{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T13:31:25Z","timestamp":1791811885737,"version":"4.3.4"},"reference-count":46,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,1]],"date-time":"2025-09-01T00:00:00Z","timestamp":1756684800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CIMB"],"abstract":"<jats:p>This study aimed to describe phospholipase C zeta (PLC\u03b6) deficiency from patients who experienced oocyte fertilization failure following intracytoplasmic sperm injection (ICSI) and to investigate the relationship between sperm DNA fragmentation, chromatin decondensation, and PLC\u03b6. A total of 135 patients participated in this study\u201465 fertile men and 70 infertile patients\u2014 and semen samples were obtained to analyze concentration, motility, and morphology. PLC\u03b6 protein levels were assessed by immunofluorescence and quantitative techniques, DNA fragmentation by TUNEL essay, and chromatin decondensation by aniline blue staining. The proportion of spermatozoa presenting PLC\u03b6 was significantly lower in infertile patients (18.41 \u00b1 18.84%) compared to fertile controls (67.31 \u00b1 13.79%) (p &lt; 0.001). A significant decrease in PLC\u03b6 protein levels was observed in infertile patients compared to fertile controls, which was the same for localization patterns for each region (acrosomal, equatorial, and combination of these regions). Significant correlations were also observed between sperm parameters and PLC\u03b6 levels, DNA fragmentation, and chromatin decondensation. Furthermore, a statistically significant correlation was detected between the percentage of spermatozoa presenting PLC\u03b6 and DNA integrity (p &lt; 0.001). In summary, DNA fragmentation and chromatin decondensation are associated with alterations in the localization patterns and reduced protein levels of PLC\u03b6, which may contribute to total fertilization failure.<\/jats:p>","DOI":"10.3390\/cimb47090707","type":"journal-article","created":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T13:01:13Z","timestamp":1756818073000},"page":"707","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Effect of Sperm DNA Fragmentation and Chromatin Decondensation on PLC\u03b6 Efficacy in Infertile Patients"],"prefix":"10.3390","volume":"47","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9303-8566","authenticated-orcid":false,"given":"Soukaina","family":"Azil","sequence":"first","affiliation":[{"name":"Immunopathology-Immunotherapy-Immunomonitoring Laboratory, Faculty of Medicine, Mohammed VI University of Sciences and Health, Casablanca 82403, Morocco"},{"name":"In Vitro Fertilization Center IRIFIV, Iris Clinic, Laboratory of Medical Analysis and Reproductive Biology, Labomac, Casablanca 20100, Morocco"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ismail","family":"Kaarouch","sequence":"additional","affiliation":[{"name":"African Fertility Clinic, Private Clinic of Human Reproduction and Endoscopic Surgery, Casablanca 20000, Morocco"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9776-7459","authenticated-orcid":false,"given":"Debbie","family":"Montjean","sequence":"additional","affiliation":[{"name":"Fertilys Fertility Centers Laval and Brossard, 1950 Maurice-Gauvin Street, Laval, QC H7S 1Z5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie-H\u00e9l\u00e8ne Godin","family":"Pag\u00e9","sequence":"additional","affiliation":[{"name":"Fertilys Fertility Centers Laval and Brossard, 1950 Maurice-Gauvin Street, Laval, QC H7S 1Z5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosalie","family":"Cabry","sequence":"additional","affiliation":[{"name":"Reproductive Medicine, Reproductive Biology and Genetics, Peritox Laboratory, University Hospital and School of Medicine, Picardie University Jules Verne, 80054 Amiens, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Noureddine","family":"Louanjli","sequence":"additional","affiliation":[{"name":"In Vitro Fertilization Center IRIFIV, Iris Clinic, Laboratory of Medical Analysis and Reproductive Biology, Labomac, Casablanca 20100, Morocco"},{"name":"African Fertility Clinic, Private Clinic of Human Reproduction and Endoscopic Surgery, Casablanca 20000, Morocco"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bouchra","family":"Ghazi","sequence":"additional","affiliation":[{"name":"Immunopathology-Immunotherapy-Immunomonitoring Laboratory, Faculty of Medicine, Mohammed VI University of Sciences and Health, Casablanca 82403, Morocco"},{"name":"Mohammed VI International University Hospital, Bouskoura 27182, Morocco"},{"name":"Mohammed VI Center for Research and Innovation (CM6RI), Rabat 