{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T02:04:21Z","timestamp":1779761061979,"version":"3.53.1"},"posted":{"date-parts":[[2026,5,22]]},"group-title":"Cell Biology","reference-count":62,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T00:00:00Z","timestamp":1779408000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"crossref","award":["GM135549"],"award-info":[{"award-number":["GM135549"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development","award":["HD082375"],"award-info":[{"award-number":["HD082375"]}]},{"name":"Cancer Center of Illinois","award":["Seed Grant"],"award-info":[{"award-number":["Seed Grant"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,22]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Phthalates are pervasive endocrine-disrupting chemicals widely used in consumer products. The wide use of many phthalates results in chronic human exposure to complex mixtures rather than single compounds. Despite extensive studies on individual compounds, the combined effects of phthalate metabolites on oogenesis remain poorly understood. Here, we developed a precise microinjection-based single-oocyte toxicological assay to examine the impact of a defined phthalate metabolite mixture on meiotic progression. Phthalate mixture exposure markedly impaired oocyte maturation, as most oocytes failed to extrude the first polar body. Mechanistic analyses revealed severe meiotic defects, including disrupted spindle morphology, chromosome misalignment, disorganized actin cytoskeleton, and impaired mitochondrial function, accompanied by excessive reactive oxygen species (ROS) accumulation and DNA damage. Single-cell transcriptomic profiling further identified differentially expressed genes enriched in biological processes related to exocytosis, secretory pathway regulation, and cytoskeletal organization, as well as in MAPK, JAK-STAT, cGMP-PKG, and GnRH signaling pathways that are essential for follicular development and oocyte maturation. Together, these findings demonstrate that combined phthalate exposure directly compromises female gamete quality and underscore the importance of evaluating mixture effects when assessing risks to women\u2019s reproductive health.<\/jats:p>","DOI":"10.64898\/2026.05.20.726577","type":"posted-content","created":{"date-parts":[[2026,5,23]],"date-time":"2026-05-23T00:20:15Z","timestamp":1779495615000},"source":"Crossref","is-referenced-by-count":0,"title":["Effects of Phthalate Metabolite Mixture Exposure on Mouse Oocyte Development"],"prefix":"10.64898","author":[{"given":"Juan","family":"Dong","sequence":"first","affiliation":[{"name":"Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China"},{"name":"Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vidhi","family":"Patel","sequence":"additional","affiliation":[{"name":"Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuangqi","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hasanur","family":"Alam","sequence":"additional","affiliation":[{"name":"Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjie","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anika","family":"Roy","sequence":"additional","affiliation":[{"name":"Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leyi","family":"Wang","sequence":"additional","affiliation":[{"name":"Veterinary Diagnostic Laboratory, Department of Veterinary Clinical Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jodi A.","family":"Flaws","sequence":"additional","affiliation":[{"name":"Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA"},{"name":"Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Urbana, Illinois, 61801, USA"},{"name":"Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0966-8077","authenticated-orcid":false,"given":"Huanyu","family":"Qiao","sequence":"additional","affiliation":[{"name":"Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA"},{"name":"Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Urbana, Illinois, 61801, USA"},{"name":"Department of Molecular & Integrative Physiology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA"},{"name":"Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026052518550440000_2026.05.20.726577v1.1","unstructured":"Organization, W.H ., Infertility prevalence estimates, 1990\u20132021. 