metabolismo , Enfermedad de Charcot-Marie-Tooth/patología , Modelos Animales de Enfermedad , Femenino , Humanos , Inmunoglobulinas/metabolismo , Masculino , Ratones Noqueados , Ratones Transgénicos , Persona de Mediana Edad , Proteínas de la Mielina/genética , Proteínas de la Mielina/metabolismo , Vaina de Mielina/patología , Glicoproteína Asociada a Mielina/genética , Nervio Sural/patología , Adulto Joven
7.
J Peripher Nerv Syst ; 16 Suppl 1: 30-3, 2011 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-21696494

RESUMEN

DNA microarray analysis is a powerful tool for simultaneous analysis and comparison of gene products expressed in normal and diseased tissues. We used this technique to identify differentially expressed genes (DEGs) in nerve biopsy samples of chronic inflammatory demyelinating polyneuropathy (CIDP) and vasculitic neuropathy (VAS) patients. We found novel previously uncharacterized genes of relevance to CIDP or VAS pathogenesis. Of particular interest in CIDP were tachykinin precursor 1, which may be involved in pain mediation, stearoyl-co-enzyme A (CoA) desaturase, which may be a marker for remyelination, HLA-DQB1, CD69, an early T-cell activation gene, MSR1, a macrophage scavenger receptor, and PDZ and LIM domain 5 (PDLIM5), a factor regulating nuclear factor (NF)-kappa B activity. Genes upregulated in VAS included IGLJ3, IGHG3, IGKC, and IGL, which all function in B-cell selection or antigen recognition of B cells. Other upregulated genes included chemokines, such as CXCL9 and CCR2, as well as CPA3, a mast cell carboxypeptidase. Allograft inflammatory factor-1 (AIF-1), a modulator of immune response was upregulated both in CIDP and VAS. Microarray-based analysis of human sural nerve biopsies showed distinct gene expression patterns in CIDP and VAS. DEGs might provide clues to the pathogenesis of the diseases and be potential targets for therapeutics.


Asunto(s)
Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/genética , Vasculitis/genética , Biomarcadores/análisis , Biopsia , Expresión Génica , Perfilación de la Expresión Génica , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Enfermedades del Sistema Nervioso Periférico/genética , Enfermedades del Sistema Nervioso Periférico/inmunología , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/inmunología , Nervio Sural , Vasculitis/inmunología
8.
Hepatology ; 50(3): 871-9, 2009 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-19551907

RESUMEN

UNLABELLED: Intrahepatic bile duct (IHBD) development begins with the differentiation of hepatoblasts into a single continuous biliary epithelial cell (BEC) layer, called the ductal plate. During ductal plate remodeling, tubular structures arise at distinct sites of the ductal plate, forming bile ducts that dilate into the biliary tree. Alagille syndrome patients, who suffer from bile duct paucity, carry Jagged1 and Notch2 mutations, indicating that Notch2 signaling is important for IHBD development. To clarify the role of Notch2 in BEC differentiation, tubulogenesis, and BEC survival, we developed a mouse model for conditional expression of activated Notch2 in the liver. We show that expression of the intracellular domain of Notch2 (Notch2ICD) differentiates hepatoblasts into BECs, which form additional bile ducts in periportal regions and ectopic ducts in lobular regions. Additional ducts in periportal regions are maintained into adulthood and connect to the biliary tight junction network, resulting in an increased number of bile ducts per portal tract. Remarkably, Notch2ICD-expressing ductal plate remnants were not eliminated during postnatal development, implicating Notch2 signaling in BEC survival. Ectopic ducts in lobular regions did not persist into adulthood, indicating that local signals in the portal environment are important for maintaining bile ducts. CONCLUSION: Notch2 signaling regulates BEC differentiation, the induction of tubulogenesis during IHBD development, and BEC survival.


