{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,27]],"date-time":"2026-06-27T13:19:53Z","timestamp":1782566393878,"version":"3.54.5"},"reference-count":41,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:00:00Z","timestamp":1764288000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Center for Scientific and Technological Research","award":["PIP 2903"],"award-info":[{"award-number":["PIP 2903"]}]},{"name":"Argentinian Ministry of Education","award":["V309"],"award-info":[{"award-number":["V309"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Cryopreservation of sperm is a resource used for artificial insemination. In the case of bovines, it ensures the reproduction of animals through cells from males with a highlighted genetic background. The fertilisation capacity of sperm cells is achieved after capacitation, a process that includes subcellular modifications that could be altered by cryopreservation. Intracellular calcium handling plays a crucial role in the development of capacitation, culminating in the acrosomal reaction. SERCA protein is responsible for calcium reuptake into the acrosome, which is one of the main calcium reservoirs of sperm cells. In this work, we studied the relationships between SERCA activity and sperm motility, capacitation progression, actin polymerisation, and intracellular calcium handling in cryopreserved bovine spermatozoa. Inhibition of SERCA activity reduced sperm motility and induced hyperactivation patterns. It also increased the proportion of cells with acrosomal reaction and earlier actin depolymerisation, an event necessary to induce the acrosomal reaction. All changes occurred in concordance with a significant increase in intracellular calcium concentration (Ca2+). Our findings suggest that a thapsigargin-sensitive Ca2+ pump consistent with SERCA activity remains responsive in cryopreserved spermatozoa from the bull studied, under specific cryopreservation and incubation conditions tested, and may contribute to Ca2+ handling, motility changes, and premature acrosomal exocytosis.<\/jats:p>","DOI":"10.3390\/cells14231892","type":"journal-article","created":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T11:19:51Z","timestamp":1764328791000},"page":"1892","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Role of the Ca2+-ATPase Pump (SERCA) in Capacitation and the Acrosome Reaction of Cryopreserved Bull Spermatozoa"],"prefix":"10.3390","volume":"14","author":[{"given":"Maia A.","family":"Rodr\u00edguez","sequence":"first","affiliation":[{"name":"Reprosemyx S.A., Calle 4 # 693, La Plata 1900, Argentina"},{"name":"Laboratorio de An\u00e1lisis de Im\u00e1genes, Universidad Nacional de La Plata, La Plata 1900, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro","family":"Orlowski","sequence":"additional","affiliation":[{"name":"Centro de Investigaciones Cardiovasculares \u201cDr. Horacio E. Cingolani\u201d, Facultad de Ciencias M\u00e9dicas, Universidad Nacional de La Plata\u2014Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas, Av 60 & 120 2do piso, La Plata 1900, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enrique L.","family":"Portiansky","sequence":"additional","affiliation":[{"name":"Laboratorio de An\u00e1lisis de Im\u00e1genes, Universidad Nacional de La Plata, La Plata 1900, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5582-642X","authenticated-orcid":false,"given":"Paola","family":"Ferrero","sequence":"additional","affiliation":[{"name":"Reprosemyx S.A., Calle 4 # 693, La Plata 1900, Argentina"},{"name":"Centro de Investigaciones Cardiovasculares \u201cDr. Horacio E. Cingolani\u201d, Facultad de Ciencias M\u00e9dicas, Universidad Nacional de La Plata\u2014Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas, Av 60 & 120 2do piso, La Plata 1900, Argentina"},{"name":"Departamento de Ciencias B\u00e1sicas y Experimentales, Escuela de Ciencias Agrarias, Naturales y Ambientales, Universidad Nacional del Noroeste de la Provincia de Buenos Aires (UNNOBA), Pergamino 2772, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1007\/978-3-319-30567-7_5","article-title":"Sperm Capacitation and Acrosome Reaction in Mammalian Sperm","volume":"220","author":"Stival","year":"2016","journal-title":"Adv. Anat. Embryol. Cell Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/B978-0-12-397175-3.00004-1","article-title":"Fertilization in Mammals","volume":"Volume 1","author":"Florman","year":"2015","journal-title":"Knobil and Neill\u2019s Physiology of Reproduction: Two-Volume Set"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Takei, G.L. (2024). Molecular mechanisms of mammalian sperm capacitation, and its regulation by sodium-dependent secondary active transporters. Reprod. Med. Biol., 23, Available online: https:\/\/pubmed.ncbi.nlm.nih.gov\/39416520\/.","DOI":"10.1002\/rmb2.12614"},{"key":"ref_4","first-page":"321","article-title":"Signaling pathways in sperm capacitation and acrosome reaction","volume":"49","author":"Breibart","year":"2003","journal-title":"Cell. Mol. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1002\/mrd.21393","article-title":"Heads or tails? Structural events and molecular mechanisms that promote mammalian sperm acrosomal exocytosis and motility","volume":"79","author":"Buffone","year":"2012","journal-title":"Mol. Reprod. Dev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1093\/molehr\/gau110","article-title":"Kinetics of human sperm acrosomal exocytosis","volume":"21","author":"Sosa","year":"2015","journal-title":"Mol. Hum. Reprod."