{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T18:27:55Z","timestamp":1775240875422,"version":"3.50.1"},"reference-count":49,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2022,7,14]],"date-time":"2022-07-14T00:00:00Z","timestamp":1657756800000},"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>Recently it was shown that a specific form of male infertility in Holstein cattle was caused by a nonsense variant in the \u03b1\/\u03b2-hydrolase domain-containing 16B (ABHD16B) gene resulting in a protein truncation at amino acid position 218 (p.218Q*) and loss of function. Lipidomics showed that the absence of ABHD16B influenced the content of phosphatidylcholine (PC), ceramide (Cer), diacylglycerol (DAG), and sphingomyelin (SM) in variant carrier sperm membranes. However, the exact cause of infertility in affected sires has remained unclear until now. To elucidate the cause of infertility, we analyzed (i) standard sperm parameters (i.e., total sperm number, morphological intact sperm, total sperm motility), (ii) in vitro fertilizability and effects on early embryonic development, and (iii) sperm survival rates (i.e., capacitation time). The affected spermatozoa showed no changes in the usual sperm parameters and were also capable of fertilization in vitro. Furthermore, the absence of ABHD16B did not affect early embryonic development. Based on these results, it was concluded that the affected spermatozoa appeared to be fertilizable per se. Consequently, the actual cause of the inability to fertilize could only be due to a time- and\/or place-dependent process after artificial insemination and before fertilization. A process fundamental to the ability to fertilize after insemination is capacitation. Capacitation is a biochemical maturation process that spermatozoa undergo in the female genital tract and is inevitable for the successful fertilization of the oocyte. It is known that the presence and concentration of certain sperm membrane lipids are essential for the correct course of capacitation. However, precisely these lipids are absent in the membrane of spermatozoa affected by the ABHD16B truncation. Since all other causes of fertilization inability were excluded in the previous experiments, consequently, the only remaining hypothesis was that the loss of function of ABHD16B leads to a capacitation disruption. We were able to show that heterozygous and homozygous affected spermatozoa exhibit premature capacitation and therefore decay before fertilization. This effect of the loss of function of ABHD16B has not been described before and our studies now revealed why sires harboring the variant in the ABHD16B gene are infertile.<\/jats:p>","DOI":"10.3390\/ijms23147777","type":"journal-article","created":{"date-parts":[[2022,7,15]],"date-time":"2022-07-15T00:17:10Z","timestamp":1657844230000},"page":"7777","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["\u03b1\/\u03b2-Hydrolase D16B Truncation Results in Premature Sperm Capacitation in Cattle"],"prefix":"10.3390","volume":"23","author":[{"given":"Shuwen","family":"Shan","sequence":"first","affiliation":[{"name":"Institute of Veterinary Medicine, University of Goettingen, 37077 Goettingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fangzheng","family":"Xu","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Medicine, University of Goettingen, 37077 Goettingen, Germany"},{"name":"Malaria Functional Genomics Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Disease, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marc","family":"Hirschfeld","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Medicine, University of Goettingen, 37077 Goettingen, Germany"},{"name":"MVZ Brustzentrum Freiburg, 79110 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudia","family":"Herrmann","sequence":"additional","affiliation":[{"name":"Institute for Reproduction of Farm Animals Schoenow, 16321 Bernau, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7932-9412","authenticated-orcid":false,"given":"Martin","family":"Schulze","sequence":"additional","affiliation":[{"name":"Institute for Reproduction of Farm Animals Schoenow, 16321 Bernau, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ahmad Reza","family":"Sharifi","sequence":"additional","affiliation":[{"name":"Department of Animal Sciences, Devision Animal Breeding and Genetics, University of Goettingen, 37017 Goettingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Hoelker","sequence":"additional","affiliation":[{"name":"Department of Animal Sciences, Division Biotechnology and Reproduction of Farm Animals, University of Goettingen, 37077 Goettingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7635-9656","authenticated-orcid":false,"given":"Bertram","family":"Brenig","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Medicine, University of Goettingen, 37077 Goettingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1038\/168697b0","article-title":"Fertilizing capacity of spermatozoa deposited into the fallopian tubes","volume":"168","author":"Chang","year":"1951","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1071\/BI9510581","article-title":"Observations on the penetration of the sperm into the mammalian egg","volume":"4","author":"Austin","year":"1951","journal-title":"Aust. J. Biol. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"A125","DOI":"10.2741\/A125","article-title":"Human sperm activation during capacitation and acrosome reaction: Role of calcium, protein phosphorylation and lipid remodelling pathways","volume":"1","author":"Baldi","year":"1996","journal-title":"Front. Biosci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"7560","DOI":"10.1073\/pnas.78.12.7560","article-title":"Timing of fertilization in mammals: Sperm cholesterol\/phospholipid ratio as a determinant of the capacitation interval","volume":"78","author":"Davis","year":"1981","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1016\/j.theriogenology.2004.09.016","article-title":"Bicarbonate-induced membrane processing in sperm capacitation","volume":"63","author":"Harrison","year":"2005","journal-title":"Theriogenology"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"768","DOI":"10.1095\/biolreprod59.4.768","article-title":"Major proteins of bovine seminal plasma and high-density lipoprotein induce cholesterol efflux from epididymal sperm","volume":"59","author":"Moreau","year":"1998","journal-title":"Biol. Reprod."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Shan, S., Xu, F., Hirschfeld, M., and Brenig, B. (2021). Sperm Lipid Markers of Male Fertility in Mammals. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22168767"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1530\/jrf.0.0230193","article-title":"The movement of golden hamster spermatozoa before and after capacitation","volume":"23","author":"Yanagimachi","year":"1970","journal-title":"Reproduction"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1093\/humrep\/13.suppl_1.9","article-title":"Sperm capacitation and the acrosome reaction","volume":"13","author":"Fraser","year":"1998","journal-title":"Hum. Reprod."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1191","DOI":"10.1095\/biolreprod36.5.1191","article-title":"Initiation of hyperactivated flagellar bending in mouse sperm within the female reproductive tract","volume":"36","author":"Osman","year":"1987","journal-title":"Biol. Reprod."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1095\/biolreprod46.5.779","article-title":"Hyperactivated sperm progress in the mouse oviduct","volume":"46","author":"Demott","year":"1992","journal-title":"Biol. Reprod."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1095\/biolreprod44.2.375","article-title":"Evidence for the function of hyperactivated motility in sperm","volume":"44","author":"Suarez","year":"1991","journal-title":"Biol. Reprod."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"686","DOI":"10.1095\/biolreprod46.4.686","article-title":"Hyperactivation enhances mouse sperm capacity for penetrating viscoelastic media","volume":"46","author":"Suarez","year":"1992","journal-title":"Biol. Reprod."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/0012-1606(88)90437-X","article-title":"Hamster sperm penetration of the zona pellucida: Kinematic analysis and mechanical implications","volume":"130","author":"Drobnis","year":"1988","journal-title":"Dev. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1280","DOI":"10.1095\/biolreprod53.6.1280","article-title":"Sperm motility hyperactivation facilitates penetration of the hamster zona pellucida","volume":"53","author":"Stauss","year":"1995","journal-title":"Biol. Reprod."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Okabe, M. (2016). The acrosome reaction: A historical perspective. Sperm Acrosome Biogenesis and Function during Fertilization, Springer.","DOI":"10.1007\/978-3-319-30567-7_1"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1093\/molehr\/3.3.175","article-title":"Capacitation as a regulatory event that primes spermatozoa for the acrosome reaction and fertilization","volume":"3","author":"Leclerc","year":"1997","journal-title":"Mol. Hum. Reprod."