{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T19:35:55Z","timestamp":1790105755195,"version":"4.0.1"},"reference-count":64,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,7,6]],"date-time":"2021-07-06T00:00:00Z","timestamp":1625529600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell Dev. Biol."],"abstract":"<jats:p>Cation channel of sperm (CatSper), the main sperm-specific Ca<jats:sup>2+<\/jats:sup> channel, plays a key role in mammalian fertilization, and it is essential for male fertility, becoming an attractive target for contraception. Based on this, in the present work, we investigated the effects of CatSper inactivation on <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic> sperm fertilizing ability and the mechanisms underlying such effects. Exposure of cauda epididymal mouse sperm to different concentrations (1\u201320 \u03bcM) of the potent CatSper inhibitor HC-056456 (HC) during <jats:italic>in vitro<\/jats:italic> capacitation showed no effects on sperm viability but significantly affected Ca<jats:sup>2+<\/jats:sup> entry into the cells, progressive motility, protein tyrosine phosphorylation, induced acrosome reaction, and hyperactivation, as well as the sperm\u2019s ability to <jats:italic>in vitro<\/jats:italic> fertilize cumulus oocyte complexes and zona-free eggs. Whereas the presence of HC during gamete coincubation did not affect <jats:italic>in vitro<\/jats:italic> fertilization, exposure of either non-capacitating or already capacitated sperm to HC prior to gamete coincubation severely reduced fertilization, indicating that sperm function is affected by HC when the cells are incubated with the drug before sperm\u2013egg interaction. Of note, insemination of HC-treated sperm into the uterus significantly or completely reduced the percentage of oviductal fertilized eggs showing, for the first time, the effects of a CatSper inhibitor on <jats:italic>in vivo<\/jats:italic> fertilization. These observations, together with the finding that HC affects sperm fertilizing ability independently of the sperm capacitation status, provide further insights on how CatSper regulates sperm function and represent a solid proof of concept for developing a male\/female non-hormonal contraceptive based on the pharmacological blockage of CatSper activity.<\/jats:p>","DOI":"10.3389\/fcell.2021.686461","type":"journal-article","created":{"date-parts":[[2021,7,6]],"date-time":"2021-07-06T05:45:18Z","timestamp":1625550318000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":26,"title":["Pharmacological Inactivation of CatSper Blocks Sperm Fertilizing Ability Independently of the Capacitation Status of the Cells: Implications for Non-hormonal Contraception"],"prefix":"10.3389","volume":"9","author":[{"given":"Ludmila","family":"Curci","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guillermo","family":"Carvajal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valeria","family":"Sulzyk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soledad Natalia","family":"Gonzalez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia S.","family":"Cuasnic\u00fa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,7,6]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"228","DOI":"10.1111\/cts.12708","article-title":"Development of novel male contraceptives.","volume":"13","author":"Amory","year":"2020","journal-title":"Clin. Transl. Sci."},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1038\/170326a0","article-title":"The capacitation of the mammalian sperm.","volume":"170","author":"Austin","year":"1952","journal-title":"Nature"},{"key":"B3","doi-asserted-by":"publisher","first-page":"505","DOI":"10.1016\/j.ajhg.2009.03.004","article-title":"Human male infertility caused by mutations in the CATSPER1 channel protein.","volume":"84","author":"Avenarius","year":"2009","journal-title":"Am. J. Hum. Genet."},{"key":"B4","doi-asserted-by":"publisher","first-page":"497","DOI":"10.1038\/sj.ejhg.5200991","article-title":"CATSPER2, a human autosomal nonsyndromic male infertility gene.","volume":"11","author":"Avidan","year":"2003","journal-title":"Eur. J. Hum. Genet."