FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Animals (Basel) / Comparative Transcriptomics Analysis of Testicular miRNA fro...
Animals : an open access journal from MDPI2020; 10(2); doi: 10.3390/ani10020338

Comparative Transcriptomics Analysis of Testicular miRNA from Cryptorchid and Normal Horses.

Abstract: In the biological process of testicular spermatogenesis, the expression and interaction of many genes are regulated by microRNAs (miRNAs). However, comparisons of miRNA expression between descended testes (DTs) and undescended testes (UDTs) are rarely done in horses. In this study, we selected two UDTs (CKY2b and GU4b) from Chakouyi (CKY) and Guanzhong (GU) horses and eight DTs (GU1-3, CKY1, CKY3, CKY2a, GU4a, and GU5). Three groups were compared to evaluate expression patterns of testicular miRNA in stallion testes. Group 1 compared normal CKY horses and GU horses (CKY1 and CKY3 vs. GU1-3). Group 2 (CKY2a and GU4a (DTs) vs. CKY2b and GU4b (UDTs)) and group 3 (GU1-3, CKY1, CKY3 (DTs) vs. CKY2b and GU4b (UDTs)) compared the expression levels in unilateral retained testes to normal testes. The results show that 42 miRNAs (7 upregulated and 35 downregulated) had significantly different expression levels in both comparisons. The expression levels of eca-miR-545, eca-miR-9084, eca-miR-449a, eca-miR-9024, eca-miR-9121, eca-miR-8908e, eca-miR-136, eca-miR-329b, eca-miR-370, and eca-miR-181b were further confirmed by quantitative real-time PCR assay. The target genes of differentially expressed miRNAs in three comparisons were predicted, and the functions were annotated. The putative target genes of the 42 co-differentially expressed miRNAs were annotated to 15 functional terms, including metal ion binding, GTPase activator activity, zinc ion binding, intracellular, cytoplasm, and cancer pathways, and osteoclast differentiation. Our data indicate that the differentially expressed miRNAs in undescended testis suggests a potential role in male fertility and a relationship with cryptorchidism in horses. The discovery of miRNAs in stallion testes might contribute to a new direction in the search for biomarkers of stallion fertility.
Get Full Text
Publication Date: 2020-02-21 PubMed ID: 32098036PubMed Central: PMC7070967DOI: 10.3390/ani10020338Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research study undertakes a comparison of gene regulation in the testes of normal horses and those with an undescended testes condition called cryptorchidism. The gene activity regulation is done by microRNAs (miRNAs), which are studied to understand their effect on male fertility.

Objective of the Study

  • This study aimed to examine and compare the microRNA (miRNA) expression between descended testes (DTs) and undescended testes (UDTs) in horses, a rarely done comparison.

Methodology Adopted

  • Testicular miRNA expression in stallion testes was evaluated in three comparison groups. Group 1 compared normal horses from the Chakouyi (CKY) and Guanzhong (GU) breeds, Group 2 compared the expression levels in normal testes versus unilateral retained testes, and Group 3 also compared normal testes with unilateral retained testes, but with a different selection.
  • The expression levels of specific miRNAs were confirmed through a quantitative real-time PCR assay.
  • The study also predicted the target genes of differentially expressed miRNAs and annotated their functions.

Key Findings of the Study

  • The research found that 42 miRNAs exhibited significantly different expression levels in both comparisons, with seven being upregulated and 35 being downregulated.
  • The putative target genes of co-differentially expressed miRNAs were linked to 15 functional terms. These included metal ion binding, GTPase activator activity, zinc ion binding, intracellular activity, cytoplasm, cancer pathways, and osteoclast differentiation.
  • These variations in expression levels imply a possible impact on male fertility and indicate a relationship with cryptorchidism in horses.

Conclusion and Implications of the Study

  • The results suggest that the expression of certain miRNAs in undescended testes may potentially influence male fertility in horses.
  • This discovery of miRNAs in stallion testes can contribute to exploring biomarkers of stallion fertility, thereby setting a new direction for future research in this area.

