Expression and localization of PIWI proteins in testis and ovary of domestic sheep C., Liu Q., Wang X., Hu W., Han D., Mwacharo J.M., Wei C., Chu M., Di R. (2020): Expression and localization of PIWI proteins in testis and ovary of domestic sheep. Czech J. Anim. Sci., 65: 86-96.
download PDF

The piRNA-PIWI protein complex plays crucial roles in safeguarding the genome against inordinate transposon mobilization and regulation of embryonic development. A previous study indicated the presence of piRNA in sheep reproductive organs. However, the tissue distribution and cellular localization of PIWI proteins in sheep remains unclear. Therefore the present study aimed to explore the expression profiles of mRNAs of mammalian PIWI proteins (PIWIL1, PIWIL2, PIWIL4 and AGO3) in 9 tissues derived from adult male and female sheep. Results showed the expression of PIWIL1, PIWIL2, and PIWIL4 was significantly higher in the testis and ovary than in the other tissues. Immunohistochemistry analysis of testes indicated that each of the 4 proteins had specific cellular localizations, and some of the localizations were different from those of other species. All the proteins were mainly localized in the primary spermatocytes, suggesting that they are crucial for silencing of transposon to guarantee the integrity of the gamete genome during early stage of spermatogenesis. In the ovaries, the PIWI proteins were mainly localized in oocytes from antral follicles and leukocytes in ovarian blood. Our results provide insights to better understand the functions of PIWI proteins during spermatogenesis, oogenesis and immune defence in sheep.

Alazem M, He MH, Moffett P, Lin NS. Abscisic acid induces resistance against bamboo mosaic virus through Argonaute 2 and 3. Plant Physiol. 2017 May 1;174(1):339-55.
Aravin AA, Sachidanandam R, Bourc’his D, Schaefer C, Pezic D, Toth KF, Bestor T, Hannon GJ. A piRNA pathway primed by individual transposons is linked to de novo DNA methylation in mice. Mol Cell. 2008 Sep 26;31(6):785-99.
Batista PJ, Ruby JG, Claycomb JM, Chiang R, Fahlgren N, Kasschau KD, Chaves DA, Gu W, Vasale JJ, Duan S, Conte Jr D. PRG-1 and 21U-RNAs interact to form the piRNA complex required for fertility in C. elegans. Mol Cell. 2008 Jul 11;31(1):67-78.
Bortvin A. PIWI-interacting RNAs (piRNAs) − a mouse testis perspective. Biochemistry (Mosc). 2013 Jun 1;78(6):592-602.
Brennecke J, Aravin AA, Stark A, Dus M, Kellis M, Sachidanandam R, Hannon GJ. Discrete small RNA-generating loci as master regulators of transposon activity in Drosophila. Cell. 2007 Mar 23;128(6):1089-103.
Deng W, Lin H. Miwi, a murine homolog of piwi, encodes a cytoplasmic protein essential for spermatogenesis. Dev Cell. 2002 Jun 1;2(6):819-30.
Di R, He J, Song S, Tian D, Liu Q, Liang X, Ma Q, Sun M, Wang J, Zhao W, Cao G. Characterization and comparative profiling of ovarian microRNAs during ovine anestrus and the breeding season. BMC Genomics. 2014 Dec 1;15(1):899.
Ding X, Guan H, Li H. Characterization of a piRNA binding protein Miwi in mouse oocytes. Theriogenology. 2013 Mar 1;79(4):610-5.
Fang X, Chen C, Cai J, Xiang E, Li J, Chen P. Genome-wide methylation study of whole blood cells DNA in men with congenital hypopituitarism disease. Int J Mol Med. 2019 Jan 1;43(1):155-66.
Freedman JE, Gerstein M, Mick E, Rozowsky J, Levy D, Kitchen R, Das S, Shah R, Danielson K, Beaulieu L, Navarro FC. Diverse human extracellular RNAs are widely detected in human plasma. Nat Commun. 2016 Apr 26;7:11106.
Gainetdinov I, Skvortsova Y, Kondratieva S, Funikov S, Azhikina T. Two modes of targeting transposable elements by piRNA pathway in human testis. RNA. 2017 Nov 1;23(11):1614-25.