2023: World Health Organization."},{"key":"2026052518550440000_2026.05.20.726577v1.2","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00032.2022"},{"key":"2026052518550440000_2026.05.20.726577v1.3","doi-asserted-by":"publisher","DOI":"10.1093\/molehr\/gan069"},{"issue":"1","key":"2026052518550440000_2026.05.20.726577v1.4","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s12958-018-0445-8","article-title":"The art of oocyte meiotic arrest regulation","volume":"17","year":"2019","journal-title":"Reprod Biol Endocrinol"},{"issue":"5","key":"2026052518550440000_2026.05.20.726577v1.5","doi-asserted-by":"crossref","first-page":"878","DOI":"10.1093\/biolre\/ioz143","article-title":"Cyclins regulating oocyte meiotic cell cycle progressiondagger","volume":"101","year":"2019","journal-title":"Biol Reprod"},{"issue":"2","key":"2026052518550440000_2026.05.20.726577v1.6","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1042\/EBC20190068","article-title":"Advances and surprises in a decade of oocyte meiosis research","volume":"64","year":"2020","journal-title":"Essays Biochem"},{"issue":"8","key":"2026052518550440000_2026.05.20.726577v1.7","doi-asserted-by":"crossref","first-page":"5530","DOI":"10.1002\/jcp.26467","article-title":"Journey of oocyte from metaphase-I to metaphase-II stage in mammals","volume":"233","year":"2018","journal-title":"J Cell Physiol"},{"key":"2026052518550440000_2026.05.20.726577v1.8","doi-asserted-by":"publisher","DOI":"10.1093\/biolre\/ioab002"},{"key":"2026052518550440000_2026.05.20.726577v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijheh.2007.07.011"},{"key":"2026052518550440000_2026.05.20.726577v1.10","doi-asserted-by":"publisher","DOI":"10.1021\/es4042034"},{"key":"2026052518550440000_2026.05.20.726577v1.11","doi-asserted-by":"publisher","DOI":"10.1002\/mnfr.200600243"},{"key":"2026052518550440000_2026.05.20.726577v1.12","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2008.0208"},{"key":"2026052518550440000_2026.05.20.726577v1.13","doi-asserted-by":"publisher","DOI":"10.1289\/ehp.11610"},{"key":"2026052518550440000_2026.05.20.726577v1.14","doi-asserted-by":"publisher","DOI":"10.1289\/ehp.116-a334"},{"key":"2026052518550440000_2026.05.20.726577v1.15","doi-asserted-by":"publisher","DOI":"10.1016\/j.reprotox.2016.04.005"},{"key":"2026052518550440000_2026.05.20.726577v1.16","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.clinms.2020.10.002","article-title":"Development and validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous determination of phthalates and bisphenol a in serum, urine and follicular fluid","volume":"18","year":"2020","journal-title":"Clin Mass Spectrom"},{"key":"2026052518550440000_2026.05.20.726577v1.17","doi-asserted-by":"publisher","DOI":"10.1016\/j.envint.2015.08.005"},{"key":"2026052518550440000_2026.05.20.726577v1.18","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.115.129148"},{"key":"2026052518550440000_2026.05.20.726577v1.19","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.112.102467"},{"key":"2026052518550440000_2026.05.20.726577v1.20","doi-asserted-by":"publisher","DOI":"10.1016\/j.taap.2015.02.010"},{"key":"2026052518550440000_2026.05.20.726577v1.21","doi-asserted-by":"publisher","DOI":"10.1210\/er.2015-1010"},{"key":"2026052518550440000_2026.05.20.726577v1.22","doi-asserted-by":"publisher","DOI":"10.1002\/mrd.23652"},{"key":"2026052518550440000_2026.05.20.726577v1.23","first-page":"8","article-title":"The