Cite This Article

APA
Han H, Chen Q, Gao Y, Li J, Li W, Dang R, Lei C. (2020). Comparative Transcriptomics Analysis of Testicular miRNA from Cryptorchid and Normal Horses. Animals (Basel), 10(2). https://doi.org/10.3390/ani10020338

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 10
Issue: 2

Researcher Affiliations

Han, Haoyuan
  • College of Animal Science and Technology, Henan University of Animal Husbandry and Economy, Zhengzhou, Henan 450046, China.
  • College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.
Chen, Qiuming
  • College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.
Gao, Yuan
  • College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.
Li, Jun
  • College of Animal Science and Technology, Henan University of Animal Husbandry and Economy, Zhengzhou, Henan 450046, China.
Li, Wantao
  • Henan Genetic Protection Engineering Research Center for Livestock and Poultry, Zhengzhou 450046, Henan, China.
Dang, Ruihua
  • College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.
Lei, Chuzhao
  • College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.

Grant Funding

  • 81270439 / National Natural Science Foundation of China
  • 906/24030042 and 906/24030103 / PhD Start-up Fund of Henan University of Animal Husbandry and Economy

Conflict of Interest Statement

The authors declare that they have no conflict of interest.

References

This article includes 47 references
  1. Bartel D.P.. MicroRNAs: Genomics, biogenesis, mechanism, and function.. Cell 2004;116:281–297.
    doi: 10.1016/S0092-8674(04)00045-5pubmed: 14744438google scholar: lookup
  2. Pritchard C., Cheng H., Tewari M.. MicroRNA profiling: Approaches and considerations.. Nat. Rev. Genet. 2012;13:358–369.
    doi: 10.1038/nrg3198pmc: PMC4517822pubmed: 22510765google scholar: lookup
  3. Lagos-Quintana M., Rauhut R., Meyer J., Borkhardt A., Tuschl T.. New microRNAs from mouse and human.. RNA 2003;9:175–179.
    doi: 10.1261/rna.2146903pmc: PMC1370382pubmed: 12554859google scholar: lookup
  4. Lagos-Quintana M., Rauhut R., Lendeckel W., Tuschl T.. Identification of Novel Genes Coding for Small Expressed RNAs.. Science 2001;294:853–858.
    doi: 10.1126/science.1064921pubmed: 11679670google scholar: lookup
  5. Gregory R.I., Yan K.P., Amuthan G., Chendrimada T., Doratotaj B., Cooch N.. The Microprocessor complex mediates the genesis of microRNAs.. Nature 2004;432:235–240.
    doi: 10.1038/nature03120pubmed: 15531877google scholar: lookup
  6. Chen C.Z., Li L., Lodish H.F., Bartel D.P.. MicroRNAs modulate hematopoietic lineage differentiation.. Science 2004;303:83–86.
    doi: 10.1126/science.1091903pubmed: 14657504google scholar: lookup
  7. Kloosterman W.P., Plasterk R.H.. The diverse functions of microRNAs in animal development and disease.. Dev. Cell 2006;11:441–450.
    doi: 10.1016/j.devcel.2006.09.009pubmed: 17011485google scholar: lookup
  8. Stefani G., Slack F.J.. Small non-coding RNAs in animal development.. Nat. Rev. Mol. Cell Biol. 2008;9:219–230.
    doi: 10.1038/nrm2347pubmed: 18270516google scholar: lookup
  9. Mendell J.T., Olson E.N.. MicroRNAs in stress signaling and human disease.. Cell 2012;148:1172–1187.
    doi: 10.1016/j.cell.2012.02.005pmc: PMC3308137pubmed: 22424228google scholar: lookup
  10. Maatouk D.M., Loveland K.L., McManus M.T.. Dicer1 is required for differentiation of the mouse male germline.. Biol. Reprod. 2008;79:696–703.
    doi: 10.1095/biolreprod.108.067827pubmed: 18633141google scholar: lookup