Asunto(s)
Conductos Biliares Intrahepáticos/embriología , Células Epiteliales/fisiología , Receptor Notch2/fisiología , Animales , Diferenciación Celular/fisiología , Ratones , Ratones Transgénicos
9.
Mol Biol Cell ; 17(8): 3651-63, 2006 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-16760430

RESUMEN

We identified a direct interaction between the neuronal transmembrane protein calsyntenin-1 and the light chain of Kinesin-1 (KLC1). GST pulldowns demonstrated that two highly conserved segments in the cytoplasmic domain of calsyntenin-1 mediate binding to the tetratricopeptide repeats of KLC1. A complex containing calsyntenin-1 and the Kinesin-1 motor was isolated from developing mouse brain and immunoelectron microscopy located calsyntenin-1 in association with tubulovesicular organelles in axonal fiber tracts. In primary neuronal cultures, calsyntenin-1-containing organelles were aligned along microtubules and partially colocalized with Kinesin-1. Using live imaging, we showed that these organelles are transported along axons with a velocity and processivity typical for fast axonal transport. Point mutations in the two kinesin-binding segments of calsyntenin-1 significantly reduced binding to KLC1 in vitro, and vesicles bearing mutated calsyntenin-1 exhibited a markedly altered anterograde axonal transport. In summary, our results indicate that calsyntenin-1 links a certain type of vesicular and tubulovesicular organelles to the Kinesin-1 motor.


Asunto(s)
Proteínas de Unión al Calcio/metabolismo , Vesículas Citoplasmáticas/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismo , Secuencia de Aminoácidos , Animales , Proteínas de Unión al Calcio/química , Secuencia Conservada , Conos de Crecimiento/metabolismo , Células HeLa , Humanos , Cinesinas , Ratones , Datos de Secuencia Molecular , Mutación/genética , Unión Proteica , Transporte de Proteínas , Ratas , Ratas Sprague-Dawley
10.
Nucleic Acids Res ; 30(10): 2202-11, 2002 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-12000840

RESUMEN

Neuroserpin is an axonally secreted serine protease inhibitor expressed in the nervous system that protects neurons from ischemia-induced apoptosis. Mutant neuroserpin forms have been found polymerized in inclusion bodies in a familial autosomal encephalopathy causing dementia, or associated with epilepsy. Regulation of neuroserpin expression is mostly unknown. Here we demonstrate that neuroserpin mRNA and the RNA-binding protein HuD are co-expressed in the rat central nervous system, and that HuD binds neuroserpin mRNA in vitro with high affinity. Gel-shift, supershift and T1 RNase assays revealed three HuD-binding sequences in the 3'-untranslated region (3'-UTR) of neuroserpin mRNA. They are AU-rich and 20, 51 and 19 nt in length. HuD binding to neuroserpin mRNA was also demonstrated in extracts of PC12 pheochromocytoma cells. Additionally, ectopic expression of increasing amounts of HuD in these cells results in the accumulation of neuroserpin 3'-UTR mRNA. Furthermore, stably transfected PC12 cells over-expressing HuD contain increased levels of both neuroserpin mRNAs (3.0 and 1.6 kb) and protein. Our results indicate that HuD stabilizes neuroserpin mRNA by binding to specific AU-rich sequences in its 3'-UTR, which prolongs the mRNA lifetime and increases protein level.


Asunto(s)
Regiones no Traducidas 3'/genética , Proteínas del Tejido Nervioso/metabolismo , Neuropéptidos/metabolismo , Regiones Promotoras Genéticas/genética , ARN Mensajero/metabolismo , Proteínas de Unión al ARN/metabolismo , Serpinas/metabolismo , Animales , Sitios de Unión , Unión Competitiva , Encéfalo/metabolismo , Proteínas ELAV , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Hibridación in Situ , Proteínas del Tejido Nervioso/genética , Neuropéptidos/genética , Células PC12 , Unión Proteica , Estabilidad del ARN , ARN Mensajero/genética , Proteínas de Unión al ARN/genética , Ratas , Ratas Wistar , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Serpinas/genética , Transcripción Genética , Neuroserpina
11.
Skelet Muscle ; 5: 32, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26464793

RESUMEN

BACKGROUND: Mouse models of dysferlinopathies are valuable tools with which to investigate the pathomechanisms underlying these diseases and to test novel therapeutic strategies. One such mouse model is the Dysf (tm1Kcam) strain, which was generated using a targeting vector to replace a 12-kb region of the dysferlin gene and which features a progressive muscular dystrophy. A prerequisite for successful animal studies using genetic mouse models is an accurate genotyping protocol. Unfortunately, the lack of robustness of currently available genotyping protocols for the Dysf (tm1Kcam) mouse has prevented efficient colony management. Initial attempts to improve the genotyping protocol based on the published genomic structure failed. These difficulties led us to analyze the targeted locus of the dysferlin gene of the Dysf (tm1Kcam) mouse in greater detail. METHODS: In this study we resequenced and analyzed the targeted locus of the Dysf (tm1Kcam) mouse and developed a novel PCR protocol for genotyping. RESULTS: We found that instead of a deletion, the dysferlin locus in the Dysf (tm1Kcam) mouse carries a targeted insertion. This genetic characterization enabled us to establish a reliable method for genotyping of the Dysf (tm1Kcam) mouse, and thus has made efficient colony management possible. CONCLUSION: Our work will make the Dysf (tm1Kcam) mouse model more attractive for animal studies of dysferlinopathies.