effects of phthalates on the ovary","volume":"6","year":"2015","journal-title":"Front Endocrinol (Lausanne"},{"issue":"1","key":"2026052518550440000_2026.05.20.726577v1.24","doi-asserted-by":"crossref","first-page":"e1008529","DOI":"10.1371\/journal.pgen.1008529","article-title":"Environmentally-relevant exposure to diethylhexyl phthalate (DEHP) alters regulation of double-strand break formation and crossover designation leading to germline dysfunction in Caenorhabditis elegans","volume":"16","year":"2020","journal-title":"PLoS Genet"},{"key":"2026052518550440000_2026.05.20.726577v1.25","doi-asserted-by":"publisher","DOI":"10.1016\/j.taap.2011.11.008"},{"key":"2026052518550440000_2026.05.20.726577v1.26","doi-asserted-by":"crossref","first-page":"116088","DOI":"10.1016\/j.envpol.2020.116088","article-title":"Di (2-ethylhexyl) phthalate impairs primordial follicle assembly by increasing PDE3A expression in oocytes","volume":"270","year":"2021","journal-title":"Environ Pollut"},{"issue":"A","key":"2026052518550440000_2026.05.20.726577v1.27","first-page":"388","article-title":"Dibutyl phthalate exposure disrupts the progression of meiotic prophase I by interfering with homologous recombination in fetal mouse oocytes","volume":"252","year":"2019","journal-title":"Environ Pollut"},{"issue":"1","key":"2026052518550440000_2026.05.20.726577v1.28","first-page":"217","article-title":"Effects of an Environmentally Relevant Phthalate Mixture on Cultured Mouse Antral Follicles","volume":"156","year":"2017","journal-title":"Toxicol Sci"},{"issue":"2","key":"2026052518550440000_2026.05.20.726577v1.29","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1093\/toxsci\/kfaa170","article-title":"Ovulation is Inhibited by an Environmentally Relevant Phthalate Mixture in Mouse Antral Follicles In Vitro","volume":"179","year":"2021","journal-title":"Toxicol Sci"},{"key":"2026052518550440000_2026.05.20.726577v1.30","doi-asserted-by":"crossref","first-page":"117372","DOI":"10.1016\/j.taap.2025.117372","article-title":"Neonatal exposure to an environmentally relevant phthalate mixture alters ovarian function in mice","volume":"500","year":"2025","journal-title":"Toxicol Appl Pharmacol"},{"key":"2026052518550440000_2026.05.20.726577v1.31","doi-asserted-by":"publisher","DOI":"10.1093\/biolre\/ioab010"},{"issue":"12","key":"2026052518550440000_2026.05.20.726577v1.32","doi-asserted-by":"crossref","first-page":"127019","DOI":"10.1289\/EHP11998","article-title":"Associations between Phthalate Metabolite Concentrations in Follicular Fluid and Reproductive Outcomes among Women Undergoing in Vitro Fertilization\/Intracytoplasmic Sperm Injection Treatment","volume":"131","year":"2023","journal-title":"Environ Health Perspect"},{"key":"2026052518550440000_2026.05.20.726577v1.33","doi-asserted-by":"crossref","first-page":"108383","DOI":"10.1016\/j.envint.2023.108383","article-title":"Ovarian follicular fluid levels of phthalates and benzophenones in relation to fertility outcomes","volume":"183","year":"2024","journal-title":"Environ Int"},{"key":"2026052518550440000_2026.05.20.726577v1.34","doi-asserted-by":"crossref","first-page":"108868","DOI":"10.1016\/j.reprotox.2025.108868","article-title":"Associations between urinary and follicular fluid concentrations of phthalate metabolites and reproductive outcomes in Brazilian women undergoing fertility treatment","volume":"133","year":"2025","journal-title":"Reprod Toxicol"},{"key":"2026052518550440000_2026.05.20.726577v1.35","doi-asserted-by":"publisher","DOI":"10.1007\/s10815-024-03101-0"},{"issue":"5","key":"2026052518550440000_2026.05.20.726577v1.36","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1038\/s41370-018-0050-0","article-title":"A possible approach to improving the reproducibility of urinary concentrations of phthalate metabolites and phenols during