  11. Huff D.S., Fenig D.M., Canning D.A., Carr M.G., Zderic S.A., Snyder H.M.. Abnormal germ cell development in cryptorchidism.. Horm. Res. 2001;55:11–17.
    doi: 10.1159/000049957pubmed: 11423736google scholar: lookup
  12. Nguyen M.T., Delaney D.P., Kolon T.F.. Gene expression alterations in cryptorchid males using spermatozoal microarray analysis.. Fertil. Steril. 2009;92:182–187.
    doi: 10.1016/j.fertnstert.2008.05.043pubmed: 18687424google scholar: lookup
  13. Hadziselimovic F., Herzog B.. The importance of both an early orchidopexy and germ cell maturation for fertility.. Lancet 2001;358:1156–1157.
    doi: 10.1016/S0140-6736(01)06274-2pubmed: 11597673google scholar: lookup
  14. Carreau S., Lambard S., Said L., Saad A., Galeraud-Denis I.. RNA dynamics of fertile and infertile spermatozoa.. Biochem. Soc. Trans. 2007;35:634–636.
    doi: 10.1042/BST0350634pubmed: 17511668google scholar: lookup
  15. Platts A.E., Dix D.J., Chemes H.E., Thompson K.E., Goodrich R., Rockett J.C., Rawe V.Y., Quintana S., Diamond M.P., Strader L.F.. Success and failure in human spermatogenesis as revealed by teratozoospermic RNAs.. Hum. Mol. Genet. 2007;16:763–773.
    doi: 10.1093/hmg/ddm012pubmed: 17327269google scholar: lookup
  16. Garrido N., Martínez-Conejero J.A., Jauregui J., Horcajadas J.A., Simón C., Remohí J., Meseguer M.. Microarray analysis in sperm from fertile and infertile men without basic sperm analysis abnormalities reveals a significantly different transcriptome.. Fertil. Steril. 2009;91:1307–1310.
    doi: 10.1016/j.fertnstert.2008.01.078pubmed: 18367176google scholar: lookup
  17. Bissonnette N., Levesque-Sergerie J.P., Thibault C., Boissonneault G.. Spermatozoal transcriptome profiling for bull sperm motility: A potential tool to evaluate semen quality.. Reproduction 2009;138:65–80.
    doi: 10.1530/REP-08-0503pubmed: 19423662google scholar: lookup
  18. Tong M.H., Mitchell D.A., McGowan S.D., Evanoff R., Griswold M.D.. Two miRNA clusters, Mir-17-92 (Mirc1) and Mir-106b-25(Mirc3), are involved in the regulation of spermatogonial differentiation in mice.. Biol. Reprod. 2012;86:72.
    doi: 10.1095/biolreprod.111.096313pmc: PMC3316268pubmed: 22116806google scholar: lookup
  19. Yang C.C., Lin Y.S., Hsu C.C., Wu S.C., Lin E.C., Cheng W.T.. Identification and sequencing of remnant messenger RNAs found in domestic swine (Sus scrofa) fresh ejaculated spermatozoa.. Anim. Reprod. Sci. 2009;113:143–155.
    doi: 10.1016/j.anireprosci.2008.08.012pubmed: 18786788google scholar: lookup
  20. Yu Z., Raabe T., Hecht N.B.. MicroRNA Mirn122a reduces expression of the posttranscriptionally regulated germ cell transition protein 2 (Tnp2) messenger RNA (mRNA) by mRNA cleavage.. Biol. Reprod. 2005;73:427–433.
    doi: 10.1095/biolreprod.105.040998pubmed: 15901636google scholar: lookup
  21. Moritoki Y., Hayashi Y., Mizuno K., Kamisawa H., Nishio H., Kurokawa S., Ugawa S., Kojima Y., Kohri K.. Expression profiling of microRNA in cryptorchid testes: miR-135a contributes to the maintenance of spermatogonial stem cells by regulating FoxO1.. J. Urol. 2014;191:1174–1180.
    doi: 10.1016/j.juro.2013.10.137pubmed: 24184258google scholar: lookup
  22. Das P.J., Fiona M.C., Monika V., Nandina P., Cathy G., Gang L., Priyanka K., Sudderth A.K., Teague S., Love C.C.. Stallion Sperm Transcriptome Comprises Functionally Coherent Coding and Regulatory RNAs as Revealed by Microarray Analysis and RNA-seq.. PLoS ONE 2013;8:e56535.
