{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T21:06:57Z","timestamp":1789420017053,"version":"build-2803163510"},"reference-count":33,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2021,10,20]],"date-time":"2021-10-20T00:00:00Z","timestamp":1634688000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The four human PIWI-LIKE gene family members PIWI-LIKE 1\u20134 play a pivotal role in stem cell maintenance and transposon repression in the human germline. Therefore, dysregulation of these genes negatively influences the genetic stability of the respective germ cell and subsequent development and maturation. Recently, we demonstrated that a lower PIWI-LIKE 2 mRNA expression in ejaculated spermatozoa is more frequent in men with oligozoospermia. In this study, we analysed how PIWI-LIKE 1\u20134 mRNA expression in ejaculated spermatozoa predicts ART outcome. From 160 IVF or ICSI cycles, portions of swim-up spermatozoa used for fertilization were collected, and the total RNA was isolated. PIWI-LIKE 1\u20134 mRNA expression was measured by qPCR using TaqMan probes with GAPDH as a reference gene. PIWI-LIKE 1 and 2 transcript levels in the spermatozoa of the swim-up fraction were positively correlated to each other (rS = 0.78; p &lt; 0.001). Moreover, lower PIWI-LIKE 2 mRNA levels, as well as lower PIWI-LIKE 1 mRNA levels, in these spermatozoa were positively associated with a fertilization rate \u2265 50% in the respective ART cycles (p = 0.02 and p = 0.0499, Mann\u2013Whitney U-Test). When separately analysing IVF and ICSI cycles, PIWI-LIKE 1 and 2 transcript levels were only significantly associated to increased fertilization rates in IVF, yet not in ICSI cycles. Spermatozoal PIWI-LIKE 3 and 4 transcript levels were not significantly associated to fertilization rates in ART cycles. In conclusion, lower levels of spermatozoal PIWI-LIKE 1 and 2 mRNA levels are positively associated with a higher fertilization rate in IVF cycles.<\/jats:p>","DOI":"10.3390\/ijms222111320","type":"journal-article","created":{"date-parts":[[2021,10,20]],"date-time":"2021-10-20T21:29:46Z","timestamp":1634765386000},"page":"11320","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Lower Spermatozoal PIWI-LIKE 1 and 2 Transcript Levels Are Significantly Associated with Higher Fertilization Rates in IVF"],"prefix":"10.3390","volume":"22","author":[{"given":"Maria","family":"Giebler","sequence":"first","affiliation":[{"name":"Center for Reproductive Medicine and Andrology, Martin Luther University Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1888-7813","authenticated-orcid":false,"given":"Thomas","family":"Greither","sequence":"additional","affiliation":[{"name":"Center for Reproductive Medicine and Andrology, Martin Luther University Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diana","family":"Handke","sequence":"additional","affiliation":[{"name":"Center for Reproductive Medicine and Andrology, Martin Luther University Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gregor","family":"Seliger","sequence":"additional","affiliation":[{"name":"Center for Reproductive Medicine and Andrology, Martin Luther University Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hermann M.","family":"Behre","sequence":"additional","affiliation":[{"name":"Center for Reproductive Medicine and Andrology, Martin Luther University Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1186\/s12958-015-0032-1","article-title":"A unique view on male infertility around the globe","volume":"13","author":"Agarwal","year":"2015","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1093\/aje\/kwp168","article-title":"Good Semen Quality and Life Expectancy: A Cohort Study of 43,277 Men","volume":"170","author":"Jensen","year":"2009","journal-title":"Am. J. Epidemiol."},{"key":"ref_3","first-page":"2266","article-title":"Male factor infertility and risk of death: A nationwide record-linkage study","volume":"34","author":"Glazer","year":"2019","journal-title":"Hum. Reprod."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"313","DOI":"10.3109\/19396368.2012.706361","article-title":"Segregation of sperm subpopulations in normozoospermic infertile men","volume":"58","author":"Thilagavathi","year":"2012","journal-title":"Syst. Biol. Reprod. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2749","DOI":"10.18632\/aging.101946","article-title":"Male age: Negative impact on sperm DNA fragmentation","volume":"11","author":"Albani","year":"2019","journal-title":"Aging"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Poenicke, K., Grunewald, S., Glander, H., Paasch, U., Kamini, A.R., Agarwal, A., and Srinivas, M. (2010). Sperm Selection in Assisted Reproductive Techniques. Andrology Laboratory Manual, Jaypee Brothers Pvt Ltd.","DOI":"10.5005\/jp\/books\/11172_18"},{"key":"ref_7","first-page":"e13718","article-title":"Comparative analysis of tests used to assess sperm chromatin integrity and DNA fragmentation","volume":"53","author":"Dutta","year":"2020","journal-title":"Andrologia"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1016\/j.theriogenology.2005.09.011","article-title":"Clinical aspects of sperm DNA fragmentation detection and male infertility","volume":"65","author":"Evenson","year":"2006","journal-title":"Theriogenology"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"101868","DOI":"10.1016\/j.jogoh.2020.101868","article-title":"Predictive value of the sperm DNA fragmentation index for low or failed IVF fertilization in men with mild-to-moderate asthenozoospermia","volume":"50","author":"Tang","year":"2020","journal-title":"J. Gynecol. Obstet. Hum. Reprod."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1111\/andr.12153","article-title":"Sperm chromatin structure assay in prediction of in vitro fertilization outcome","volume":"4","author":"Oleszczuk","year":"2016","journal-title":"Andrology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"352","DOI":"10.1016\/j.fertnstert.2007.06.018","article-title":"Sperm chromatin structure assay parameters are not related to fertilization rates, embryo quality, and pregnancy rates in in vitro fertilization and intracytoplasmic sperm injection, but might be related to spontaneous abortion rates","volume":"90","author":"Lin","year":"2008","journal-title":"Fertil. Steril."