Goh WS, Falciatori I, Tam OH, Burgess R, Meikar O, Kotaja N, Hammell M, Hannon GJ. piRNA-directed cleavage of meiotic transcripts regulates spermatogenesis. Genes Dev. 2015 May 15;29(10):1032-44.
Gomes Fernandes M, He N, Wang F, Van Iperen L, Eguizabal C, Matorras R, Roelen BA, Chuva De Sousa Lopes SM. Human-specific subcellular compartmentalization of P-element induced wimpy testis-like (PIWIL) granules during germ cell development and spermatogenesis. Hum Reprod. 2018 Feb 1;33(2):258-69.
Gong J, Zhang Q, Wang Q, Ma Y, Du J, Zhang Y, Zhao X. Identification and verification of potential piRNAs from domesticated yak testis. Reproduction. 2018 Feb 1;155(2):117-27.
Gunawardane LS, Saito K, Nishida KM, Miyoshi K, Kawamura Y, Nagami T, Siomi H, Siomi MC. A slicer-mediated mechanism for repeat-associated siRNA 5’end formation in Drosophila. Science. 2007 Mar 16;315(5818):1587-90.
Hadziselimovic F, Hadziselimovic NO, Demougin P, Krey G, Oakeley E. Piwi-pathway alteration induces LINE-1 transposon derepression and infertility development in cryptorchidism. Sex Dev. 2015;9(2):98-104.
Han BW, Wang W, Li C, Weng Z, Zamore PD. piRNA-guided transposon cleavage initiates Zucchini-dependent, phased piRNA production. Science. 2015 May 15;348(6236):817-21.
Han YN, Li Y, Xia SQ, Zhang YY, Zheng JH, Li W. PIWI proteins and PIWI-interacting RNA: emerging roles in cancer. Cell Physiol Biochem. 2017;44(1):1-20.
Hempfling AL, Lim SL, Adelson DL, Evans J, O’Connor AE, Qu ZP, Kliesch S, Weidner W, O’Bryan MK, Bergmann M. Expression patterns of HENMT1 and PIWIL1 in human testis: implications for transposon expression. Reproduction. 2017 Oct 1;154(4):363-74.
Hirano T, Iwasaki YW, Lin ZY, Imamura M, Seki NM, Sasaki E, Saito K, Okano H, Siomi MC, Siomi H. Small RNA profiling and characterization of piRNA clusters in the adult testes of the common marmoset, a model primate. RNA. 2014 Aug 1;20(8):1223-37.
Houwing S, Kamminga LM, Berezikov E, Cronembold D, Girard A, Van Den Elst H, Filippov DV, Blaser H, Raz E, Moens CB, Plasterk RH. A role for Piwi and piRNAs in germ cell maintenance and transposon silencing in Zebrafish. Cell. 2007 Apr 6;129(1):69-82.
Huang RC, Garratt ES, Pan H, Wu Y, Davis EA, Barton SJ, Burdge GC, Godfrey KM, Holbrook JD, Lillycrop KA. Genome-wide methylation analysis identifies differentially methylated CpG loci associated with severe obesity in childhood. Epigenetics. 2015 Nov 2;10(11):995-1005.
Kabayama Y, Toh H, Katanaya A, Sakurai T, Chuma S, Kuramochi-Miyagawa S, Saga Y, Nakano T, Sasaki H. Roles of MIWI, MILI and PLD6 in small RNA regulation in mouse growing oocytes. Nucleic Acids Res. 2017 May 19;45(9):5387-98.
Kang HJ, Moon MJ, Lee HY, Han SW. Testosterone alters testis function through regulation of piRNA expression in rats. Mol Biol Rep. 2014 Oct 1;41(10):6729-35.
Kimura M, Ishida K, Kashiwabara SI, Baba T. Characterization of two cytoplasmic poly (A)-binding proteins, PABPC1 and PABPC2, in mouse spermatogenic cells. Biol Reprod. 2009 Mar 1;80(3):545-54.
Kirino Y, Kim N, de Planell-Saguer M, Khandros E, Chiorean S, Klein PS, Rigoutsos I, Jongens TA, Mourelatos Z. Arginine methylation of Piwi proteins catalysed by dPRMT5 is required for Ago3 and Aub stability. Nat Cell Biol. 2009 May;11(5):652-8.
Kowalczykiewicz D, Pawlak P, Lechniak D, Wrzesinski J. Altered expression of porcine Piwi genes and piRNA during development. PLoS One. 2012;7(8):e43816.
