{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T01:07:36Z","timestamp":1789607256695,"version":"build-2803163510"},"reference-count":51,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2024,5,29]],"date-time":"2024-05-29T00:00:00Z","timestamp":1716940800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007129","name":"Natural Science Foundation of Shandong Province","doi-asserted-by":"publisher","award":["ZR2022MC100"],"award-info":[{"award-number":["ZR2022MC100"]}],"id":[{"id":"10.13039\/501100007129","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007129","name":"Natural Science Foundation of Shandong Province","doi-asserted-by":"publisher","award":["31702331"],"award-info":[{"award-number":["31702331"]}],"id":[{"id":"10.13039\/501100007129","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["ZR2022MC100"],"award-info":[{"award-number":["ZR2022MC100"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31702331"],"award-info":[{"award-number":["31702331"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Infrastructure of Fishery Germplasm Resources, China","award":["ZR2022MC100"],"award-info":[{"award-number":["ZR2022MC100"]}]},{"name":"National Infrastructure of Fishery Germplasm Resources, China","award":["31702331"],"award-info":[{"award-number":["31702331"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>DNA methylation is an important way to regulate gene expression in eukaryotes. In order to reveal the role of DNA methylation in the regulation of germ cell-specific piwi gene expression during spermatogenesis of Japanese flounder (Paralichthys olivaceus), the expression profiles of piwil1 (piwi-like 1) and piwil2 (piwi-like 2) genes in the gonads of female, male, and sex-reversed pseudo-male P. olivaceus were analyzed, and the dynamic of DNA methylation was investigated. As a result, piwil1 and piwil2 genes were highly expressed in the testis of both male and pseudo-male P. olivaceus, with significant variation among male individuals. The DNA methylation levels in the promoter regions of both piwil1 and piwil2 were negatively correlated with their expression levels, which may contribute to the transcriptional regulation of piwi genes during spermatogenesis. There was also sperm quality variation among male P. olivaceus, and the sperm curvilinear velocity was positively correlated with the expression of both piwil1 and piwil2 genes. These results indicated that the DNA methylation in piwil1 and piwil2 promoter regions may affect the initiation of piwi gene transcription, thereby regulating gene expression and further affecting the spermatogenesis process and gamete quality in P. olivaceus.<\/jats:p>","DOI":"10.3390\/ijms25115935","type":"journal-article","created":{"date-parts":[[2024,5,30]],"date-time":"2024-05-30T10:42:34Z","timestamp":1717065754000},"page":"5935","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["DNA Methylation Mediates Sperm Quality via piwil1 and piwil2 Regulation in Japanese Flounder (Paralichthys olivaceus)"],"prefix":"10.3390","volume":"25","author":[{"given":"Wenyu","family":"Zong","sequence":"first","affiliation":[{"name":"MOE Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yapeng","family":"Wang","sequence":"additional","affiliation":[{"name":"MOE Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingqun","family":"Zhang","sequence":"additional","affiliation":[{"name":"MOE Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Lu","sequence":"additional","affiliation":[{"name":"MOE Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weigang","family":"Li","sequence":"additional","affiliation":[{"name":"MOE Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fengchi","family":"Wang","sequence":"additional","affiliation":[{"name":"MOE Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8835-3171","authenticated-orcid":false,"given":"Jie","family":"Cheng","sequence":"additional","affiliation":[{"name":"MOE Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China"},{"name":"Key Laboratory of Tropical Aquatic Germplasm of Hainan Province, Sanya Oceanographic Institution, Ocean University of China, Sanya 572024, China"},{"name":"Laboratory for Marine Fisheries Science and Food Production Processes, National Laboratory for Marine Science and Technology, Qingdao 266237, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1146\/annurev.cellbio.24.110707.175327","article-title":"The biogenesis and function of PIWI proteins and piRNAs: Progress and prospect","volume":"25","author":"Thomson","year":"2009","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1126\/science.1130164","article-title":"Characterization of the piRNA complex from rat