},{"key":"B5","doi-asserted-by":"publisher","first-page":"1825","DOI":"10.1007\/s10695-020-00829-1","article-title":"The voltage-gated T-type Ca2+ channel is key to the sperm motility of Atlantic salmon (Salmo salar).","volume":"46","author":"Beltr\u00e1n","year":"2020","journal-title":"Fish Physiol. Biochem."},{"key":"B6","doi-asserted-by":"publisher","first-page":"807","DOI":"10.1095\/biolreprod.105.042184","article-title":"Mice deficient in CHRNA7, a subunit of the nicotinic acetylcholine receptor, produce sperm with impaired motility.","volume":"73","author":"Bray","year":"2005","journal-title":"Biol. Reproduct."},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.01438","article-title":"The Ca2+-activated K+ current of human sperm is mediated by Slo3.","volume":"3","author":"Brenker","year":"2014","journal-title":"ELife"},{"key":"B8","doi-asserted-by":"publisher","first-page":"1812","DOI":"10.1093\/humrep\/dey278","article-title":"Homozygous in-frame deletion in CATSPERE in aman producing spermatozoa with loss of CatSper function and compromised fertilizing capacity.","volume":"33","author":"Brown","year":"2018","journal-title":"Hum. Reproduct."},{"key":"B9","doi-asserted-by":"publisher","first-page":"240","DOI":"10.1093\/molehr\/gaw005","article-title":"Fertilization defects in sperm from Cysteine-rich secretory protein 2 (Crisp2) knockout mice: implications for fertility disorders.","volume":"22","author":"Brukman","year":"2016","journal-title":"Mol. Hum. Reproduct."},{"key":"B10","doi-asserted-by":"publisher","first-page":"5276","DOI":"10.1002\/jcp.27337","article-title":"Tyrosine phosphorylation signaling regulates Ca2+ entry by affecting intracellular pH during human sperm capacitation.","volume":"234","author":"Brukman","year":"2019","journal-title":"J. Cell. Physiol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"2735","DOI":"10.1093\/molbev\/msu218","article-title":"Early evolution of the eukaryotic Ca2 + signaling machinery: conservation of the CatSper channel complex.","volume":"31","author":"Cai","year":"2014","journal-title":"Mol. Biol. Evol."},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0006844","article-title":"Pharmacological targeting of native CatSper channels reveals a required role in maintenance of sperm hyperactivation.","volume":"4","author":"Carlson","year":"2009","journal-title":"PLoS One"},{"key":"B13","doi-asserted-by":"publisher","first-page":"14864","DOI":"10.1073\/pnas.2536658100","article-title":"CatSper1 required for evoked Ca2+ entry and control of flagellar function in sperm.","volume":"100","author":"Carlson","year":"2003","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-35719-3","article-title":"Impaired male fertility and abnormal epididymal epithelium differentiation in mice lacking CRISP1 and CRISP4.","volume":"8","author":"Carvajal","year":"2018","journal-title":"Sci. Rep."},{"key":"B15","doi-asserted-by":"publisher","first-page":"697","DOI":"10.1016\/j.biortech.2006.10.008","article-title":"Fertilizing capacity of spermatozoa deposited into the fallopian tubes.","volume":"168","author":"Chang","year":"1951","journal-title":"Nature"},{"key":"B16","doi-asserted-by":"publisher","first-page":"4735","DOI":"10.1002\/jcp.26262","article-title":"Acrosomal alkalization triggers Ca 2+ release and acrosome reaction in mammalian spermatozoa.","volume":"233","author":"Ch\u00e1vez","year":"2018","journal-title":"J. Cell. Physiol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1038\/ncomms1153","article-title":"A novel gene required for male fertility and functional CATSPER channel formation in spermatozoa.","volume":"2","author":"Chung","year":"2011","journal-title":"Nat. Commun."},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.23082","article-title":"Catsper\u03c2 regulates the structural continuity of sperm ca2+vsignaling domains and is required for normal fertility.","volume":"6","author":"Chung","year":"2017","journal-title":"ELife"},{"key":"B19","doi-asserted-by":"publisher","first-page":"808","DOI":"10.1016\/j.cell.2014.02.056","article-title":"Structurally distinct Ca(2+) signaling domains of sperm flagella orchestrate tyrosine phosphorylation and motility.","volume":"157","author":"Chung","year":"2014","journal-title":"Cell"},{"key":"B20","doi-asserted-by":"publisher","first-page":"1325","DOI":"10.1095\/biolreprod.103.022822","article-title":"Bicarbonate is required for migration of sperm epididymal protein DE (CRISP-1) to the equatorial segment and expression of rat sperm fusion ability.","volume":"70","author":"Da Ros","year":"2004","journal-title":"Biol. Reproduct."