pregnancy","volume":"28","year":"2018","journal-title":"J Expo Sci Environ Epidemiol"},{"issue":"2","key":"2026052518550440000_2026.05.20.726577v1.37","doi-asserted-by":"crossref","first-page":"21","DOI":"10.3390\/toxics7020021","article-title":"A review of biomonitoring of phthalate exposures","volume":"7","year":"2019","journal-title":"Toxics"},{"issue":"4","key":"2026052518550440000_2026.05.20.726577v1.38","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1038\/s41370-023-00614-1","article-title":"Year-to-year variation in phthalate metabolites in the Midlife Women\u2019s Health Study","volume":"34","year":"2024","journal-title":"Journal of exposure science & environmental epidemiology"},{"key":"2026052518550440000_2026.05.20.726577v1.39","doi-asserted-by":"crossref","first-page":"114101","DOI":"10.1016\/j.ijheh.2022.114101","article-title":"Current exposure to phthalates and DINCH in European children and adolescents\u2013Results from the HBM4EU Aligned Studies 2014 to 2021","volume":"249","year":"2023","journal-title":"International Journal of Hygiene and Environmental Health"},{"key":"2026052518550440000_2026.05.20.726577v1.40","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2008.0208"},{"issue":"1","key":"2026052518550440000_2026.05.20.726577v1.41","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.xfss.2024.10.009","article-title":"Phthalates are detected in the follicular fluid of adolescents and oocyte donors with associated changes in the cumulus cell transcriptome","volume":"6","year":"2025","journal-title":"F&S Science"},{"key":"2026052518550440000_2026.05.20.726577v1.42","doi-asserted-by":"publisher","DOI":"10.1016\/j.taap.2019.114875"},{"issue":"3","key":"2026052518550440000_2026.05.20.726577v1.43","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1093\/biolre\/ioad178","article-title":"The role of the aryl hydrocarbon receptor in mediating the effects of mono(2-ethylhexyl) phthalate in mouse ovarian antral folliclesdagger","volume":"110","year":"2024","journal-title":"Biol Reprod"},{"key":"2026052518550440000_2026.05.20.726577v1.44","doi-asserted-by":"crossref","first-page":"109295","DOI":"10.1016\/j.envres.2020.109295","article-title":"Predictors of phthalate metabolites in urine and follicular fluid and correlations between urine and follicular fluid phthalate metabolite concentrations among women undergoing in vitro fertilization","volume":"184","year":"2020","journal-title":"Environ Res"},{"key":"2026052518550440000_2026.05.20.726577v1.45","doi-asserted-by":"crossref","first-page":"115601","DOI":"10.1016\/j.envpol.2020.115601","article-title":"Iodoacetic acid disrupts mouse oocyte maturation by inducing oxidative stress and spindle abnormalities","volume":"268","year":"2021","journal-title":"Environmental Pollution"},{"key":"2026052518550440000_2026.05.20.726577v1.46","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0497-0"},{"issue":"8","key":"2026052518550440000_2026.05.20.726577v1.47","doi-asserted-by":"crossref","first-page":"e2966","DOI":"10.1038\/cddis.2017.350","article-title":"Di (2-ethylhexyl) phthalate exposure impairs meiotic progression and DNA damage repair in fetal mouse oocytes in vitro","volume":"8","year":"2017","journal-title":"Cell Death Dis"},{"key":"2026052518550440000_2026.05.20.726577v1.48","doi-asserted-by":"crossref","first-page":"114768","DOI":"10.1016\/j.ecoenv.2023.114768","article-title":"Exposure to diisononyl phthalate deteriorates the quality of porcine oocytes by inducing the apoptosis","volume":"254","year":"2023","journal-title":"Ecotoxicol Environ Saf"},{"key":"2026052518550440000_2026.05.20.726577v1.49","doi-asserted-by":"crossref","first-page":"129740","DOI":"10.1016\/j.chemosphere.2021.129740","article-title":"Exposure to phthalates DEHP and DINP May lead to oxidative damage and lipidomic disruptions in mouse