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1007\/s10815-019-01632-5","article-title":"Sperm DNA fragmentation on the day of fertilization is not associated with embryologic or clinical outcomes after IVF\/ICSI","volume":"37","author":"Green","year":"2019","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1609","DOI":"10.1093\/humrep\/deq116","article-title":"Fall in implantation rates following ICSI with sperm with high DNA fragmentation","volume":"25","author":"Speyer","year":"2010","journal-title":"Hum. Reprod."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e13263","DOI":"10.1111\/and.13263","article-title":"Sperm DNA fragmentation index influences assisted reproductive technology outcome: A systematic review and meta-analysis combined with a retrospective cohort study","volume":"51","author":"Deng","year":"2019","journal-title":"Andrologia"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"246","DOI":"10.4103\/aja.aja_51_19","article-title":"Implementation of an in-house flow cytometric analysis of DNA fragmentation in spermatozoa","volume":"22","author":"Rex","year":"2020","journal-title":"Asian J. Androl."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"39","DOI":"10.6061\/clinics\/2013(Sup01)06","article-title":"A comprehensive review of genetics and genetic testing in azoospermia","volume":"68","author":"Hamada","year":"2013","journal-title":"Clinics"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1016\/S0888-7543(03)00129-0","article-title":"Identification of eight members of the Argonaute family in the human genome\u2606","volume":"82","author":"Sasaki","year":"2003","journal-title":"Genomics"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/S0925-4773(01)00499-3","article-title":"Two mouse piwi-related genes: Miwi and mili","volume":"108","author":"Kimura","year":"2001","journal-title":"Mech. Dev."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1016\/S1534-5807(02)00165-X","article-title":"miwi, a Murine Homolog of piwi, Encodes a Cytoplasmic Protein Essential for Spermatogenesis","volume":"2","author":"Deng","year":"2002","journal-title":"Dev. Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1242\/dev.00973","article-title":"Mili, a mammalian member ofpiwifamily gene, is essential for spermatogenesis","volume":"131","author":"Kimura","year":"2004","journal-title":"Development"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1016\/j.devcel.2007.03.001","article-title":"MIWI2 Is Essential for Spermatogenesis and Repression of Transposons in the Mouse Male Germline","volume":"12","author":"Carmell","year":"2007","journal-title":"Dev. Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"260","DOI":"10.4103\/aja.aja_58_17","article-title":"Altered PIWI-LIKE 1 and PIWI-LIKE 2 mRNA expression in ejaculated spermatozoa of men with impaired sperm characteristics","volume":"20","author":"Behre","year":"2018","journal-title":"Asian J. Androl."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"61","DOI":"10.5653\/cerm.2011.38.2.61","article-title":"Functions of PIWI proteins in spermatogenesis","volume":"38","author":"Bak","year":"2011","journal-title":"Clin. Exp. Reprod. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1530\/REP-16-0586","article-title":"Expression patterns of HENMT1 and PIWIL1 in human testis: Implications for transposon expression","volume":"154","author":"Hempfling","year":"2017","journal-title":"Reproduction"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1177\/1933719117725822","article-title":"Spermatozoa Expression of piR-31704, piR-39888, and piR-40349 and Their Correlation to Sperm Concentration and Fertilization Rate after ICSI","volume":"25","author":"Cui","year":"2017","journal-title":"Reprod. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2443","DOI":"10.1093\/humrep\/dex317","article-title":"Alterations in the sperm histone-retained epigenome are associated with unexplained male factor infertility and poor blastocyst development in donor oocyte IVF cycles","volume":"32","author":"Denomme","year":"2017","journal-title":"Hum. Reprod."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1090","DOI":"10.1016\/j.cell.2017.04.034","article-title":"Ubiquitination-Deficient Mutations in Human Piwi Cause Male Infertility by Impairing Histone-to-Protamine Exchange during Spermiogenesis","volume":"169","author":"Gou","year":"2017","journal-title":"Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1941","DOI":"10.1016\/j.cell.2021.03.001","article-title":"Lack of evidence for a role of PIWIL1 variants in human male infertility","volume":"184","author":"Oud","year":"2021","journal-title":"Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1002\/j.1939-4640.2005.tb02874.x","article-title":"A suite of novel human spermatozoal RNAs","volume":"26","author":"Ostermeier","year":"2005","journal-title":"J. Androl."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1093\/humupd\/dmt031","article-title":"The presence, role and clinical use of spermatozoal RNAs","volume":"19","author":"Jodar","year":"2013","journal-title":"Hum. Reprod. Update"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3819","DOI":"10.1242\/dev.099184","article-title":"The nuage mediates retrotransposon silencing in mouse primordial ovarian follicles","volume":"140","author":"Lim","year":"2013","journal-title":"Development"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Kohlrausch, F.B., Berteli, T.S., Wang, F., Navarro, P.A., and Keefe, D.L. (2021). Control of LINE-1 Expression Maintains Genome Integrity in Germline and Early Embryo Development. Reprod. Sci., 1\u201313.","DOI":"10.1007\/s43032-021-00461-1"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/21\/11320\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:19:02Z","timestamp":1760167142000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/21\/11320"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,20]]},"references-count":33,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["ijms222111320"],"URL":"https:\/\/doi.org\/10.3390\/ijms222111320","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,20]]}}}