},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.62043","article-title":"3D in situ imaging of female reproductive tract reveals molecular signatures of fertilizing spermatozoa in mice.","volume":"9","author":"Ded","year":"2020","journal-title":"ELife"},{"key":"B22","doi-asserted-by":"publisher","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. Reproduct."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1186\/s12610-018-0078-y","article-title":"Epididymal approaches to male contraception.","volume":"28","author":"Drevet","year":"2018","journal-title":"Basic Clin. Androl."},{"key":"B24","doi-asserted-by":"publisher","first-page":"1029","DOI":"10.1095\/biolreprod59.5.1029","article-title":"Potential contraceptive use of epididymal proteins: immunization of male rats with epididymal protein DE inhibits sperm fusion ability.","volume":"59","author":"Ellerman","year":"1998","journal-title":"Biol. Reproduct."},{"key":"B25","doi-asserted-by":"publisher","first-page":"1213","DOI":"10.1083\/jcb.201412041","article-title":"CRISP1 as a novel CatSper regulator that modulates sperm motility and orientation during fertilization.","volume":"210","author":"Ernesto","year":"2015","journal-title":"J. Cell Biol."},{"key":"B26","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1016\/0378-4320(78)90008-8","article-title":"The relationship between sperm concentration and fertilization in vitro of mouse eggs.","volume":"13","author":"Fraser","year":"1975","journal-title":"Biol. Reproduct."},{"key":"B27","doi-asserted-by":"publisher","first-page":"345","DOI":"10.1071\/RD08183","article-title":"CatSper-null mutant spermatozoa are unable to ascend beyond the oviductal reservoir.","volume":"21","author":"Ho","year":"2009","journal-title":"Reproduct. Fertility Dev."},{"key":"B28","doi-asserted-by":"crossref","first-page":"1480","DOI":"10.1016\/j.cell.2019.03.047","article-title":"Dual sensing of physiologic pH and calcium by EFCAB9 regulates sperm motility.","volume":"177","author":"Hwang","year":"2019","journal-title":"Cell"},{"key":"B29","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1095\/biolreprod.107.060186","article-title":"Catsper3 and Catsper4 are essential for sperm hyperactivated motility and male fertility in the mouse.","volume":"77","author":"Jin","year":"2007","journal-title":"Biol. Reproduct."},{"key":"B30","doi-asserted-by":"publisher","first-page":"9.8.1","DOI":"10.1002\/0471142956.cy0908s72","article-title":"Measurement of intracellular ions by flow cytometry.","volume":"72","author":"June","year":"1997","journal-title":"Curr. Protoc. Cytom."},{"key":"B31","doi-asserted-by":"publisher","first-page":"737","DOI":"10.1038\/nature04417","article-title":"Whole-cell patch-clamp measurements of spermatozoa reveal an alkaline-activated Ca2+ channel.","volume":"439","author":"Kirichok","year":"2006","journal-title":"Nature"},{"key":"B32","doi-asserted-by":"publisher","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of structural proteins during the assembly of the head of bacteriophage T4.","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nat. Publishing Group"},{"key":"B33","doi-asserted-by":"publisher","first-page":"816","DOI":"10.1016\/j.tibs.2016.08.002","article-title":"Contraception: search for an ideal unisex mechanism by targeting ion channels.","volume":"41","author":"Lishko","year":"2016","journal-title":"Trends Biochem. Sci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"387","DOI":"10.1038\/nature09767","article-title":"Progesterone activates the principal Ca2+ channel of human sperm.","volume":"471","author":"Lishko","year":"2011","journal-title":"Nature"},{"key":"B35","doi-asserted-by":"publisher","first-page":"4667","DOI":"10.1113\/jphysiol.2010.194142","article-title":"The role of Hv1 and CatSper channels in sperm activation.","volume":"588","author":"Lishko","year":"2010","journal-title":"J. Physiol."