kidney","volume":"271","year":"2021","journal-title":"Chemosphere"},{"issue":"5","key":"2026052518550440000_2026.05.20.726577v1.50","doi-asserted-by":"crossref","first-page":"678","DOI":"10.3390\/antiox10050678","article-title":"Antioxidant synergy of mitochondrial phospholipase PNPLA8\/iPLA2\u03b3 with fatty acid\u2013conducting SLC25 gene family transporters","volume":"10","year":"2021","journal-title":"Antioxidants"},{"key":"2026052518550440000_2026.05.20.726577v1.51","first-page":"8","article-title":"The effects of phthalates on the ovary","volume":"6","year":"2015","journal-title":"Frontiers in endocrinology"},{"issue":"2","key":"2026052518550440000_2026.05.20.726577v1.52","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1002\/biof.5520330207","article-title":"Effect of mono-(2-ethylhexyl) phthalate (MEHP) on resumption of meiosis, in vitro maturation and embryo development of immature mouse oocytes","volume":"33","year":"2008","journal-title":"Biofactors"},{"issue":"7","key":"2026052518550440000_2026.05.20.726577v1.53","doi-asserted-by":"crossref","first-page":"e0130896","DOI":"10.1371\/journal.pone.0130896","article-title":"Carryover Effects of Acute DEHP Exposure on Ovarian Function and Oocyte Developmental Competence in Lactating Cows","volume":"10","year":"2015","journal-title":"PLoS One"},{"key":"2026052518550440000_2026.05.20.726577v1.54","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.jhazmat.2015.05.052","article-title":"DEHP exposure impairs mouse oocyte cyst breakdown and primordial follicle assembly through estrogen receptor-dependent and independent mechanisms","volume":"298","year":"2015","journal-title":"J Hazard Mater"},{"key":"2026052518550440000_2026.05.20.726577v1.55","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1016\/j.envpol.2018.12.077","article-title":"Di(n-butyl) phthalate exposure impairs meiotic competence and development of mouse oocyte","volume":"246","year":"2019","journal-title":"Environ Pollut"},{"key":"2026052518550440000_2026.05.20.726577v1.56","doi-asserted-by":"crossref","first-page":"119756","DOI":"10.1016\/j.envpol.2022.119756","article-title":"Glycine ameliorates MBP-induced meiotic abnormalities and apoptosis by regulating mitochondrial-endoplasmic reticulum interactions in porcine oocytes","volume":"309","year":"2022","journal-title":"Environ Pollut"},{"key":"2026052518550440000_2026.05.20.726577v1.57","doi-asserted-by":"publisher","DOI":"10.1016\/j.envres.2011.03.005"},{"key":"2026052518550440000_2026.05.20.726577v1.58","doi-asserted-by":"crossref","first-page":"109231","DOI":"10.1016\/j.envres.2020.109231","article-title":"Use of stored serum in the study of time trends and geographical differences in exposure of pregnant women to phthalates","volume":"184","year":"2020","journal-title":"Environ Res"},{"key":"2026052518550440000_2026.05.20.726577v1.59","doi-asserted-by":"publisher","DOI":"10.1007\/s10815-012-9775-1"},{"key":"2026052518550440000_2026.05.20.726577v1.60","doi-asserted-by":"crossref","unstructured":"Sun, X. , et al., Smart RNA Sequencing Reveals the Toxicological Effects of Diisobutyl Phthalate (DiBP) in Porcine Oocytes. Environ Sci Technol, 2024.","DOI":"10.1021\/acs.est.4c05462"},{"key":"2026052518550440000_2026.05.20.726577v1.61","doi-asserted-by":"publisher","DOI":"10.1093\/toxsci\/kft102"},{"key":"2026052518550440000_2026.05.20.726577v1.62","doi-asserted-by":"publisher","DOI":"10.1631\/jzus.B0820379"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.05.20.726577","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T01:55:21Z","timestamp":1779760521000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.05.20.726577"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,22]]},"references-count":62,"URL":"https:\/\/doi.org\/10.64898\/2026.05.20.726577","relation":{},"subject":[],"published":{"date-parts":[[2026,5,22]]},"subtype":"preprint"}}