    doi: 10.1371/journal.pone.0056535pmc: PMC3569414pubmed: 23409192google scholar: lookup
  23. Mortazavi A., Williams B.A., McCue K., Schaeffer L., Wold B.. Mapping and quantifying mammalian transcriptomes by RNA-Seq.. Nat. Methods 2008;5:621–628.
    doi: 10.1038/nmeth.1226pubmed: 18516045google scholar: lookup
  24. Urh K., Kunej T.. Molecular mechanisms of cryptorchidism development: Update of the database, disease comorbidity, and initiative for standardization of reporting in scientific literature.. Andrology 2016;4:894–902.
    doi: 10.1111/andr.12217pubmed: 27370962google scholar: lookup
  25. Paria N., Raudsepp T., Pearks Wilkerson A.J., O’Brien P.C., Ferguson-Smith M.A., Love C.C., Arnold C., Rakestraw P., Murphy W.J., Chowdhary B.P.. A gene catalogue of the euchromatic male-specific region of the horse Y chromosome: Comparison with human and other mammals.. PLoS ONE 2011;6:e21374.
    doi: 10.1371/journal.pone.0021374pmc: PMC3143126pubmed: 21799735google scholar: lookup
  26. Love M.I., Huber W., Anders S.. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2.. Genome Biol. 2014;15:550.
    doi: 10.1186/s13059-014-0550-8pmc: PMC4302049pubmed: 25516281google scholar: lookup
  27. Schmittgen T.D., Qian L., Yang L.. Diverse expression of microRNA precursors in human cancer cell lines.. Cancer Res. 2004;45:64.
  28. Vasileva A., Tiedau D., Firooznia A., Müller-Reichert T., Jessberger R.. Tdrd6 is required for spermiogenesis, chromatoid body architecture, and regulation of miRNA expression.. Curr. Biol. 2009;19:630–639.
    doi: 10.1016/j.cub.2009.02.047pmc: PMC2719840pubmed: 19345099google scholar: lookup
  29. Buchold G.M., Coarfa C., Kim J., Milosavljevic A., Gunaratne P.H., Matzuk M.M.. Analysis of microRNA expression in the prepubertal testis.. PLoS ONE 2010;5:e15317.
    doi: 10.1371/journal.pone.0015317pmc: PMC3012074pubmed: 21206922google scholar: lookup
  30. Niu Z., Goodyear S., Rao S., Wu X., Tobias J.W., Avarbock M.R., Brinster R.L.. MicroRNA-21 regulates the self-renewal of mouse spermatogonial stem cells.. PNAS 2011;108:12740–12745.
    doi: 10.1073/pnas.1109987108pmc: PMC3150879pubmed: 21768389google scholar: lookup
  31. Dai L., Tsai-Morris C.H., Sato H., Villar J., Kang J.H., Zhang J.M.L.. Testis-specific miRNA-469 up-regulated in gonad otropinregulated testicular RNA helicase (GRTH/DDX25)-null mice silences transition protein 2 and protamine 2 messages at sites within coding region. Implications of its role in germ cell development.. J. Biol. Chem. 2011;286:52.
    doi: 10.1074/jbc.M111.282756pmc: PMC3248001pubmed: 22086916google scholar: lookup
  32. Mclver S.C., Stanger S.J., Santarelli D.M., Roman S.D., Nixon B., McLaughlin E.A.. A unique combination of male germ cell miRNAs coordinates gonocyte differentiation.. PLoS ONE 2012;7:e35553.
    pmc: PMC3334999pubmed: 22536405
  33. Hayashi T., Kageyama Y., Ishizaka K., Xia G., Kihara K., Oshima H.. Requirement of Notch 1 and its ligand jagged 2 expressions for spermatogenesis in rat and human testes.. J. Androl. 2001;22:999–1011.
    doi: 10.1002/j.1939-4640.2001.tb03441.xpubmed: 11700865google scholar: lookup
  34. Liu W.M., Pang R.T.K., Chiu P.C.N., Wong B.P.C., Lao K., Lee K.F., Yeung W.S.B.. Sperm-borne microRNA-34c is required for the first cleavage division in mouse.. PNAS 2012;106:490–494.
    doi: 10.1073/pnas.1110368109pmc: PMC3258645pubmed: 22203953google scholar: lookup