12.
Swiss Med Wkly ; 144: w13916, 2014 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-24554202

RESUMEN

Muscular dystrophies are classically subdivided according to their clinical phenotype, and were historically defined as progressive myopathies in which muscle biopsies demonstrate muscle fibre necrosis and regeneration, as well as replacement of muscle fibres by adipose and connective tissue. In recent years, great progress has been made in identifying the genetic basis of many myopathies, thereby presenting opportunities to develop therapeutic strategies that act on specific molecular pathomechanisms. The different therapeutic strategies and their molecular targets will be reviewed.


Asunto(s)
Distrofina/genética , Terapia Genética , Terapia Molecular Dirigida , Distrofias Musculares/terapia , Animales , Humanos , Distrofias Musculares/genética , Distrofia Muscular de Cinturas/genética , Distrofia Muscular Facioescapulohumeral/genética , Mioblastos/trasplante , Distrofia Miotónica/genética
13.
Neuromuscul Disord ; 23(2): 149-54, 2013 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23140793

RESUMEN

Inclusion body myopathy associated with Paget's disease of the bone and frontotemporal dementia is a rare but highly penetrant autosomal dominant progressive disorder linked to mutations in valosin containing protein (VCP). Here, we characterize a novel mutation in the linker 1 domain of VCP leading to inclusion body myopathy and/or frontotemporal dementia in 3 generations of a Swiss family. A detailed history of several years of clinical follow-up and electrophysiological, radiological and pathological findings are presented. Five out of 6 individuals suffered from progressive myopathy and 2 out of 6 from frontotemporal dementia, respectively. A radiologically suspected Paget's disease of the bone could not been confirmed at autopsy. This case study illustrates that only a subset of individuals shows the full triad of the disease complex and that clinicopathological findings are - when interpreted apart from familial history - hard to distinguish from sporadic inclusion body myositis.


Asunto(s)
Adenosina Trifosfatasas/genética , Proteínas de Ciclo Celular/genética , Demencia Frontotemporal/genética , Distrofia Muscular de Cinturas/genética , Mutación/genética , Miositis por Cuerpos de Inclusión/genética , Osteítis Deformante/genética , Biopsia , Femenino , Demencia Frontotemporal/etnología , Demencia Frontotemporal/patología , Humanos , Masculino , Persona de Mediana Edad , Músculo Esquelético/patología , Distrofia Muscular de Cinturas/etnología , Distrofia Muscular de Cinturas/patología , Miositis por Cuerpos de Inclusión/etnología , Miositis por Cuerpos de Inclusión/patología , Osteítis Deformante/etnología , Osteítis Deformante/patología , Linaje , Suiza , Proteína que Contiene Valosina
14.
J Neuroimmunol ; 225(1-2): 184-9, 2010 Aug 25.
Artículo en Inglés | MEDLINE | ID: mdl-20542337

RESUMEN

To investigate molecular mechanisms of peripheral nerve vasculitis, gene expression patterns in archived frozen sural nerve biopsies from patients with vasculitic neuropathy were compared to control nerves by DNA microarray technology. There was a striking upregulation of mRNA of genes involved in immune system processes. Of special interest was the activation of immunoglobulin genes, such as IGLJ3, IGHG3, IGKC, and IGL, and of several chemokines, such as CXCL9 or CCR2. Genes involved in vascular proliferation or remodelling such as CXC31 and AIF were also upregulated. Among the downregulated genes were the Krüppel-Like Transcription Factors KLF2, KLF4 and the nuclear orphan receptor NR4A1 genes known to be involved in endothelial cell activation. Thus, this gene expression profile analysis revealed that in peripheral nerve vasculitis a prominent activation of immune response related genes as well as genes involved in vascular proliferation is taken place, while genes inhibiting endothelial cell activation are down regulated. These data point to interesting mechanistic clues to the molecular pathogenesis of vasculitic neuropathies.