testes","volume":"313","author":"Lau","year":"2006","journal-title":"Science"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1126\/science.1129333","article-title":"A distinct small RNA pathway silences selfish genetic elements in the germline","volume":"313","author":"Vagin","year":"2006","journal-title":"Science"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1016\/j.cell.2007.01.043","article-title":"Discrete small RNA-generating loci as master regulators of transposon activity in Drosophila","volume":"128","author":"Brennecke","year":"2007","journal-title":"Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1038\/nrg2072","article-title":"Transposable elements and the epigenetic regulation of the genome","volume":"8","author":"Slotkin","year":"2007","journal-title":"Nat. Rev. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2214","DOI":"10.1101\/gad.1454806","article-title":"Specific association of Piwi with rasiRNAs derived from retrotransposon and heterochromatic regions in the Drosophila genome","volume":"20","author":"Saito","year":"2006","journal-title":"Genes Dev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1002\/jcb.23363","article-title":"Piwis and piwi-interacting RNAs in the epigenetics of cancer","volume":"113","author":"Siddiqi","year":"2012","journal-title":"J. Cell. Biochem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1016\/j.cell.2009.01.045","article-title":"Small RNAs as guardians of the genome","volume":"136","author":"Malone","year":"2009","journal-title":"Cell"},{"key":"ref_9","first-page":"683","article-title":"The regulatory functions of piRNA\/PIWI in spermatogenesis","volume":"39","author":"Yuan","year":"2017","journal-title":"Yi Chuan Hered."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1016\/j.stem.2018.08.007","article-title":"Single-cell RNA sequencing analysis reveals sequential cell fate transition during human spermatogenesis","volume":"23","author":"Wang","year":"2018","journal-title":"Cell Stem Cell"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.cell.2007.03.026","article-title":"A role for Piwi and piRNAs in germ cell maintenance and transposon silencing in Zebrafish","volume":"129","author":"Houwing","year":"2007","journal-title":"Cell"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2702","DOI":"10.1038\/emboj.2008.204","article-title":"Zili is required for germ cell differentiation and meiosis in zebrafish","volume":"27","author":"Houwing","year":"2008","journal-title":"EMBO J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1016\/j.bbrc.2011.12.062","article-title":"Specific expression of Olpiwi1 and Olpiwi2 in medaka (Oryzias latipes) germ cells","volume":"418","author":"Zhao","year":"2012","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.cbpb.2018.03.005","article-title":"Differential expression of two Piwil orthologs during embryonic and gonadal development in pufferfish, Takifugu fasciatus","volume":"219","author":"Wen","year":"2018","journal-title":"Comp. Biochem. Physiol. B Biochem. Mol. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1007\/s00427-016-0554-3","article-title":"Genomic identification, rapid evolution, and expression of Argonaute genes in the tilapia, Oreochromis niloticus","volume":"226","author":"Tao","year":"2016","journal-title":"Dev. Genes Evol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.gene.2018.03.016","article-title":"Piwil1 gene is regulated by hypothalamic-pituitary-gonadal axis in turbot (Scophthalmus maximus): A different effect in ovaries and testes","volume":"658","author":"Wang","year":"2018","journal-title":"Gene"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.margen.2014.04.004","article-title":"Cloning, expression and methylation analysis of piwil2 in half-smooth tongue sole (Cynoglossus semilaevis)","volume":"18","author":"Zhang","year":"2014","journal-title":"Mar. Genom."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1617","DOI":"10.1111\/j.1742-4658.2008.06330.x","article-title":"Epigenetics: Differential DNA methylation in mammalian somatic tissues","volume":"275","author":"Nagase","year":"2008","journal-title":"FEBS J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1038\/ng1089","article-title":"Epigenetic regulation of gene expression: How the genome integrates intrinsic and environmental signals","volume":"33","author":"Jaenisch","year":"2003","journal-title":"Nat. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"458","DOI":"10.1210\/en.2012-1610","article-title":"Changes in histone modification and DNA methylation of the StAR and Cyp19a1 promoter regions in granulosa cells undergoing luteinization during ovulation in rats","volume":"154","author":"Lee","year":"2013","journal-title":"Endocrinology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.ygcen.2010.07.012","article-title":"Tissue-specific promoter methylation coincides with Cyp19 gene expression in buffalo (Bubalus bubalis) placenta of different stages of gestation","volume":"169","author":"Ghai","year":"2010","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2881","DOI":"10.1210\/en.2012-2220","article-title":"Epigenetic modifications during sex change repress gonadotropin stimulation of cyp19a1a in a teleost ricefield eel (Monopterus albus)","volume":"154","author":"Zhang","year":"2013","journal-title":"Endocrinology"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Navarro-Mart\u00edn, L., Vinas, J., Ribas, L., Diaz, N., Gutierrez, A., Di Croce, L., and Piferrer, F. (2011). DNA methylation of the gonadal aromatase (cyp19a) promoter is involved in temperature-dependent sex ratio shifts in the European sea bass. PLoS Genet., 7.","DOI":"10.1371\/journal.pgen.1002447"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Su, J., Yi, S., Gao, Z., Abbas, K., and Zhou, X. (2023). DNA methylation mediates gonadal development via regulating the expression levels of cyp19a1a in loach Misgurnus anguillicaudatus. Int. J. Biol. Macromol., 235.","DOI":"10.1016\/j.ijbiomac.2023.123794"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"437067","DOI":"10.3389\/fgene.2019.00522","article-title":"Transcriptome profiling insights the feature of sex reversal induced by high temperature in tongue sole Cynoglossus semilaevis","volume":"10","author":"Liu","year":"2019","journal-title":"Front. Genet."},{"key":"ref_26","first-page":"473","article-title":"Cloning of Sox9 gene and the relationship between CpG island methylation and gene expression in Culter alburnus","volume":"43","author":"Jia","year":"2019","journal-title":"Acta Hydrobiol. Sin."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"113","DOI":"10.4161\/epi.27237","article-title":"Epigenetic loss of the PIWI\/piRNA machinery in human testicular tumorigenesis","volume":"9","author":"Ferreira","year":"2014","journal-title":"Epigenetics"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Heyn, H., Ferreira, H.J., Bassas, L., Bonache, S., Sayols, S., Sandoval, J., Esteller, M., and Larriba, S. (2012). Epigenetic disruption of the PIWI pathway in human spermatogenic disorders. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0047892"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"8317","DOI":"10.1007\/s11033-020-05843-3","article-title":"Characterization of sexual size dimorphism and sex-biased genes expression profile in the olive flounder","volume":"47","author":"Ryu","year":"2020","journal-title":"Mol. Biol. Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1016\/j.semcdb.2008.12.002","article-title":"Sex determination in flatfishes: Mechanisms and environmental influences","volume":"20","author":"Luckenbach","year":"2009","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Song, H., Xing, C., Lu, W., Liu, Z., Wang, X., Cheng, J., and Zhang, Q. (2019). Rapid evolution of piRNA pathway and its transposon targets in Japanese flounder (Paralichthys olivaceus). Comp. Biochem. Physiol. D Genom. Proteom., 31.","DOI":"10.1016\/j.cbd.2019.100609"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Wang, B., Wang, H., Song, H., Jin, C., Peng, M., Gao, C., Yang, F., Du, X., Qi, J., and Zhang, Q. (2019). Evolutionary significance and regulated expression of Tdrd family genes in gynogenetic Japanese flounder (Paralichthys olivaceus). Comp. Biochem. Physiol. D Genom. Proteom., 31.","DOI":"10.1016\/j.cbd.2019.05.003"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1007\/s42995-021-00118-7","article-title":"Evolutionary dynamics and conserved function of the Tudor domain-containing (TDRD) proteins in teleost fish","volume":"4","author":"Liu","year":"2022","journal-title":"Mar. Life Sci. Technol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.gene.2019.03.045","article-title":"Roles of piwil1 gene in gonad development and gametogenesis in Japanese flounder, Paralichthys olivaceus","volume":"701","author":"Ni","year":"2019","journal-title":"Gene"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1007548","DOI":"10.3389\/fgene.2022.1007548","article-title":"Y-specific amh allele, amhy, is the master sex-determining gene in Japanese flounder Paralichthys olivaceus","volume":"13","author":"Hattori","year":"2022","journal-title":"Front. Genet."},{"key":"ref_36","unstructured":"Zhang, B. (2020). Identification and Application of Sex Markers and Sex-Related Gene in Pseudomale Cynoglossus semilaevis. [Ph.D. Thesis, Shanghai Ocean University]."