10112, Morocco"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moncef","family":"Benkhalifa","sequence":"additional","affiliation":[{"name":"Reproductive Medicine, Reproductive Biology and Genetics, Peritox Laboratory, University Hospital and School of Medicine, Picardie University Jules Verne, 80054 Amiens, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1273","DOI":"10.1007\/s10815-020-01748-z","article-title":"Increasing Associations between Defects in Phospholipase C Zeta and Conditions of Male Infertility: Not Just ICSI Failure?","volume":"37","author":"Kashir","year":"2020","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1016\/j.jbior.2015.11.009","article-title":"Phospholipase C Zeta (PLC\u03b6) and Male Infertility: Clinical Update and Topical Developments","volume":"61","author":"Amdani","year":"2016","journal-title":"Adv. Biol. Regul."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Saleh, A., Kashir, J., Thanassoulas, A., Safieh-Garabedian, B., Lai, F.A., and Nomikos, M. (2020). Essential Role of Sperm-Specific PLC-Zeta in Egg Activation and Male Factor Infertility: An Update. Front. Cell Dev. Biol., 8.","DOI":"10.3389\/fcell.2020.00028"},{"key":"ref_4","first-page":"467","article-title":"Phospholipase C Zeta: A Hidden Face of Sperm for Oocyte Activation and Early Embryonic Development","volume":"67","author":"Azil","year":"2024","journal-title":"Korean J. Obstet. Gynecol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"hoac003","DOI":"10.1093\/hropen\/hoac003","article-title":"Oocyte Activation Deficiency and Assisted Oocyte Activation: Mechanisms, Obstacles and Prospects for Clinical Application","volume":"2022","author":"Kashir","year":"2022","journal-title":"Hum. Reprod. Open"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ribas-Maynou, J., Llavanera, M., Mateo-Otero, Y., Garcia-Bonavila, E., Delgado-Berm\u00fadez, A., and Yeste, M. (2021). Direct but Not Indirect Methods Correlate the Percentages of Sperm with Altered Chromatin to the Intensity of Chromatin Damage. Front. Vet. Sci., 8.","DOI":"10.3389\/fvets.2021.719319"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.theriogenology.2021.08.024","article-title":"The TUNEL Assay Underestimates the Incidence of DNA Damage in Pig Sperm Due to Chromatin Condensation","volume":"174","author":"Bonet","year":"2021","journal-title":"Theriogenology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2489","DOI":"10.1007\/s43032-023-01209-9","article-title":"Advanced Paternal Age Affects the Sperm DNA Fragmentation Index and May Lead to Lower Good-Quality Blastocysts","volume":"30","author":"Donatti","year":"2023","journal-title":"Reprod. Sci."},{"key":"ref_9","first-page":"588","article-title":"Relationship between Potential Sperm Factors Involved in Oocyte Activation and Sperm DNA Fragmentation with Intra-Cytoplasmic Sperm Injection Clinical Outcomes","volume":"18","author":"Tavalaee","year":"2016","journal-title":"Cell J."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1080\/19396368.2017.1298006","article-title":"Relationship between Phospholipase C-Zeta, Semen Parameters, and Chromatin Status","volume":"63","author":"Tavalaee","year":"2017","journal-title":"Syst. Biol. Reprod. Med."},{"key":"ref_11","unstructured":"Bj\u00f6rndahl, L., Apolikhin, O., Baldi, E., Barratt, C.L.R., Festin, M.P.R., Kirkman-Brown, J.C., and Wang, C. (2021). WHO Laboratory Manual for the Examination and Processing of Human Semen, World Health Organization."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e13738","DOI":"10.1111\/and.13738","article-title":"TUNEL Assay\u2014Standardized Method for Testing Sperm DNA Fragmentation","volume":"53","author":"Sharma","year":"2021","journal-title":"Andrologia"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"578631","DOI":"10.1155\/2013\/578631","article-title":"Assessment of Chromatin Maturity in Human Spermatozoa: Useful Aniline Blue Assay for Routine Diagnosis of Male Infertility","volume":"2013","author":"Sellami","year":"2013","journal-title":"Adv. Urol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2513","DOI":"10.1093\/humrep\/den280","article-title":"The Pattern of Localization of the Putative Oocyte Activation Factor, Phospholipase C\u03b6, in Uncapacitated, Capacitated, and Ionophore-Treated Human Spermatozoa","volume":"23","author":"Grasa","year":"2008","journal-title":"Hum. Reprod."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1016\/j.fertnstert.2015.05.018","article-title":"Total Levels, Localization Patterns, and Proportions of Sperm Exhibiting Phospholipase C Zeta Are Significantly Correlated with Fertilization Rates after Intracytoplasmic Sperm Injection","volume":"104","author":"Yelumalai","year":"2015","journal-title":"Fertil. Steril."