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1093\/humupd\/dmn029","article-title":"Control of hyperactivation in sperm","volume":"14","author":"Suarez","year":"2008","journal-title":"Hum. Reprod. Update"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.1016\/S0015-0282(16)55992-5","article-title":"Human sperm hyperactivation in whole semen and its association with low superoxide scavenging capacity in seminal plasma","volume":"59","author":"Gagnon","year":"1993","journal-title":"Fertil. Steril."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1002\/j.1939-4640.2000.tb03268.x","article-title":"Semen cryopreservation in domestic animals: A damaging and capacitating phenomenon","volume":"21","author":"Bailey","year":"2000","journal-title":"J. Androl."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1002\/j.1939-4640.1997.tb01953.x","article-title":"Premature capacitation of bovine spermatozoa is initiated by cryopreservation","volume":"18","author":"Cormier","year":"1997","journal-title":"J. Androl."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1046\/j.1439-0272.2003.00564.x","article-title":"Reactive oxygen species induce reversible capacitation in human spermatozoa","volume":"35","author":"Villegas","year":"2003","journal-title":"Andrologia"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"110512","DOI":"10.1016\/j.ecoenv.2020.110512","article-title":"Bisphenol A affects the maturation and fertilization competence of spermatozoa","volume":"196","author":"Park","year":"2020","journal-title":"Ecotoxicol. Environ. Saf."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1002\/j.1939-4640.2001.tb02228.x","article-title":"Semenogelin, the main protein of semen coagulum, inhibits human sperm capacitation by interfering with the superoxide anion generated during this process","volume":"22","author":"Yoshida","year":"2001","journal-title":"J. Androl."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1038\/aja.2009.6","article-title":"NYD-SP27, a novel intrinsic decapacitation factor in sperm","volume":"11","author":"Bi","year":"2009","journal-title":"Asian J. Androl."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Shan, S., Xu, F., Bleyer, M., Becker, S., Melbaum, T., Wemheuer, W., Hirschfeld, M., Wacker, C., Zhao, S., and Sch\u00fctz, E. (2020). Association of \u03b1\/\u03b2-Hydrolase D16B with Bovine Conception Rate and Sperm Plasma Membrane Lipid Composition. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21020627"},{"key":"ref_27","first-page":"792","article-title":"Mammalian alpha beta hydrolase domain (ABHD) proteins: Lipid metabolizing enzymes at the interface of cell signaling and energy metabolism","volume":"1831","author":"Lord","year":"2013","journal-title":"Biochim. Biophys. Acta (BBA)-Mol. Cell Biol. Lipids"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1126\/science.aad6887","article-title":"Unconventional endocannabinoid signaling governs sperm activation via the sex hormone progesterone","volume":"352","author":"Miller","year":"2016","journal-title":"Science"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1886","DOI":"10.1016\/j.biochi.2010.08.015","article-title":"Differential involvement of rat sperm choline glycerophospholipids and sphingomyelin in capacitation and the acrosomal reaction","volume":"92","author":"Zanetti","year":"2010","journal-title":"Biochimie"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"118704","DOI":"10.1016\/j.bbamcr.2020.118704","article-title":"Ceramide induces a multicomponent intracellular calcium increase triggering the acrosome secretion in human sperm","volume":"1867","author":"Vaquer","year":"2020","journal-title":"Biochim. Biophys. Acta (BBA)-Mol. Cell Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1002\/(SICI)1098-2795(199709)48:1<95::AID-MRD12>3.0.CO;2-V","article-title":"Diacylglycerol species as messengers and substrates for phosphatidylcholine re-synthesis during Ca2+-dependent exocytosis in boar spermatozoa","volume":"48","author":"Vazquez","year":"1997","journal-title":"Mol. Reprod. Dev. Inc. Gamete Res."},{"key":"ref_32","first-page":"1186","article-title":"Diacylglycerol stimulates acrosomal exocytosis by feeding into a PKC-and PLD1-dependent positive loop that continuously supplies phosphatidylinositol 4, 5-bisphosphate","volume":"1821","author":"Lopez","year":"2012","journal-title":"Biochim. Biophys. Acta (BBA)-Mol. Cell Biol. Lipids"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1530\/jrf.0.0990135","article-title":"Ca2+-related changes in the capacitation state of human spermatozoa assessed by a chlortetracycline fluorescence assay","volume":"99","author":"DasGupta","year":"1993","journal-title":"Reproduction"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"191","DOI":"10.4103\/0974-1208.170370","article-title":"Trends of male factor infertility, an important cause of infertility: A review of literature","volume":"8","author":"Kumar","year":"2015","journal-title":"J. Hum. Reprod. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1111\/j.1365-2605.2005.00630.x","article-title":"Sperm function tests and fertility","volume":"29","author":"Aitken","year":"2006","journal-title":"Int. J. Androl."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1046\/j.1439-0531.2003.00436.x","article-title":"Laboratory semen assessment and prediction of fertility: Still utopia?","volume":"38","year":"2003","journal-title":"Reprod. Domest. Anim."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/j.rbmo.2017.12.005","article-title":"A search for molecular mechanisms underlying male idiopathic infertility","volume":"36","author":"Bracke","year":"2018","journal-title":"Reprod. Biomed. Online"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2671","DOI":"10.1091\/mbc.e11-12-1025","article-title":"Mice expressing aberrant sperm-specific protein PMIS2 produce normal-looking but fertilization-incompetent spermatozoa","volume":"23","author":"Yamaguchi","year":"2012","journal-title":"Mol. Biol. Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4198","DOI":"10.1096\/fj.12-205211","article-title":"Epididymal protein Rnase10 is required for post-testicular sperm maturation and male fertility","volume":"26","author":"Krutskikh","year":"2012","journal-title":"FASEB J."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1095\/biolreprod28.5.1043","article-title":"Phospholipid methylation increases during capacitation of golden hamster sperm in vitro","volume":"28","author":"Llanos","year":"1983","journal-title":"Biol. Reprod."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"269","DOI":"10.2527\/jas1975.402269x","article-title":"Alteration of phospholipids in porcine spermatozoa during in vivo uterus and oviduct incubation","volume":"40","author":"Snider","year":"1975","journal-title":"J. Anim. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1095\/biolreprod63.4.1129","article-title":"Sphingomyelin modulates capacitation of human sperm in vitro","volume":"63","author":"Cross","year":"2000","journal-title":"Biol. Reprod."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1177\/1933719113504467","article-title":"The pattern of tyrosine phosphorylation in human sperm in response to binding to zona pellucida or hyaluronic acid","volume":"21","author":"Sati","year":"2014","journal-title":"Reprod. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1242\/dev.121.4.1129","article-title":"Capacitation of mouse spermatozoa. I. Correlation between the capacitation state and protein tyrosine phosphorylation","volume":"121","author":"Visconti","year":"1995","journal-title":"Development"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1016\/j.anireprosci.2014.06.008","article-title":"Influences on semen traits used for selection of young AI boars","volume":"148","author":"Schulze","year":"2014","journal-title":"Anim. Reprod. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.theriogenology.2021.09.024","article-title":"Suitability of semen stress tests for predicting fertilizing capacity of boar ejaculates","volume":"176","author":"Schulze","year":"2021","journal-title":"Theriogenology"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1171","DOI":"10.1095\/biolreprod38.5.1171","article-title":"Capacitation of bovine sperm by heparin","volume":"38","author":"Parrish","year":"1988","journal-title":"Biol. Reprod."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1016\/S0093-691X(99)00162-4","article-title":"High bovine blastocyst development in a static in vitro production system using SOFaa medium supplemented with sodium citrate and myo-inositol with or without serum-proteins","volume":"52","author":"Holm","year":"1999","journal-title":"Theriogenology"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1002\/mrd.1080400213","article-title":"Ca2+-regulating mechanisms that modulate bull sperm capacitation and acrosomal exocytosis as determined by chlortetracycline analysis","volume":"40","author":"Fraser","year":"1995","journal-title":"Mol. Reprod. Dev."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/14\/7777\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:50:24Z","timestamp":1760140224000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/14\/7777"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,14]]},"references-count":49,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["ijms23147777"],"URL":"https:\/\/doi.org\/10.3390\/ijms23147777","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,14]]}}}