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1.
Mol Brain ; 13(1): 148, 2020 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-33172478

RESUMO

Neuronal activity-regulated gene transcription underlies plasticity-dependent changes in the molecular composition and structure of neurons. A large number of genes regulated by different neuronal plasticity inducing pathways have been identified, but altered gene expression levels represent only part of the complexity of the activity-regulated transcriptional program. Alternative splicing, the differential inclusion and exclusion of exonic sequence in mRNA, is an additional mechanism that is thought to define the activity-dependent transcriptome. Here, we present a genome wide microarray-based survey to identify exons with increased expression levels at 1, 4 or 8 h following neuronal activity in the murine hippocampus provoked by generalized seizures. We used two different bioinformatics approaches to identify alternative activity-induced exon usage and to predict alternative splicing, ANOSVA (ANalysis Of Splicing VAriation) which we here adjusted to accommodate data from different time points and FIRMA (Finding Isoforms using Robust Multichip Analysis). RNA sequencing, in situ hybridization and reverse transcription PCR validate selected activity-dependent splicing events of previously described and so far undescribed activity-regulated transcripts, including Homer1a, Homer1d, Ania3, Errfi1, Inhba, Dclk1, Rcan1, Cda, Tpm1 and Krt75. Taken together, our survey significantly adds to the comprehensive understanding of the complex activity-dependent neuronal transcriptomic signature. In addition, we provide data sets that will serve as rich resources for future comparative expression analyses.


Assuntos
Processamento Alternativo/genética , Éxons/genética , Neurônios/metabolismo , Animais , Masculino , Camundongos Endogâmicos C57BL , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Reprodutibilidade dos Testes
2.
Int J Biol Macromol ; 153: 625-632, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32165201

RESUMO

This study aims to develop and characterize colorimetric indicator films based on chitosan, polyvinyl alcohol and anthocyanins from jambolan fruit (Syzygium cumini) prepared by casting method. The effect of anthocyanin extract on thickness, microstructure, moisture content, solubility in water, hydrophobicity, chemical structure, color and opacity of films was analyzed. In addition, anthocyanins photodegradation in films as well its application to monitoring shrimp freshness was studied. Significant effect (p < 0.05) of anthocyanin extract from jambolan fruit on the thickness and optical properties of the films was observed. Anthocyanin extract from jambolan fruit was efficiently incorporated and dispersed into film. The films containing anthocyanins showed visible changes from red color to blue color when used to monitor shrimp freshness at several temperatures (between -20 °C and 20 °C). This research reports for the first time information regarding the valorization and application of anthocyanins from jambolan fruit as an alternative for food packaging sector.


Assuntos
Antocianinas/química , Quitosana/química , Conservação de Alimentos , Frutas/química , Álcool de Polivinil/química , Alimentos Marinhos , Syzygium/química
3.
Biol Chem ; 400(9): 1181-1189, 2019 08 27.
Artigo em Inglês | MEDLINE | ID: mdl-31095505

RESUMO

Accumulation of ß-amyloid peptide (Aß) is regarded as a primary cause of Alzheimer's disease (AD). Aß is derived by sequential cleavage of the amyloid precursor protein (APP). Alterations in the subcellular targeting of APP are thought to affect the degree of Aß production. Sorting receptors, such as SorLA, convey subcellular targeting of APP. Dysfunction of SorLA, and likely of the related receptors SorCS1 and SorCS3, cause AD. Nevertheless, disease progression could also provoke altered expression of the receptors. Here, we assessed if Aß plaque formation promotes altered expression of SorLA, SorCS1 and SorCS3. We analyzed transcript levels during aging and after amyloidosis in brain areas characterized by early amyloid plaque formation in an AD mouse model (APPPS1) and wild types. We observed stable expression levels during aging (1-12 months). After plaque formation, SorCS1 and SorLA expression were markedly reduced in the frontal cerebral cortex and to a minor extent in the hippocampus, whereas SorCS3 expression was solely reduced in the frontal cerebral cortex. Our results indicate that disease progression, associated with Aß accumulation, can negatively regulate expression of the receptors.


Assuntos
Amiloidose/genética , Regulação para Baixo , Proteínas de Membrana Transportadoras/genética , Proteínas do Tecido Nervoso/genética , Receptores de Superfície Celular/genética , Receptores de LDL/genética , Doença de Alzheimer/metabolismo , Amiloidose/metabolismo , Animais , Encéfalo/metabolismo , Camundongos , Camundongos Transgênicos , RNA Mensageiro/genética
4.
Sci Rep ; 7: 45101, 2017 03 28.
Artigo em Inglês | MEDLINE | ID: mdl-28349920

RESUMO

Activity-dependent alteration of the transcriptional program is central for shaping neuronal connectivity. Constitutively expressed transcription factors orchestrate the initial response to neuronal stimulation and serve as substrates for second messenger-regulated kinase signalling cascades. The mitogen-activated protein kinase ERK conveys signalling from the synapse to the nucleus but its genetic signature following neuronal activity has not been revealed. The goal of the present study was to identify ERK dependent and independent activity regulated transcriptional programs in the murine hippocampus. We used generalized seizures combined with the pharmacological intervention of MEK activation as an in vivo model to determine the complete transcriptional program initiated by ERK after neuronal activity. Our survey demonstrates that the induction of a large number of activity-regulated genes, including Arc/Arg3.1, Arl5b, Gadd45b, Homer1, Inhba and Zwint, is indeed dependent on ERK phosphorylation. In contrast, expression of a small group of genes, including Npas4, Arl4d, Errfi1, and Rgs2, is only partially dependent or completely independent (Ppp1r15a) of this signalling pathway. Among the identified transcripts are long non-coding (lnc) RNAs and induction of LincPint and splice variants of NEAT1 are ERK dependent. Our survey provides a comprehensive analysis of the transcriptomic response conveyed by ERK signalling in the hippocampus.


Assuntos
MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Regulação da Expressão Gênica , Hipocampo/metabolismo , Sistema de Sinalização das MAP Quinases , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Ativação Transcricional , Processamento Alternativo , Animais , Giro Denteado/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Hibridização In Situ , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Camundongos , Neurônios/metabolismo , Fosforilação , Inibidores de Proteínas Quinases/farmacologia , RNA Longo não Codificante/genética , Convulsões/tratamento farmacológico , Convulsões/etiologia , Convulsões/metabolismo , Convulsões/fisiopatologia , Transcriptoma
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