title: PPARγ is regulated by miR-27b-3p negatively and plays an important role in porcine oocyte maturation.
abstract: To elucidate the key miRNAs and the signalling pathways that are involved in porcine oocyte maturation, we performed a deep sequencing analysis of the miRNAs of pig germinal vesicle (GV) oocytes and metaphase II (MII) oocytes. Seven differentially expressed (DE) miRNAs were identified and the expression levels of miR-21 and miR-27b-3p were further confirmed by QPCR analysis. The target genes of 7 DE miRNAs were predicted and subjected to pathway analysis. Interestingly, fatty acid metabolism and fatty acid biosynthesis were the top two significantly enriched molecular functions during oocyte maturation. Heat map, which was built with 7 DE miRNAs and the enriched the molecular functions, revealed that miR-21, miR-27b-3p, miR-10a-5p and miR-10b-5p were involved in fatty acid metabolism. In particular, the regulatory role of miR-27b-3p on peroxisome proliferator-activated receptor-γ (PPARγ) was confirmed by their inversed expression patterns in GV and MII oocytes and luciferase report assays. In addition, we observed that PPARγ agonist (rosiglitazone) treatment significantly enhanced porcine oocyte maturation rate and early embryo developmental competent. Taken together, our results demonstrated that miR-27b and its target, PPARγ, play the vital roles in pig oocyte maturation through regulating the fatty acid metabolism. These data increased our understanding of the regulatory gene networks in porcine oocyte maturation and development.
authors.affiliation: State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, PR China.
authors.forename: Chunlei
authors.initials: C
authors.lastname: Song
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100101, PR China; Cunji Medical School, University of Chinese Academy of Sciences, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yao
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100101, PR China; Cunji Medical School, University of Chinese Academy of Sciences, PR China.
authors.forename: Chunwei
authors.initials: C
authors.lastname: Cao
authors.affiliation: State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, PR China.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100101, PR China; Cunji Medical School, University of Chinese Academy of Sciences, PR China.
authors.forename: Jiaojiao
authors.initials: J
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100101, PR China; College of Life Science, Hebei University, 071000, PR China.
authors.forename: Ziqiang
authors.initials: Z
authors.lastname: Han
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100101, PR China; Cunji Medical School, University of Chinese Academy of Sciences, PR China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100101, PR China; Cunji Medical School, University of Chinese Academy of Sciences, PR China.
authors.forename: Guosong
authors.initials: G
authors.lastname: Qin
authors.affiliation: State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, PR China.
authors.forename: Cong
authors.initials: C
authors.lastname: Tao
authors.affiliation: Rongchang Breeding Company, Shandong, Binzhou, 251900, PR China.
authors.forename: Chengbo
authors.initials: C
authors.lastname: Li
authors.affiliation: Tsinghua University High School, Beijing, 100094, PR China.
authors.forename: Haoran
authors.initials: H
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100101, PR China; Cunji Medical School, University of Chinese Academy of Sciences, PR China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, PR China.
authors.forename: Kui
authors.initials: K
authors.lastname: Li
authors.affiliation: State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, PR China. Electronic address: wangyanfang@caas.cn.
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Wang
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D016000:Cluster Analysis; D005227:Fatty Acids / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D059471:In Vitro Oocyte Maturation Techniques; D035683:MicroRNAs / Q000145:classification / Q000235:genetics*; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D047495:PPAR gamma / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077154:Rosiglitazone; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction / Q000235:genetics; D013552:Swine; D045162:Thiazolidinediones / Q000494:pharmacology
chemical_list: D020413:3' Untranslated Regions; D005227:Fatty Acids; D035683:MicroRNAs; D047495:PPAR gamma; D045162:Thiazolidinediones; D000077154:Rosiglitazone
keywords: Fatty acid metabolism; Oocyte maturation; PPARγ; Pig; miRNA