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"28220","DOI":"10.1074\/jbc.M312599200","article-title":"Plasma membrane Ca2+ ATPase 4 is required for sperm motility and male fertility","volume":"279","author":"Schuh","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1111\/j.1365-2605.2009.01022.x","article-title":"Arrangement of PMCA4 in bovine sperm membrane fractions","volume":"33","author":"Post","year":"2010","journal-title":"Int. J. Androl."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1093\/biolre\/iox082","article-title":"Genomic identification, expression profiling, and functional characterization of CatSper channels in the bovine","volume":"97","author":"Johnson","year":"2017","journal-title":"Biol. Reprod."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1093\/molehr\/gam012","article-title":"Identification and localisation of SERCA 2 isoforms in mammalian sperm","volume":"13","author":"Lawson","year":"2007","journal-title":"Mol. Hum. Reprod."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"R65","DOI":"10.1530\/REP-15-0102","article-title":"Regulation and roles of Ca2+ stores in human sperm","volume":"150","author":"Correia","year":"2015","journal-title":"Reproduction"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Garriga, F., Mart\u00ednez-Hern\u00e1ndez, J., Parra-Balaguer, A., Llavanera, M., and Yeste, M. (2025). The Sarcoplasmic\/Endoplasmic reticulum Ca2+-ATPase (SERCA) is present in pig sperm and modulates their physiology over liquid preservation. Sci. Rep., 15, Available online: https:\/\/pubmed.ncbi.nlm.nih.gov\/39905176\/.","DOI":"10.1038\/s41598-025-88012-5"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Kato, Y., Kumar, S., Lessard, C., and Bailey, J.L. (2021). ACRBP (Sp32) is involved in priming sperm for the acrosome reaction and the binding of sperm to the zona pellucida in a porcine model. PLoS ONE, 16, Available online: https:\/\/pubmed.ncbi.nlm.nih.gov\/34086710\/.","DOI":"10.1371\/journal.pone.0251973"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"jcs215897","DOI":"10.1242\/jcs.215897","article-title":"The actin cytoskeleton of the mouse sperm flagellum is organized in a helical structure","volume":"131","author":"Gervasi","year":"2018","journal-title":"J. Cell. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1530\/rep.1.00269","article-title":"Role of actin cytoskeleton in mammalian sperm capacitation and the acrosome reaction","volume":"129","author":"Breitbart","year":"2005","journal-title":"Reproduction"},{"key":"ref_16","first-page":"129","article-title":"Role of Actin Cytoskeleton During Mammalian Sperm Acrosomal Exocytosis","volume":"Volume 220","author":"Romarowski","year":"2016","journal-title":"Sperm Acrosome Biogenesis and Function During Fertilization. Advances in Anatomy, Embryology and Cell Biology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1016\/j.bbrc.2017.11.001","article-title":"Actin cytoskeleton and sperm function","volume":"506","author":"Breitbart","year":"2018","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1095\/biolreprod.102.009233","article-title":"Remodeling of the actin cytoskeleton during mammalian sperm capacitation and acrosome reaction","volume":"68","author":"Brener","year":"2003","journal-title":"Biol. Reprod."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Benko, F., Mohammadi-Sangcheshmeh, A., \u010eura\u010dka, M., Luk\u00e1\u010d, N., and Tvrd\u00e1, E. (2022). In vitro versus cryo-induced capacitation of bovine spermatozoa, part 1: Structural, functional, and oxidative similarities and differences. PLoS ONE, 17, Available online: https:\/\/pubmed.ncbi.nlm.nih.gov\/36269791\/.","DOI":"10.1371\/journal.pone.0276683"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1071\/RD07170","article-title":"Cryopreservation affects bovine sperm intracellular parameters associated with capacitation and acrosome exocytosis","volume":"21","author":"Bailey","year":"2009","journal-title":"Reprod. Fertil. Dev."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1606","DOI":"10.1095\/biolreprod65.5.1606","article-title":"An inositol 1,4,5-trisphosphate receptor-gated intracellular Ca(2+) store is involved in regulating sperm hyperactivated motility","volume":"65","author":"Ho","year":"2001","journal-title":"Biol. Reprod."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-020-72425-5","article-title":"Cyclic AMP efflux through MRP4 regulates actin dynamics signalling pathway and sperm motility in bovines","volume":"10","author":"Chiarante","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1111\/j.1365-2605.2003.00438.x","article-title":"Effects of Ca-ATPase inhibitors on the intracellular calcium activity and motility of human spermatozoa","volume":"26","author":"Williams","year":"2003","journal-title":"Int. J. Androl."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1262\/jrd.2021-143","article-title":"Involvement of Ca2+-ATPase in suppressing the appearance of bovine helically motile spermatozoa with intense force prior to cryopreservation","volume":"68","author":"Harayama","year":"2022","journal-title":"J. Reprod. Dev."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Alqu\u00e9zar-Baeta, C., Gimeno-Martos, S., Miguel-Jim\u00e9nez, S., Santolaria, P., Y\u00e1niz, J., Palac\u00edn, I., Casao, A., Cebri\u00e1n-P\u00e9rez, J.