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1016\/j.fertnstert.2004.04.033","article-title":"Relationship among Standard Semen Parameters, Glutathione Peroxidase\/Glutathione Reductase Activity, and MRNA Expression and Reduced Glutathione Content in Ejaculated Spermatozoa from Fertile and Infertile Men","volume":"82","author":"Garrido","year":"2004","journal-title":"Fertil. Steril."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.jbior.2017.10.012","article-title":"Phospholipase C Zeta and Calcium Oscillations at Fertilisation: The Evidence, Applications, and Further Questions","volume":"67","author":"Kashir","year":"2018","journal-title":"Adv. Biol. Regul."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Kaltsas, A., Markou, E., Kyrgiafini, M.-A., Zikopoulos, A., Symeonidis, E.N., Dimitriadis, F., Zachariou, A., Sofikitis, N., and Chrisofos, M. (2025). Oxidative-Stress-Mediated Epigenetic Dysregulation in Spermatogenesis: Implications for Male Infertility and Offspring Health. Genes, 16.","DOI":"10.3390\/genes16010093"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"104","DOI":"10.5534\/wjmh.200195","article-title":"Advanced Paternal Age and Sperm DNA Fragmentation: A Systematic Review","volume":"40","author":"Gonzalez","year":"2021","journal-title":"World J. Mens. Health"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Kaltsas, A., Moustakli, E., Zikopoulos, A., Georgiou, I., Dimitriadis, F., Symeonidis, E.N., Markou, E., Michaelidis, T.M., Tien, D.M.B., and Giannakis, I. (2023). Impact of Advanced Paternal Age on Fertility and Risks of Genetic Disorders in Offspring. Genes, 14.","DOI":"10.3390\/genes14020486"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1256","DOI":"10.1093\/humrep\/deae078","article-title":"The Mammalian Sperm Factor Phospholipase C Zeta Is Critical for Early Embryo Division and Pregnancy in Humans and Mice","volume":"39","author":"Kashir","year":"2024","journal-title":"Hum. Reprod."},{"key":"ref_22","first-page":"38","article-title":"Evaluation of PAWP and PLC\u03b6 Expression in Infertile Men with Previous ICSI Fertilization Failure","volume":"15","author":"Azad","year":"2018","journal-title":"Urol. J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1002\/mrd.23293","article-title":"The Alteration of PLC\u03b6 Protein Expression in Unexplained Infertile and Asthenoteratozoospermic Patients: A Potential Effect on Sperm Fertilization Ability","volume":"87","author":"Rahimizadeh","year":"2020","journal-title":"Mol. Reprod. Dev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1093\/humrep\/dez282","article-title":"Novel Mutations in PLCZ1 Cause Male Infertility Due to Fertilization Failure or Poor Fertilization","volume":"35","author":"Yan","year":"2020","journal-title":"Hum. Reprod."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2417","DOI":"10.1093\/humrep\/dep207","article-title":"Reduced Amounts and Abnormal Forms of Phospholipase C Zeta (PLC\u03b6) in Spermatozoa from Infertile Men","volume":"24","author":"Heytens","year":"2009","journal-title":"Hum. Reprod."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1111\/cge.13636","article-title":"Novel Homozygous Variations in PLCZ1 Lead to Poor or Failed Fertilization Characterized by Abnormal Localization Patterns of PLC\u03b6 in Sperm","volume":"97","author":"Dai","year":"2020","journal-title":"Clin. Genet."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Palermo, G.D., Neri, Q.V., Schlegel, P.N., and Rosenwaks, Z. (2014). Intracytoplasmic Sperm Injection (ICSI) in Extreme Cases of Male Infertility. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0113671"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1016\/j.fertnstert.2009.03.061","article-title":"Artificial Oocyte Activation and Intracytoplasmic Sperm Injection","volume":"94","author":"Deemeh","year":"2010","journal-title":"Fertil. Steril."},{"key":"ref_29","first-page":"e14269","article-title":"Phospholipase C-zeta Levels Are Not Correlated with Fertilisation Rates in Infertile Couples","volume":"54","author":"Cakar","year":"2022","journal-title":"Andrologia"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Kashir, J., Jones, C., and Coward, K. (2014). Oocyte Activation and Phospholipase C Zeta (PLC\u03b6): Male Infertility and Implications for Therapeutic Intervention. Phospholipases Health Dis., 263\u2013281.","DOI":"10.1007\/978-1-4939-0464-8_16"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1007\/s10815-020-01719-4","article-title":"A Homozygous Nonsense Mutation of PLCZ1 Cause Male Infertility with Oocyte Activation Deficiency","volume":"37","author":"Wang","year":"2020","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"878","DOI":"10.1093\/hmg\/ddv617","article-title":"Homozygous Mutation of PLCZ1 Leads to Defective Human Oocyte Activation and Infertility That Is Not Rescued by the WW-Binding Protein PAWP","volume":"25","author":"Escoffier","year":"2016","journal-title":"Hum. Mol. Genet."