Sequencing libraries were generated using the TruSeq Small RNA Library Prep Kit (Illumina, San Diego, CA, USA), following the manufacturer’s recommendations. 5c). 2011;7(7):1045–55. ML, DQX, HSD, and YBY conceived this study; HSD and CFZ performed the experiments. de Kretser DM, Loveland KL, Meinhardt A, Simorangkir D, Wreford N. Spermatogenesis. Meishan pigs were domesticated nearly 5000 years ago in the Taihu Lake region of China and are one of the rarest pigs in not just the United States, but in the world. In this study, we obtained comprehensive sRNA-seq data and modelled the miRNA-mRNA regulatory networks related to testis development and spermatogenesis through the integrated analysis of differentially expressed miRNAs and target mRNAs between Duroc and Meishan boars. MicroRNAs activate gene transcription epigenetically as an enhancer trigger. McCoard SA, Wise TH, Lunstra DD, Ford JJ. As a class of endogenous small (~ 22 nucleotides) noncoding RNAs, miRNAs mediate post-transcriptional gene expression in animals and fine-tune the expression of approximately 30% of all mammalian protein-coding genes [4,5,6]. Kasimanickam V, Kastelic J. MicroRNA in sperm from Duroc, Landrace and Yorkshire boars. Primers of genes and miRNAs used for quantitative real-time PCR. GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes. These complex networks identified in our study may participate in regulating boar fertility. Bao J, Li D, Wang L, Wu J, Hu Y, Wang Z, et al. Four significantly differentially expressed miRNAs and their predicted target genes were validated by quantitative real-time polymerase chain reaction, and phospholipase C beta 1 (PLCβ1) gene was verified to be a target of ssc-mir-423-5p. The expression levels of ssc-mir-423-5p were analyzed by qRT-PCR. PubMed Central The Meishan is a breed of domestic pig native to Southern China that is considered a sub-group of the Taihu pigs. Promoter RNA links transcriptional regulation of inflammatory pathway genes. 2017;14(10):1326–34. NC, negative control. Identification and differential expression of microRNAs in testis and ovary of Amur sturgeon (Acipenser schrenckii). These miRNAs may serve potential roles in regulating testis development, and their specific expressions may induce the differences in sexual maturity and spermatogenesis. They are a "red meat" pig, prized for their lard and flavorful meat. Cell Cycle. 5d-g, Additional file 5: Figure S2, Additional file 6: Figure S3). PubMed 2009;87(8):2536–43. The qRT-PCR of the miRNAs and mRNAs was performed by using a standard UltraSYBR Mixture (CWBIO, Beijing, China) in the Roche LightCyler 480 system (Roche, Mannheinm, Germany) according to the manufacturer’s instructions. RNA quality and quantity were measured using a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). The number of genes detected in the six samples. Then the cut membranes were blocked with 5% non-fat dried milk and incubated overnight with primary antibodies specific for PLCβ1 (1:1000; ABclonal, USA, A1971). A total of 1,078,105 and 878,097 unique sRNA from Duroc and Meishan boar testes were mapped to the porcine reference genome (Sscrofa10.2), respectively (Table 2). Article Shu mai (USDA), Moo Shu (Illinois-Iowa), Kung Pao(Iowa-Illinois) and General Tso (USDA). Qiu W, Zhu Y, Wu Y, Yuan C, Chen K, Li M. Identification and expression analysis of microRNAs in medaka gonads. Acta Biochim Biophys Sin. miR-34c inhibits proliferation and enhances apoptosis in immature porcine Sertoli cells by targeting the SMAD7 gene. A representative miRNA-mRNA regulatory network of biological pathways was shown in Fig. Ding H, Luo Y, Liu M, Huang J, Xu D. Histological and transcriptome analyses of testes from Duroc and Meishan boars. 2007;23:175–205. Google Scholar. Springer Nature. Meishan pigs have recently been added to the Conservation Priority List of The Livestock Conservancy, meaning the breed is just now able to bring on new stewards to help. The porcine PLCβ1 3’UTR was amplified with primers PLCβ1–3’UTR-F/PLCβ1–3’UTR-R in Additional file 8: Table S5, and their products were double-digested with Sac I (Takara, Dalian, China) and Xho I (Takara, Dalian, China), and then was cloned into the pmirGLO vector (Promega, Madison, WI, USA) as a wild-type plasmid. Clip domain serine protease and its homolog respond to Vibrio challenge in Chinese white shrimp, Fenneropenaeus chinensis. Repertoire of porcine microRNAs in adult ovary and testis by deep sequencing. Ran M, Luo H, Gao H, Tang X, Chen Y, Zeng X, et al. Sertoli cell differentiation in pubertal boars. Primers for amplification of porcine PLCβ1 3’UTR. The diameter and number of seminiferous tubules determine the onset of puberty in males [17]. The number of genes involved in KEGG pathway and GO term. Mol Reprod Dev. Meishan Pig. Then the equal amount of RNA from three individuals of the same age and breed were mixed to form 6 RNA pools, named D20, D75, D270, M20, M75, and M270. And the wild-type plasmid was used as template to construct mutated plasmid with primers PLCβ1–3’UTR-F/PLCβ1–3’UTR-MR in Additional file 8: Table S5, and the primer PLCβ1–3’UTR-R contained the binding site of ssc-mir-423-5p. Sows grow to be about 57.8 cm high, 100 cm around the chest, and 61.6 kg in live-weight. 2a). Effect of dietary Astragalus polysaccharide supplements on testicular miRNA expression profiles and enzymatic changes of breeder cocks. Small RNA profiling and characterization of piRNA clusters in the adult testes of the common marmoset, a model primate. Significant differences were evaluated through an independent-sample t-test. The number of miRNA/mRNA-negative pairs between Duroc and Meishan boars at 75-day time point was obviously higher than that at 20- and 270-day time points, and more negative pairs were detected from 20 to 75 days in Meishan boars than those in Duroc boars (Fig. PubMed Central Xiao M, Li J, Li W, Wang Y, Wu FZ, Xi YP, et al. Hirano T, Iwasaki YW, Lin ZY, Imamura M, Seki NM, Sasaki E, et al. 2009;26(5):787–98. The statistics for miRNA/mRNA-regulation pairs in Duroc and Meishan boars. Int J Mol Sci. Curry E, Ellis SE, Pratt SL. Integrated analysis between mRNA and miRNA libraries was performed using miRTrail bioinformatics tool. The Meishan is a breed of domestic pig named for the Chinese prefecture of Meishan.
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