Loss of microRNA-128 promotes cardiomyocyte proliferation and heart regeneration [miRNA-seq]
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ABSTRACT: To identify the potential microRNAs (miRNAs) involved in the regulation of cardiomyocyte (CM) proliferation during homeostasis and injury, RNA sequencing (RNA-seq) in mouse cardiac ventricles was performed on postnatal day 1, 7, and 28 (P1, P7, and P28). Significant upregulation of MiR-128 was found in P7 hearts as compared to P1. To further specify the effect of miR-128 in the heart, RNA-Seq was performed in control mice (Ctrl) and miR-128 overexpression mice (miR-128OE) on P7. These data provide novel insights into the mechanisms by which adult CMs exit the cell cycle arrest and is fundamental for therapeutic manipulation to stimulate endogenous CM proliferate in cardiac regeneration.
Project description:To identify the potential microRNAs (miRNAs) involved in the regulation of cardiomyocyte (CM) proliferation during homeostasis and injury, RNA sequencing (RNA-seq) in mouse cardiac ventricles was performed on postnatal day 1, 7, and 28 (P1, P7, and P28). Significant upregulation of MiR-128 was found in P7 hearts as compared to P1. To further specify the effect of miR-128 in the heart, RNA-Seq was performed in control mice (Ctrl) and miR-128 overexpression mice (miR-128OE) on P7. These data provide novel insights into the mechanisms by which adult CMs exit the cell cycle arrest and is fundamental for therapeutic manipulation to stimulate endogenous CM proliferate in cardiac regeneration.
Project description:To profile gene expression differences in astrocytes in postnatal cerebellar astrocytes . Comparing cerebellar astrocyte transcriptomes at postnatal days 1, 7, 14, and 28 (P1, P7, P14, and P28),
Project description:MicroRNAs (miRNAs) are small non-coding RNAs that modulate gene expression by negatively regulating translation of target genes. miRNAs involvement in vascular smooth muscle cell (VSMC) biology has been deeply investigated, nevertheless the role of miR-128 in such context has not been yet explored. We aimed to evaluate whether this miR-128 might modulate VSMC phenotype similarly to what was previously observed for other type of contractile cells, such as cardiomyocytes (CM) and skeletal muscle cells. To this end, we compared the expression profiles of miR-128 overexpression in Primary mouse VSMCs and control groups (treated with scrambled miRNA). The expression profiles were defined by Illumina arrays.
Project description:We identify a brain-specific microRNA—miR-128—that represses Nonsense Mediated mRNA Decay (NMD) and thereby controls batteries of transcripts in neural cells. miR-128 represses NMD by targeting the RNA helicase UPF1 and the exon-junction complex core component MLN51. We employed exon arrays for this analysis, as this platform detects expression levels of individual exons and thus allows detection of not only differentially expressed transcripts (DETs), but also alternative isoform transcripts (AITs). The latter is particularly relevant to our study because alternative RNA processing events (e.g., RNA splicing, alternative promoter usage, and alternative polyadenylation-site usage) often place a translation termination codon in a premature context and thereby trigger NMD. Compare different expressed genes and transcript isoforms in mouse NSCs miR-128 positive vs miR-128 negative (Control). Mouse NSCs, that do not normally express miR-128, were nucleofected with either miR-128 (biological triplicates were analysed) or miR-Control (biological triplicates). RNA was extracted 72hrs later, and Exon Array was performed to identify target genes.
Project description:The heart of a newborn mouse has an exceptional capacity to regenerate from myocardial injury but lose it after a week of life, which has been utilized as a valuable model to explore the cues for heart regeneration. More and more researches indicated that glycoprotein played an important role in cardiac regeneration. Elucidating the glycosylation processes associated with heart regeneration will be beneficial for the molecular mechanism studies of heart regeneration as well as discovery of potential therapeutic strategies for human cardiac diseases. In this work, an integrated glycoproteomic and proteomic analysis were performed to investigate the differences in glycoprotein abundances and site-specific glycosylation occupancy between neonatal day 1 (P1) and day 7 (P7) of mouse hearts. The intact glycoepeptides were enriched and identified in both P1 and P7 hearts. To screen for differentially regulated glycoproteins, we compared the expression levels of intact glycopeptides between P1 and P7 hearts using label free quantification. Eventually, the glycosylation occupancy of site-specific N-glycans were obtained by comparing the alterations of intact glycopeptides with their corresponding protein expression levels obtained from global proteomic analysis. These altered glycosylation patterns among proteins between P1 and P7 mouse hearts have a significant potential to aid our understanding of the regenerative capacity loss in neonatal mouse hearts during the first week, thus leading to novel therapeutic approaches to recover the capacity.
Project description:MicroRNA regulates protein expression of cells by repressing translation of specific target messenger transcripts. Loss of the neuron specific microRNA miR-128 in Dopamine D1-receptor expressing neurons in the murine striatum (D1-MSNs) lead to increased neuronal excitability, locomotor hyperactivity and fatal epilepsy. To examine expression changes in the absence of miR-128 in D1-MSNs, we used mice expressing EGFP-tagged ribosomes in D1-MSNs with either D1-MSN-specific homozygous deletion of miR-128-2 locus or no deletion. Transcripts co-immunoprecipitated with tagged ribosomes were analyzed by microarray.
Project description:MicroRNA regulates protein expression of cells by repressing translation of specific target messenger transcripts. Loss of the neuron specific microRNA miR-128 in Dopamine D1-receptor expressing neurons in the murine striatum (D1-MSNs) lead to increased neuronal excitability, locomotor hyperactivity and fatal epilepsy. To examine expression changes in the absence of miR-128 in D1-MSNs, we used mice expressing EGFP-tagged ribosomes in D1-MSNs with either D1-MSN-specific homozygous deletion of miR-128-2 locus or no deletion. Transcripts co-immunoprecipitated with tagged ribosomes were analyzed by microarray. 9 mutant animals ( D1-MSN-tagged ribosome; D1-MSN specific miR-128-2 homozygous deletion) and 7 age matched littermate control animals (D1-MSN-tagged ribosome only).
Project description:This experiment is designed to evaluate gene expression alteration and significant pathway(s) following miR-128 transduction in A549 lung cancer cells. We find several significant pathways, including the Wnt/β-catenin signaling and TGF-β signaling activated by miR-128 overexpression.
Project description:The experiment was deigned to identify the genes which get altered after the over expression of hsa-miR-128 in HEK293T cells. The HEK293T cells plated in 6-well plate were transfected with 4 micrograms of cloned miR-128 (p128) in three biological replicates.
Project description:In order to identify cell-autonomous factors that might be regulating the neurogenic niche, type 1 and 2 progenitors were FACS sorted from P7 and P28 animals and their cDNA was hybridized to a stem cell-specific microarray. Keywords: differential gene expression analysis Two samples per microarray (P7 and P28). Each microarray experiment was done in triplicate. P7 is the control sample.