\u00c1., Mui\u00f1o-Blanco, T., and P\u00e9rez-P\u00e9, R. (2019). OpenCASA: A new open-source and scalable tool for sperm quality analysis. PLoS Comput. Biol., 15, Available online: https:\/\/pubmed.ncbi.nlm.nih.gov\/30657753\/.","DOI":"10.1371\/journal.pcbi.1006691"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1095\/biolreprod52.3.540","article-title":"Bicarbonate\/CO2 is not required for zona pellucida- or progesterone- induced acrosomal exocytosis of mouse spermatozoa but is essential for capacitation","volume":"52","author":"Shi","year":"1995","journal-title":"Biol. Reprod."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.ydbio.2016.05.008","article-title":"CaMKII prevents spontaneous acrosomal exocytosis in sperm through induction of actin polymerization","volume":"415","author":"Shabtay","year":"2016","journal-title":"Dev. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Agarwal, A., Gupta, S., and Sharma, R. (2016). Eosin-Nigrosin Staining Procedure. Andrological Evaluation of Male Infertility: A Laboratory Guide, Springer. Available online: https:\/\/link.springer.com\/chapter\/10.1007\/978-3-319-26797-5_8.","DOI":"10.1007\/978-3-319-26797-5_8"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1530\/jrf.0.0990135","article-title":"Ca(2+)-related changes in the capacitation state of human spermatozoa assessed by a chlortetracycline fluorescence assay","volume":"99","author":"DasGupta","year":"1993","journal-title":"J. Reprod. Fertil."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1002\/mrd.1080400213","article-title":"Ca(2+)-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."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1007\/s004040000112","article-title":"Evaluation of acrosome reaction and viability of human sperm with two fluorescent dyes","volume":"266","author":"Ozaki","year":"2002","journal-title":"Arch. Gynecol. Obstet."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41597-025-05177-4","article-title":"3D+t Multifocal Imaging Dataset of Human Sperm","volume":"12","author":"Montoya","year":"2025","journal-title":"Sci. Data"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Ugur, M.R., Saber Abdelrahman, A., Evans, H.C., Gilmore, A.A., Hitit, M., Arifiantini, R.I., Purwantara, B., Kaya, A., and Memili, E. (2019). Advances in Cryopreservation of Bull Sperm. Front. Vet. Sci., 6, Available online: www.frontiersin.org.","DOI":"10.3389\/fvets.2019.00268"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1530\/REP-09-0134","article-title":"Ca2+-stores in sperm: Their identities and functions","volume":"138","author":"Costello","year":"2009","journal-title":"Reproduction"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e21528","DOI":"10.1096\/fj.202100122R","article-title":"Starvation induces an increase in intracellular calcium and potentiates the progesterone-induced mouse sperm acrosome reaction","volume":"35","author":"Romarowski","year":"2021","journal-title":"FASEB J."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1673","DOI":"10.1242\/jcs.02297","article-title":"Secretory pathway Ca(2+)-ATPase (SPCA1) Ca(2)+ pumps, not SERCAs, regulate complex [Ca(2+)](i) signals in human spermatozoa","volume":"118","author":"Harper","year":"2005","journal-title":"J. Cell Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1462","DOI":"10.1359\/JBMR.050324","article-title":"Thapsigargin modulates osteoclastogenesis through the regulation of RANKL-induced signaling pathways and reactive oxygen species production","volume":"20","author":"Yip","year":"2005","journal-title":"J. Bone Miner. Res."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Ma, Z., Fan, C., Yang, Y., Di, S., Hu, W., Li, T., Zhu, Y., Han, J., Xin, Z., and Wu, G. (2016). Thapsigargin sensitizes human esophageal cancer to TRAIL-induced apoptosis via AMPK activation. Sci. Rep., 6, Available online: https:\/\/www.nature.com\/articles\/srep35196.","DOI":"10.1038\/srep35196"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.etap.2015.11.020","article-title":"Thapsigargin induces apoptosis when autophagy is inhibited in HepG2 cells and both processes are regulated by ROS-dependent pathway","volume":"41","author":"Wang","year":"2016","journal-title":"Environ. Toxicol. Pharmacol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1093\/humupd\/dmi050","article-title":"Calcium signalling in human spermatozoa: A specialized \u201ctoolkit\u201d of channels, transporters and stores","volume":"12","author":"Michelangeli","year":"2006","journal-title":"Hum. Reprod. Updat."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"ra31","DOI":"10.1126\/scisignal.2001595","article-title":"Ca2+ signaling tools acquired from prostasomes are required for progesterone-induced sperm motility","volume":"4","author":"Park","year":"2011","journal-title":"Sci. Signal."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/23\/1892\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T11:32:18Z","timestamp":1764329538000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/23\/1892"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,28]]},"references-count":41,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["cells14231892"],"URL":"https:\/\/doi.org\/10.3390\/cells14231892","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,28]]}}}