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Characterization of miRNA transcriptome of skeletal muscle from broiler and layer chicken lines by deep sequencing

ABSTRACT: In the current study, we expanded our previous work to identify miRNAs implicated in the myogenesis regulation through the comparison of miRNAs transcriptome in skeletal muscle tissues between broilers and layers. To address that, we firstly performed Solexa deep sequencing to profile miRNAs expressed in chicken skeletal muscle tissues. Sequence tags analyses not only enable us to report a group of highly abundant known miRNAs expressed in skeletal muscles but most importantly to identify novel putative chicken miRNAs from skeletal muscle tissue. Further miRNA transcriptome comparison and real-time RT-PCR validation experiments revealed seveal differentially expressed miRNAs between broilers and layers.

ORGANISM(S): Gallus gallus

PROVIDER: GSE20942 | GEO | 2011/03/18

SECONDARY ACCESSION(S): PRJNA129643

REPOSITORIES: GEO

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Project description:The genetic closeness and divergent muscle growth rates of broilers and layers make them great models for myogenesis study. In order to discover the molecular mechanisms determining the divergent muscle growth rates and muscle fiber sizes in different chicken lines, we systematically identified differentially expressed genes between broilers and layers during muscle development (postnatal 1 day, 2 weeks, 4 weeks, 6 weeks and 8 weeks) by microarray hybridization experiment. Taken together, 543 differentially expressed probe sets were found between broilers and layers across different developmental stages, including genes related to muscle growth and hypertrophy, fatty acid transportation and metabolism, protein degradation, and several important signaling pathways. The expression profiles of a few differentially expressed genes were highly correlated with the growth rates of broilers and layers. We also identified SNPs within upstream transcription factor binding sites of a few differentially expressed genes, indicating that these SNPs might be the causing factor of the expression differences of these genes between broilers and layers. These studies provided new clues for deciphering mechanisms underlining muscle development and organ size control in different chicken lines, will shed light on the study of human muscle related disease as well. Experiment Overall Design: Pectoralis major muscles were sampled from broilers and layers at indicated developmental stage (postnatal 1 day, 2 weeks, 4 weeks, 6 weeks and 8 weeks). Trizol extraction of total RNA was performed according to the manufacturer's instructions. Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 2 ug total RNA. Affymetrix Gene Chip array hybridization was carried out according to Affymetrix Expression Analysis Technical Manual by GeneTech Biotechnology Limited Company (Shanghai, China). Following labeled cRNA fragmentation, 15 ug of cRNA were hybridized for 16 hr at 45â on Affymetrix Chicken Genome Array. After standard washing and staining, GeneChips were scanned using the Affymetrix GeneChip Scanner 7G.

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Characterization of miRNA transcriptome of skeletal muscle from broiler and layer chicken lines by deep sequencing

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