Krawczynski K, Najmula J, Bauersachs S, Kaczmarek MM. MicroRNAome of porcine conceptuses and trophoblasts: expression profile of micrornas and their potential to regulate genes crucial for establishment of pregnancy. Biol Reprod. 2015 Jan 1;92(1):1-22.
Lee JH, Schutte D, Wulf G, Fuzesi L, Radzun HJ, Schweyer S, Engel W, Nayernia K. Stem-cell protein Piwil2 is widely expressed in tumors and inhibits apoptosis through activation of Stat3/Bcl-XL pathway. Hum Mol Genet. 2006 Jan 15;15(2):201-11.
Lim SL, Tsend-Ayush E, Kortschak RD, Jacob R, Ricciardelli C, Oehler MK, Grutzner F. Conservation and expression of PIWI-interacting RNA pathway genes in male and female adult gonad of amniotes. Biol Reprod. 2013 Dec 1;89(6):136.
Lin H, Yin H. A novel epigenetic mechanism in Drosophila somatic cells mediated by Piwi and piRNAs. Cold Spring Harb Symp Quant Biol. 2008 Jan 1;73(1):273-81.
Luteijn MJ, Ketting RF. PIWI-interacting RNAs: from generation to transgenerational epigenetics. Nat Rev Genet. 2013 Aug;14(8):523-34.
Meister G. Argonaute proteins: Functional insights and emerging roles. Nat Rev Genet. 2013 Jul;14(7):447-59.
Miesen P, Girardi E, van Rij RP. Distinct sets of PIWI proteins produce arbovirus and transposon-derived piRNAs in Aedes aegypti mosquito cells. Nucleic Acids Res. 2015 Jul 27;43(13):6545-56.
Nishida KM, Saito K, Mori T, Kawamura Y, Nagami-Okada T, Inagaki S, Siomi H, Siomi MC. Gene silencing mechanisms mediated by Aubergine–piRNA complexes in Drosophila male gonad. RNA. 2007 Nov 1;13(11):1911-22.
Parikh RY, Lin H, Gangaraju VK. A critical role for nucleoporin 358 (Nup358) in transposon silencing and piRNA biogenesis in Drosophila. J Biol Chem. 2018 Jun 15;293(24):9140-7.
Perera BP, Tsai ZT, Colwell ML, Jones TR, Goodrich JM, Wang K, Sartor MA, Faulk C, Dolinoy DC. Somatic expression of piRNA and associated machinery in the mouse identifies short, tissue-specific piRNA. Epigenetics. 2019 May 4;14(5):504-21.
Pillai RS, Chuma S. piRNAs and their involvement in male germline development in mice. Dev Growth Differ. 2012 Jan;54(1):78-92.
Russell SJ, Stalker L, Gilchrist G, Backx A, Molledo G, Foster RA, LaMarre J. Identification of PIWIL1 isoforms and their expression in bovine testes, oocytes, and early embryos. Biol Reprod. 2016 Apr 1;94(4):75.
Saint-Leandre B, Clavereau I, Hua-Van A, Capy P. Transcriptional polymorphism of pi RNA regulatory genes underlies the mariner activity in Drosophila simulans testes. Mol Ecol. 2017 Jul;26(14):3715-31.
Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008 Jun;3(6):1101-8.
Siomi MC, Sato K, Pezic D, Aravin AA. PIWI-interacting small RNAs: The vanguard of genome defence. Nat Rev Mol Cell Biol. 2011 Apr;12(4):246-58.
Sugimoto K, Kage H, Aki N, Sano A, Kitagawa H, Nagase T, Yatomi Y, Ohishi N, Takai D. The induction of H3K9 methylation by PIWIL4 at the p16Ink4a locus. Biochem Biophys Res Commun. 2007 Aug 3;359(3):497-502.
Thomson T, Lin H. The biogenesis and function of PIWI proteins and piRNAs: progress and prospect. Annu Rev Cell Dev Biol. 2009 Nov 10;25(1):355-76.
Tosar JP, Rovira C, Cayota A. Non-coding RNA fragments account for the majority of annotated piRNAs expressed in somatic non-gonadal tissues. Commun Biol. 2018 Jan 22;1(1):1-8.
Toth KF, Pezic D, Stuwe E, Webster A. The piRNA pathway guards the germline genome against transposable elements. Adv Exp Med Biol. 2016 Dec;886:51-77.