  35. Yamamoto C.M., Hikim A.P., Lue Y., Portugal A.M., Guo T.B., Hsu S.Y., Salameh W.A., Wang C., Hsueh A.J., Swerdloff R.S.. Impairment of spermatogenesis in transgenic mice with selective overexpression of Bcl-2 in the somatic cells of the testis.. J. Androl. 2001;22:981–991.
    doi: 10.1002/j.1939-4640.2001.tb03439.xpubmed: 11700863google scholar: lookup
  36. Selbach M., Schwanhäusser B., Thierfelder N., Fang Z., Khanin R., Rajewsky N.. Widespread changes in protein synthesis induced by microRNAs.. Nature 2008;7209:58–63.
    doi: 10.1038/nature07228pubmed: 18668040google scholar: lookup
  37. Yan N., Lu Y., Sun H., Qiu W., Tao D., Liu Y., Chen H., Yang Y., Zhang S., Li X.. Microarray profiling of microRNAs expressed in testis tissues of developing primates.. J. Assist. Reprod. Genet. 2009;26:179–186.
    doi: 10.1007/s10815-009-9305-ypmc: PMC2682186pubmed: 19242788google scholar: lookup
  38. Yang G., Zhang Y.L., Buchold G.M., Jetten A.M., O’Brien D.A.. Analysis of germ cell nuclear factor transcripts and protein expression during spermatogenesis.. Biol. Reprod. 2003;68:1620–1630.
    doi: 10.1095/biolreprod.102.012013pubmed: 12606326google scholar: lookup
  39. Connor F., Wright E., Denny P., Koopman P., Ashworth A.. The Sryrelated HMG box-containing gene Sox6 is expressed in the adult testis and developing nervous system of the mouse.. Nucleic Acids Res. 1995;23:3365–3372.
    doi: 10.1093/nar/23.17.3365pmc: PMC307212pubmed: 7567444google scholar: lookup
  40. Serge C., Rex A.H.. Oestrogens and spermatogenesis.. Philos. Trans. R. Soc. Lond. B Biol. Sci. 2010;365:1517–1535.
    pmc: PMC2871919pubmed: 20403867
  41. Hannema S.E., Scott I.S., Rajpert De Meyts E., Skakkebaek N.E., Coleman N., Hughes I.A.. Testicular development in the complete androgen insensitivity syndrome.. J. Pathol. 2010;208:518–527.
    doi: 10.1002/path.1890pubmed: 16400621google scholar: lookup
  42. Vandenput L., Ederveen A., Erben R., Stahr K., Swinnen J.V., Herch E.V., Verstuyf A., Boonen S., Bouillon R., Vanderschueren D.. Testosterone prevents orchidectomy-induced bone loss in estrogen receptor-alpha knockout mice.. Biochem. Biophys. Res. Commun. 2001;285:70–76.
    doi: 10.1006/bbrc.2001.5101pubmed: 11437374google scholar: lookup
  43. Franzoso G.. Requirement for NF-κB in osteoclast and B-cell development.. Genes Dev. 1997;11:3482–3496.
    doi: 10.1101/gad.11.24.3482pmc: PMC316809pubmed: 9407039google scholar: lookup
  44. Girard A., Sachidanandam R., Hannon G.J., Carmell M.A.. A germlinespecific class of small RNAs binds mammalian Piwi proteins.. Nature 2006;442:199–202.
    doi: 10.1038/nature04917pubmed: 16751776google scholar: lookup
  45. Kawano M., Kawaji H., Grandjean V., Kiani J., Rassoulzadegan M.. Novel small noncoding RNAs in mouse spermatozoa, zygotes and early embryos.. PLoS ONE 2012;7:e44542.
    doi: 10.1371/journal.pone.0044542pmc: PMC3440372pubmed: 22984523google scholar: lookup
  46. Sai Lakshmi S., Agrawal S.. piRNABank: A web resource on classified and clustered Piwi-interacting RNAs.. Nucleic Acids Res. 2008;36:D173–D177.
    doi: 10.1093/nar/gkm696pmc: PMC2238943pubmed: 17881367google scholar: lookup
  47. Thomson T., Lin H.. The biogenesis and function of PIWI proteins and piRNAs: Progress and prospect.. Annu. Rev. Cell Dev. Biol. 2009;25:355–376.
    doi: 10.1146/annurev.cellbio.24.110707.175327pmc: PMC2780330pubmed: 19575643google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.