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Equine tissue miRNA profiling

ABSTRACT: Purpose: Identification of miRNA expression profiles of selected horse tissues. Methods: miRNA-seq analysis was performed on lung, heart and liver samples collected from 4 horses. The miRNA libraries were constructed from total RNA using NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs) according to the manufacturer protocol. The quantification of the obtained libraries was performed on a Qubit 2.0 spectrophotometer (Invitrogen, Life Technologies), while a quality control on a TapeStation 2200 instrument (D1000 ScreenTape; Agilent). 75 single-end cycle sequencing was performed on the NextSeq 500/550 platform (Illumina) with the use of NextSeq High Output Reagents v2 (Illumina). Results: The comparison of miRNA profiles between the investigated tissues revealed 185 differentially expressed microRNAs (p adj≤0.05) in the heart samples versus the liver samples, 172 DE miRNAs (p adj ≤0.05) in the lung samples in comparison to the liver samples, and 155 in the lung samples versus the heart samples. Conclusions: Obtained results show varied miRNA expression profiles characteristics for each investigated hore tissue.

ORGANISM(S): Equus caballus

PROVIDER: GSE202502 | GEO | 2023/06/01

REPOSITORIES: GEO

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Project description:Purpose: The aim of present research was to identify miRNAs potentially related with muscle growth in pig Methods: miRNA-seq analysis was performed on longissimus lumborum samples collected from 13 pigs representing two sire-line breeds – Pietrain and Hampshire. Based on dissection data, pigs were selected from a larger population in terms of weight of loin muscle to obtain the most extreme groups in each breed The miRNA libraries were constructed form total RNA with the use Neb Next Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs) according to the protocol. The quantification of obtained libraries was performed on a Qubit 2.0 spectrophotometer (Invitrogen, Life Technologies) and a TapeStation 2200 instrument (D1000 ScreenTape; Agilent). The miRNA-seq was performed in 36 single-end cycles on HiScanSQ platform (Illumina) with the use of TruSeq SR Cluster Kit v3- CBOT-HS and TruSeq SBS Kit v 3 - HS (Illumina). Differentially expressed microRNAs were detected using the DESeq2 software while validation was performed by qPCR. Results: miRNA-seq approach allowed to identify 196 known and 149 potentially novel miRNAs in both breeds. The comparison of miRNA profiles between pig groups with different loin weight showed differential expression of 57 microRNAs for Hampshire pigs and 34 miRNAs for Pietrain pigs. Furthermore, 18 common microRNAs with differential expression were identified for both breeds tested. The Gene Ontology analysis (DIANA mirPath v.3 tool) confirmed that detected genes were involved in regulation of cell cycle, fatty acid biosynthesis, regulation of actin cytoskeleton, lysine degradation and ubiquitin mediated proteolysis. According to the KEGG Database, these DE miRNAs enrich the following pathways, i.a.: the Hippo signaling pathway, TGF-beta signaling pathway and PI3K-Akt signaling pathway. Conclusions: Obtained results allow to propose the panel of miRNAs, which can be related with porcine muscle growth and development. Such analysis can be the basis of future research in terms of identification of candidate miRNAs related with important production traits in pigs.

Project description:Purpose: The aim of the study was to compare the miRNA expression in non-infected (H) mammary gland parenchyma samples with that of glands infected with coagulase-positive staphy lococci (CoPS) or coagulase-negative staphylococci (CoNS) using next-generation sequencing. Methods: miRNA-seq analysis was performed on mammary gland parenchyma samples collected from non-infected cows and those infected with coagulase-positive or -negative staphylococci. The miRNA libraries were constructed from total RNA using NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs) according to the manufacturer protocol. The quantification of the obtained libraries was performed on a Qubit 2.0 spectrophotometer (Invitrogen, Life Technologies), while a quality control on a TapeStation 2200 instrument (D1000 ScreenTape; Agilent). Single-end cycle sequencing was performed on the HiScanSQ platform (Illumina) with the use of TruSeq SR Cluster Kit v3- CBOT-HS and TruSeq SBS Kit v 3 - HS (Illumina). MicroRNA differentially expressed between investigated groups were identified with the DESeq2 software. Results: Comparing the CoPS and H groups, 256 known and 260 potentially new miRNAs were identifed, including 32 that were diferentially expressed (p≤0.05), of which 27 were upregulated and 5 downregulated. Comparing the CoNS and H groups, 242 known and 171 new unique miRNAs were identifed: 10 were upregulated (p≤0.05), and 2 downregulated (p≤0.05). Comparing CoPS with H and CoNS with H, 5 Kyoto Encyclopedia of Genes and Genomes pathways were identifed; in both comparisons, diferentially-expressed miRNAs were associated with the bacterial invasion of epithelial cells and focal adhesion pathways. Four gene ontology terms were identifed in each comparison, with 2 being common to both immune system processes and signal transduction. Conclusions: Obtained results enabled us to characterize the miRNA profile of the mammary gland parenchyma tissue, not only the healthy one but also the tissue infected with coagulase-positive and negative staphylococci as well as allowed identification of miRNAs differing the examined groups and characteristic for the staphylococci infection. They also indicated that miRNAs, especially miR-99 and miR-182, play an essential role in the epigenetic regulation of a range of cellular processes, including immunological systems bacterial growth in dendritic cells and disease pathogenesis (miR-99), DNA repair and tumor progression (miR-182).

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Equine tissue miRNA profiling

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