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| Status |
Public on Feb 08, 2011 |
| Title |
Global profiling of gene expression in mouse astrocyte in response to the potential longevity determinant miR-29 |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
MicroRNAs (miRNAs) are an evolutionarily conserved large class of small non-coding RNAs that mediate post-transcriptional silencing of genes and influence a broad spectrum of biological processes ranging from embryonic development to organismal death. Our previous study identified the miR-29 family, three paralogous species of miR-29a/b/c, as the most predominantly expressed small RNA in aged mouse brain compared to the neonate one. Mouse brain miR-29 is highly astrocytic. Its expression is quiescent during early brain development, and then steadily increases to a plateau state around reproductive maturity. To explore the functional relevance of miR-29 expression in early life to the neural physiology of the mouse brain from the mechanistic perspective of mammalian species-specific lifespan, we here undertook a gain-of-function approach through exogenous expression of miR-29 in astrocyte from mouse fetus and surveyed the resulting alteration in both the transcriptional and translational levels. DNA microarray analyses retrieved a total of 5,589 genes showing temporal significant expression changes in the miR-29-transfected fetal astrocytes, and classified them into two gene groups: positively or negatively regulated by miR-29. Mass spectrometry (MS)-based quantification of translational products of miR-29-responsive genes identified 18 species of miR-29 target candidates. We performed functional enrichment analyses utilizing bioinformatics resources to characterize the gene sets thus identified, and found their expression trend that favor the processes for facilitating cell differentiation while supporting normal cell proliferation /survival, which is somewhat different from the functional signatures of miR-29 as observed at adult stages, implying a pleiotropic property of miR-29 depending on the developmental context. Our present results strongly suggest that miR-29 in the developing mouse brain serves as the central coordinator to shift the global gene expression toward adult phenotypes, through which ensure the programmed transition in the life course to the post-developmental/reproductive stage which has inherently been set to delimit the mouse life potential.
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| Overall design |
DNA microarray experiment: Sample labeling, hybridization and washing were performed following the standard protocol detailed in the Agilent One-Color Microarray-Based Gene Expression Analysis ver.5.7. Briefly, a One-color Spike-Mix was diluted 1000-fold and a 5-μl aliquot of the diluted mix were added to every 0.5 μg of total RNA samples (three each of extraction replicates of miR-29-treated sample and control sample named elsewhere) prior to labeling reaction. The labeling reaction was carried out separately for the RNA samples using a Quick Amp Labeling Kit one-color, in the presence of cyanine 3-CTP. The dye-labeled target (1.6 μg as cRNA) was fragmented and hybridized on an Agilent Whole Mouse Genome 4X44K microarray at 65°C for 17 hr with a Gene Expression Hybridization Kit. The hybridized slide was washed in Gene Expression Wash Buffer 1 at room temperature for 1 min, which was followed by a wash for 1 min in Gene Expression Wash Buffer 2. The processed microarrays were scanned using an Agilent DNA Microarray Scanner. Data extraction from raw image files was done with Agilent Feature Extraction software ver.10.7.
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| Web link |
http://www.nara.kindai.ac.jp/bio/tanabe/miR-29-transfection
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| Contributor(s) |
Tanabe H |
| Citation |
Tanabe, H., Kohno, N. and Yamaguchi, M. 2012. Global profiling of gene expression in mouse astrocyte in response to the potential longevity determinant miR-29. Mem. Fac. Agr. Kinki Univ. 45: 1-16; kurepo.clib.kindai.ac.jp/modules/xoonips/download.php?file_id=7676
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| Submission date |
Feb 02, 2011 |
| Last update date |
May 10, 2018 |
| Contact name |
Hiroyuki Tanabe |
| Organization name |
Kinki University
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| Department |
Faculty of Agriculture Department of Advanced Bioscience
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| Street address |
3327-204 Nakamachi
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| City |
Nara City |
| State/province |
Nara |
| ZIP/Postal code |
631-8505 |
| Country |
Japan |
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| Platforms (1) |
| GPL4134 |
Agilent-014868 Whole Mouse Genome Microarray 4x44K G4122F (Feature Number version) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA137673 |
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