},{"key":"B36","doi-asserted-by":"publisher","first-page":"18945","DOI":"10.1074\/jbc.M701083200","article-title":"CatSper\u03b2, a novel transmembrane protein in the CatSper channel complex.","volume":"282","author":"Liu","year":"2007","journal-title":"J. Biol. Chem."},{"key":"B37","doi-asserted-by":"publisher","first-page":"414","DOI":"10.1093\/humrep\/dey377","article-title":"A novel copy number variation in CATSPER2 causes idiopathic male infertility with normal semen parameters.","volume":"34","author":"Luo","year":"2019","journal-title":"Hum. Reproduct."},{"key":"B38","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1016\/j.reprotox.2016.02.003","article-title":"Matrine compromises mouse sperm functions by a [Ca2+]i-related mechanism.","volume":"60","author":"Luo","year":"2016","journal-title":"Reproduct. Toxicol."},{"key":"B39","doi-asserted-by":"publisher","first-page":"5743","DOI":"10.1073\/pnas.1700367114","article-title":"Regulation of the sperm calcium channel CatSper by endogenous steroids and plant triterpenoids.","volume":"114","author":"Mannowetz","year":"2017","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B40","doi-asserted-by":"publisher","first-page":"1758","DOI":"10.1002\/jcp.24873","article-title":"Biphasic role of calcium in mouse sperm capacitation signaling pathways.","volume":"230","author":"Navarrete","year":"2015","journal-title":"J. Cell. Physiol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"7688","DOI":"10.1073\/pnas.0702018104","article-title":"KSper, a pH-sensitive K current that controls sperm membrane potential.","volume":"104","author":"Navarro","year":"2007","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B42","doi-asserted-by":"publisher","first-page":"263","DOI":"10.1083\/jcb.66.2.263","article-title":"Ultrastructural localization of lectin-binding sites on the zonae pellucidae and plasma membranes of mammalian eggs.","volume":"66","author":"Nicolson","year":"1975","journal-title":"J. Cell Biol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"1219","DOI":"10.1073\/pnas.0610286104","article-title":"All four CatSper ion channel proteins are required for male fertility and sperm cell hyperactivated motility.","volume":"104","author":"Qi","year":"2007","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B44","doi-asserted-by":"publisher","first-page":"14869","DOI":"10.1073\/pnas.2136654100","article-title":"Hyperactivated sperm motility driven by CatSper2 is required for fertilization.","volume":"100","author":"Quill","year":"2003","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B45","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1038\/35098027","article-title":"A sperm ion channel required for sperm motility and male fertility.","volume":"413","author":"Ren","year":"2001","journal-title":"Nature"},{"key":"B46","doi-asserted-by":"publisher","first-page":"3144","DOI":"10.1111\/bph.14355","article-title":"A novel cross-species inhibitor to study the function of CatSper Ca2+ channels in sperm.","volume":"175","author":"Rennhack","year":"2018","journal-title":"Br. J. Pharmacol."},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-397175-3.00017-X","article-title":"The epididymis","author":"Robaire","year":"2014","journal-title":"Knobil and Neill\u2019s Physiology of Reproduction: Two-Volume Set"},{"key":"B48","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1002\/mrd.1080260110","article-title":"Ca2+ uptake during capacitation of mouse spermatozoa and the effect of an anion transport inhibitor on Ca2+ uptake.","volume":"26","author":"Ruknudin","year":"1990","journal-title":"Mol. Reproduct. Dev."},{"key":"B49","doi-asserted-by":"publisher","first-page":"1041","DOI":"10.1016\/j.febslet.2010.02.005","article-title":"The SLO3 sperm-specific potassium channel plays a vital role in male fertility.","volume":"584","author":"Santi","year":"2010","journal-title":"FEBS Lett."},{"key":"B50","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1016\/j.rbmo.2014.09.014","article-title":"CatSper channel, sperm function and male fertility.","volume":"30","author":"Singh","year":"2015","journal-title":"Reproduct. BioMed. Online"},{"key":"B51","doi-asserted-by":"publisher","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. Reproduct."