Asunto(s)
Perfilación de la Expresión Génica/métodos , Regulación de la Expresión Génica/fisiología , Enfermedades del Sistema Nervioso Periférico/patología , Nervio Sural/metabolismo , Vasculitis/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biopsia/métodos , Preescolar , Femenino , Humanos , Factor 4 Similar a Kruppel , Factores de Transcripción de Tipo Kruppel/genética , Factores de Transcripción de Tipo Kruppel/metabolismo , Masculino , Miembro 1 del Grupo A de la Subfamilia 4 de Receptores Nucleares/genética , Miembro 1 del Grupo A de la Subfamilia 4 de Receptores Nucleares/metabolismo , Enfermedades del Sistema Nervioso Periférico/complicaciones , Enfermedades del Sistema Nervioso Periférico/genética , Enfermedades del Sistema Nervioso Periférico/metabolismo , Nervio Sural/inmunología , Vasculitis/complicaciones , Vasculitis/genética , Vasculitis/metabolismo
15.
Am J Pathol ; 170(4): 1305-13, 2007 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-17392169

RESUMEN

Intracellular protein deposition due to aggregation caused by conformational alteration is the hallmark of a number of neurodegenerative disorders, including Parkinson's disease, tauopathies, Huntington's disease, and familial encephalopathy with neuroserpin inclusion bodies. The latter is an autosomal dominant disorder caused by point mutations in neuroserpin resulting in its destabilization. Mutant neuroserpin polymerizes and forms intracellular aggregates that eventually lead to neurodegeneration. We generated genetically modified mice expressing the late-onset S49P-Syracuse or the early-onset S52R-Portland mutation of neuroserpin in central nervous system neurons. Mice exhibited morphological, biochemical, and clinical features resembling those found in the human disease. Analysis of brains revealed large intraneuronal inclusions composed exclusively of mutant neuroserpin, accumulating long before the development of clinical symptoms in a time-dependent manner. Clinical symptoms and amount of neuroserpin inclusions correlated with the predicted instability of the protein. The presence of inclusion bodies in subclinical mice indicates that in humans the prevalence of the disease could be higher than anticipated. In addition to shedding light on the pathophysiology of the human disorder, these mice provide an excellent model to study mechanisms of neurodegeneration or establish novel therapies for familial encephalopathy with neuroserpin inclusion bodies and other neurodegenerative diseases with intracellular protein deposition.


Asunto(s)
Encefalopatías/patología , Cuerpos de Inclusión/patología , Proteínas Mutantes/metabolismo , Neuropéptidos/metabolismo , Serpinas/metabolismo , Animales , Western Blotting , Encefalopatías/genética , Encefalopatías/metabolismo , Retículo Endoplásmico Rugoso/metabolismo , Retículo Endoplásmico Rugoso/ultraestructura , Femenino , Humanos , Cuerpos de Inclusión/metabolismo , Cuerpos de Inclusión/ultraestructura , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Ratones Transgénicos , Microscopía Electrónica , Mutación , Neuronas/metabolismo , Neuronas/patología , Neuronas/ultraestructura , Neuropéptidos/genética , Serpinas/genética , Neuroserpina
16.
Mol Cell Neurosci ; 23(3): 473-94, 2003 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-12837630

RESUMEN

Neuroserpin is a neural serpin that inhibits the extracellular protease tissue-type plasminogen activator (tPA). We have generated neuroserpin-deficient mice which are viable and healthy. Zymographic analysis of neuroserpin-deficient brain showed unaltered tPA activity, suggesting that other inhibitors contribute to the regulation of tPA and may compensate for the defect. Analysis of explorative behavior revealed selective reduction of locomotor activity in novel environments, an anxiety-like response on the O-maze, and a neophobic response to novel objects. Mice overexpressing neuroserpin under the control of the Thy1.2 promoter are known to have a reduced brain tPA activity. They showed reduced center exploration in the open-field test and, like neuroserpin-deficient mice, a neophobic phenotype in the novel object test. Our results implicate neuroserpin in the regulation of emotional behavior through a mechanism that is at least in part independent of tPA activity. They are the first evidence for a role of protease inhibitors in mood regulation.


Asunto(s)
Conducta Exploratoria/fisiología , Neuropéptidos/genética , Trastornos Fóbicos/fisiopatología , Serpinas/genética , Afecto/fisiología , Animales , Reacción de Prevención/fisiología , Conducta Animal/fisiología , Espacio Extracelular/enzimología , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes , Actividad Motora/fisiología , Serina Endopeptidasas/metabolismo , Activador de Tejido Plasminógeno/metabolismo , Neuroserpina
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