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Zhang, B., Zhao, N., Jia, L., Che, J., He, X., Liu, K., and Bao, B. (2020). Identification and application of piwi-interacting RNAs from seminal plasma exosomes in Cynoglossus semilaevis. BMC Genom., 21.","DOI":"10.1186\/s12864-020-6660-7"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1007\/s10126-022-10109-z","article-title":"Comparative piRNAs profiles give a clue to transgenerational inheritance of sex-biased piRNAs in Cynoglossus semilaevis","volume":"24","author":"Zhao","year":"2022","journal-title":"Mar. Biotechnol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3527","DOI":"10.1210\/en.2010-1469","article-title":"Proteolytically activated, recombinant anti-M\u00fcllerian hormone inhibits androgen secretion, proliferation, and differentiation of spermatogonia in adult zebrafish testis organ cultures","volume":"152","author":"Skaar","year":"2011","journal-title":"Endocrinology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1007\/s10142-018-0595-y","article-title":"Transcriptomics analysis revealing candidate networks and genes for the body size sexual dimorphism of Chinese tongue sole (Cynoglossus semilaevis)","volume":"18","author":"Wang","year":"2018","journal-title":"Funct. Integr. Genom."},{"key":"ref_41","unstructured":"Li, W. (2012). Studies on the Artificial Induction of Polyploidy and Its Effect on Growth and Development of Triploid in Half-Smooth Tongue Sole (Cynoglossus semilaevis). [Master\u2019s Thesis, Shanghai Ocean University]."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1093\/humupd\/dmp048","article-title":"World Health Organization reference values for human semen characteristics","volume":"16","author":"Cooper","year":"2010","journal-title":"Hum. Reprod. Update"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1007\/s10126-011-9395-2","article-title":"Induction of mitogynogenetic diploids and identification of WW super-female using sex-specific SSR markers in half-smooth tongue sole (Cynoglossus semilaevis)","volume":"14","author":"Chen","year":"2012","journal-title":"Mar. Biotechnol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1038\/s41437-018-0126-6","article-title":"New locus reveals the genetic architecture of sex reversal in the Chinese tongue sole (Cynoglossus semilaevis)","volume":"121","author":"Cui","year":"2018","journal-title":"Heredity"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1457","DOI":"10.1007\/s10695-018-0505-1","article-title":"Subjective and objective assessment of fish sperm motility: When the technique and technicians matter","volume":"44","author":"Gallego","year":"2018","journal-title":"Fish Physiol. Biochem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1982","DOI":"10.1210\/en.2018-00015","article-title":"An miR-200 cluster on chromosome 23 regulates sperm motility in zebrafish","volume":"159","author":"Xiong","year":"2018","journal-title":"Endocrinology"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1104","DOI":"10.1071\/RD18453","article-title":"Male reproductive dysfunction in Solea senegalensis: New insights into an unsolved question","volume":"31","author":"Riesco","year":"2019","journal-title":"Reprod. Fertil. Dev."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1007\/s10815-016-0682-8","article-title":"The role of epigenetics in idiopathic male infertility","volume":"33","author":"Gunes","year":"2016","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Woods, L.C., Li, Y., Ding, Y., Liu, J., Reading, B.J., Fuller, S.A., and Song, J. (2018). DNA methylation profiles correlated to striped bass sperm fertility. BMC Genom., 19.","DOI":"10.1186\/s12864-018-4548-6"},{"key":"ref_50","unstructured":"Sun, Z. (2008). Histological Studies on Gonadal Ontogenesis, Differentiation and Development in Flounder (Paralichthys olivaceus). [Master\u2019s Thesis, Northeast Agricultural University]."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Cheng, J., Yang, F., Liu, S., Zhao, H., Lu, W., and Zhang, Q. (2022). Transcriptomic analysis reveals functional interaction of mRNA-lncRAN-miRNA in steroidogenesis and spermatogenesis of gynogenetic Japanese flounder (Paralichthys olivceus). Biology, 11.","DOI":"10.3390\/biology11020213"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/11\/5935\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:50:09Z","timestamp":1760107809000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/11\/5935"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,29]]},"references-count":51,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2024,6]]}},"alternative-id":["ijms25115935"],"URL":"https:\/\/doi.org\/10.3390\/ijms25115935","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,29]]}}}