},{"key":"B52","doi-asserted-by":"publisher","first-page":"9435","DOI":"10.1074\/jbc.RA118.002286","article-title":"Disruption of protein kinase A localization induces acrosomal exocytosis in capacitated mouse sperm.","volume":"293","author":"Stival","year":"2018","journal-title":"J. Biol. Chem."},{"key":"B53","doi-asserted-by":"publisher","first-page":"382","DOI":"10.1038\/nature09769","article-title":"The CatSper channel mediates progesterone-induced Ca2+ influx in human sperm.","volume":"471","author":"Str\u00fcnker","year":"2011","journal-title":"Nature"},{"key":"B54","doi-asserted-by":"publisher","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. Reproduct."},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.115.129783","article-title":"Progesterone accelerates the completion of sperm capacitation and activates catSper channel in spermatozoa from the Rhesus macaque.","volume":"93","author":"Sumigama","year":"2015","journal-title":"Biol. Reproduct."},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1186\/s12958-017-0281-2","article-title":"The Catsper channel and its roles in male fertility?: a systematic review.","volume":"15","author":"Sun","year":"2017","journal-title":"Reproduct. Biol. Endocrinol."},{"key":"B57","doi-asserted-by":"publisher","first-page":"418","DOI":"10.1093\/humrep\/det454","article-title":"The CatSper calcium channel in human sperm?: relation with motility and involvement in progesterone-induced acrosome reaction.","volume":"29","author":"Tamburrino","year":"2014","journal-title":"Hum. Reprod."},{"key":"B58","doi-asserted-by":"publisher","first-page":"3235","DOI":"10.1074\/jbc.274.5.3235","article-title":"Cholesterol efflux-mediated signal transduction in mammalian sperm: \u03b2- Cyclodextrins initiate transmembrane signaling leading to an increase in protein tyrosine phosphorylation and capacitation.","volume":"274","author":"Visconti","year":"1999","journal-title":"J. Biol. Chem."},{"key":"B59","doi-asserted-by":"publisher","first-page":"539","DOI":"10.1095\/biolreprod.109.077107","article-title":"A novel, single, transmembrane protein CATSPERG is associated with CATSPER1 channel protein.","volume":"81","author":"Wang","year":"2009","journal-title":"Biol. Reprod."},{"key":"B60","doi-asserted-by":"publisher","first-page":"46","DOI":"10.1038\/s41585-020-00390-9","article-title":"Sperm ion channels and transporters in male fertility and infertility.","volume":"18","author":"Wang","year":"2021","journal-title":"Nat. Rev. Urol."},{"key":"B61","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/dev243","article-title":"Specific loss of CatSper function is sufficient to compromise fertilizing capacity of human spermatozoa.","volume":"30","author":"Williams","year":"2015","journal-title":"Hum. Reproduct."},{"key":"B62","doi-asserted-by":"publisher","DOI":"10.1186\/1477-7827-7-119","article-title":"The BSA-induced Ca(2+) influx during sperm capacitation is CATSPER channel-dependent.","volume":"7","author":"Xia","year":"2009","journal-title":"Reproduct. Biol. Endocrinol."},{"key":"B63","doi-asserted-by":"publisher","first-page":"5879","DOI":"10.1073\/pnas.1100240108","article-title":"Deletion of the Slo3 gene abolishes alkalization-activated K+ current in mouse spermatozoa.","volume":"108","author":"Zeng","year":"2011","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B64","doi-asserted-by":"publisher","first-page":"233","DOI":"10.1136\/jmg.2006.045765","article-title":"Sensorineural deafness and male infertility: a contiguous gene deletion syndrome.","volume":"44","author":"Zhang","year":"2007","journal-title":"J. Med. Genet."}],"container-title":["Frontiers in Cell and Developmental Biology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2021.686461\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,7,6]],"date-time":"2021-07-06T05:45:25Z","timestamp":1625550325000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2021.686461\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,6]]},"references-count":64,"alternative-id":["10.3389\/fcell.2021.686461"],"URL":"https:\/\/doi.org\/10.3389\/fcell.2021.686461","relation":{},"ISSN":["2296-634X"],"issn-type":[{"value":"2296-634X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,6]]},"article-number":"686461"}}