Untergasser A, Cutcutache I, Koressaar T, Ye J, Faircloth BC, Remm M, Rozen SG. Primer3 − new capabilities and interfaces. Nucleic Acids Res. 2012 Aug 1;40(15):e115.
Van Stry M, Oguin TH, Cheloufi S, Vogel P, Watanabe M, Pillai MR, Dash P, Thomas PG, Hannon GJ, Bix M. Enhanced susceptibility of Ago1/3 double-null mice to influenza A virus infection. J Virol. 2012 Apr 15;86(8):4151-7.
Varjak M, Maringer K, Watson M, Sreenu VB, Fredericks AC, Pondeville E, Donald CL, Sterk J, Kean J, Vazeille M, Failloux AB. Aedes aegypti Piwi4 is a noncanonical PIWI protein involved in antiviral responses. MSphere. 2017a Jun 28;2(3):1-16.
Varjak M, Donald CL, Mottram TJ, Sreenu VB, Merits A, Maringer K, Schnettler E, Kohl A. Characterization of the Zika virus induced small RNA response in Aedes aegypti cells. PLoS Negl Trop Dis. 2017b Oct 17;11(10):e0006010.
Varjak M, Dietrich I, Sreenu VB, Till BE, Merits A, Kohl A, Schnettler E. Spindle-e acts antivirally against alphaviruses in mosquito cells. Viruses. 2018 Feb;10(2):88.
Wang D, Zhang Z, O’Loughlin E, Lee T, Houel S, O’Carroll D, Tarakhovsky A, Ahn NG, Yi R. Quantitative functions of Argonaute proteins in mammalian development. Genes Dev. 2012 Apr 1;26(7):693-704.
Wang W, Yoshikawa M, Han BW, Izumi N, Tomari Y, Weng Z, Zamore PD. The initial uridine of primary piRNAs does not create the tenth adenine that Is the hallmark of secondary piRNAs. Mol Cell. 2014 Dec 4;56(5):708-16.
Wang HZ, Ma T, Chang GB, Wan F, Liu XP, Lu L, Xu L, Chen J, Chen GH. Single nucleotide polymorphism screening, molecular characterization, and evolutionary aspects of chicken Piwi genes. Genet Mol Res. 2015 Jan 1;14(4):14802-10.
Wang H, Wang B, Liu X, Liu Y, Du X, Zhang Q, Wang X. Identification and expression of piwil2 in turbot Scophthalmus maximus, with implications of the involvement in embryonic and gonadal development. Comp Biochem Physiol B Biochem Mol Biol. 2017 Jun 1;208-209:84-93.
Wang H, Wang B, Liu J, Li A, Zhu H, Wang X, Zhang Q. Piwil1 gene is regulated by hypothalamic-pituitary-gonadal axis in turbot (Scophthalmus maximus): A different effect in ovaries and testes. Gene. 2018 Jun 5;658:86-95.
Wu Y, Xu K, Qi H. Domain-functional analyses of PIWIL1 and PABPC1 indicate their synergistic roles in protein translation via 3'-UTRs of meiotic mRNAs. Biol Reprod. 2018 Oct 1;99(4):773-88.
Xie K, Zhang K, Kong J, Wang C, Gu Y, Liang C, Jiang T, Qin N, Liu J, Guo X, Huo R. Cancer-testis gene PIWIL 1 promotes cell proliferation, migration, and invasion in lung adenocarcinoma. Cancer Med. 2018 Jan;7(1):157-66.
Xu K, Yang L, Zhao D, Wu Y, Qi H. AKAP3 synthesis is mediated by RNA binding proteins and PKA signaling during mouse spermiogenesis. Biol Reprod. 2014 Jun 1;90(6):119.
Zhao SZ, Li Y, Jiang X, Lu YL, Tao DC, Liu YQ, Ma YX. Identification and expression analysis of Macaca mulatta piwil4 gene. Yi chuan. 2011 Apr;33(4):365-70.
Zhou X, Guo H, Chen K, Cheng H, Zhou R. Identification, chromosomal mapping and conserved synteny of porcine Argonaute family of genes. Genetica. 2010 Jul 1;138(7):805-12.
download PDF

© 2021 Czech Academy of Agricultural Sciences | Prohlášení o přístupnosti