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1007\/s10815-016-0718-0","article-title":"PLC\u03b6 Sequence, Protein Levels, and Distribution in Human Sperm Do Not Correlate with Semen Characteristics and Fertilization Rates after ICSI","volume":"33","author":"Barragan","year":"2016","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1038\/nature11396","article-title":"Rate of de Novo Mutations and the Importance of Father\u2019s Age to Disease Risk","volume":"488","author":"Kong","year":"2012","journal-title":"Nature"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"232","DOI":"10.5468\/ogs.2015.58.3.232","article-title":"Relationship between Phospholipase C Zeta Immunoreactivity and DNA Fragmentation and Oxidation in Human Sperm","volume":"58","author":"Park","year":"2015","journal-title":"Obs. Gynecol. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Solovova, O.A., and Chernykh, V.B. (2022). Genetics of Oocyte Maturation Defects and Early Embryo Development Arrest. Genes, 13.","DOI":"10.3390\/genes13111920"},{"key":"ref_37","first-page":"289","article-title":"Biological Basis for Human Capacitation\u2014Revisited","volume":"23","year":"2017","journal-title":"Hum. Reprod. Update"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1007\/s10815-018-1342-y","article-title":"Altered Three-Dimensional Organization of Sperm Genome in DPY19L2-Deficient Globozoospermic Patients","volume":"36","author":"Abdelhedi","year":"2019","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Halvaei, I., Litzky, J., and Esfandiari, N. (2020). Advanced Paternal Age: Effects on Sperm Parameters, Assisted Reproduction Outcomes and Offspring Health. Reprod. Biol. Endocrinol., 18.","DOI":"10.1186\/s12958-020-00668-y"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Chan, P.T.K., and Robaire, B. (2022). Advanced Paternal Age and Future Generations. Front. Endocrinol., 13.","DOI":"10.3389\/fendo.2022.897101"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1109","DOI":"10.1007\/s11154-022-09759-0","article-title":"Reproductive Axis Ageing and Fertility in Men","volume":"23","author":"Anderson","year":"2022","journal-title":"Rev. Endocr. Metab. Disord."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"178","DOI":"10.4103\/aja.aja_67_20","article-title":"Advancing Male Age Differentially Alters Levels and Localization Patterns of PLCzeta in Sperm and Testes from Different Mouse Strains","volume":"23","author":"Kashir","year":"2021","journal-title":"Asian J. Androl."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Sugita, H., Takarabe, S., Kageyama, A., Kawata, Y., and Ito, J. (2024). Molecular Mechanism of Oocyte Activation in Mammals: Past, Present, and Future Directions. Biomolecules, 14.","DOI":"10.3390\/biom14030359"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Saleh, A., Thanassoulas, A., Aliyev, E., Swann, K., Naija, A., Yalcin, H.C., Lai, F.A., and Nomikos, M. (2024). Development of Recombinant PLC-Zeta Protein as a Therapeutic Intervention for the Clinical Treatment of Oocyte Activation Failure. Biomedicines, 12.","DOI":"10.3390\/biomedicines12061183"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1093\/molehr\/gay038","article-title":"Mammalian Sperm Contain Two Factors for Calcium Release and Egg Activation: Phospholipase C Zeta and a Cryptic Activating Factor","volume":"24","author":"Jones","year":"2018","journal-title":"Mol. Hum. Reprod."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"F53","DOI":"10.1530\/REP-21-0458","article-title":"Sperm Factors and Egg Activation: Phospholipase C Zeta (PLCZ1) and the Clinical Diagnosis of Oocyte Activation Deficiency","volume":"164","author":"Jones","year":"2022","journal-title":"Reproduction"}],"container-title":["Current Issues in Molecular Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1467-3045\/47\/9\/707\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:37:08Z","timestamp":1760035028000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1467-3045\/47\/9\/707"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,1]]},"references-count":46,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["cimb47090707"],"URL":"https:\/\/doi.org\/10.3390\/cimb47090707","relation":{},"ISSN":["1467-3045"],"issn-type":[{"value":"1467-3045","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,1]]}}}