{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,21]],"date-time":"2026-06-21T12:49:09Z","timestamp":1782046149023,"version":"3.54.5"},"posted":{"date-parts":[[2024,11,6]]},"group-title":"Evolutionary Biology","reference-count":87,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,11,6]],"date-time":"2024-11-06T00:00:00Z","timestamp":1730851200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,6,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Embryo implantation is essential to pregnancy in eutherian (placental) mammals. The cellular changes that enabled invasive implantation to evolve from superficial embryo apposition, the ancestral condition still retained in marsupials, have remained unclear. We generated and compared single-cell transcriptomes from the peri-implantation and non-pregnant uterus of two rodents with invasive implantation, the mouse and the guinea pig, and the opossum, a marsupial lacking invasive implantation. Cross-species analysis of endometrial cell types and ligand-receptor signaling revealed that the opossum blastocyst-stage uterus, despite lacking fetal-maternal contact, shares key features with the eutherian window of receptivity. These include a window in which the epithelial-stromal\n                  <jats:italic>IHH<\/jats:italic>\n                  -\n                  <jats:italic>PTCH1<\/jats:italic>\n                  axis is active, but the downstream\n                  <jats:italic>BMP2<\/jats:italic>\n                  -mediated signaling that enables decidualization in eutherians is antagonized through a temporary spike in glandular\n                  <jats:italic>GREM2<\/jats:italic>\n                  . We propose that a homologous progesterone-responsive endometrial remodeling reaction preceded the evolution of eutherian and marsupial modes of implantation in their respective lineages. Complementary analysis of pre- and post-implantation embryonic transcriptomes from the same species revealed broad conservation of implantation-related functional genes in the blastocyst, as well as a remarkable similarity in ligand production between the mature trophoblast and the luminal epithelium. We suggest that this redundancy between trophectoderm and luminal epithelial signaling was an enabling factor for the evolution of invasive placentation. Together, these findings suggest that eutherian implantation evolved by maternal rewiring of the output of an ancestral window of receptivity-like epithelial-stromal signaling axis toward decidualization, stabilized epithelial loss, and invasion.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Significance Statement<\/jats:title>\n                  <jats:p>Pregnancy in placental mammals begins with implantation and invasion of the embryo, whereas marsupials retain a superficial mode of embryo apposition. How this major evolutionary transition occurred is unknown. Comparing uterine and embryonic single-cell transcriptomes between model rodents and the opossum reveals that the marsupial uterus activates an epithelial-stromal signaling axis canonically associated with receptivity to implantation. In the marsupial uterus, its outputs are opposed by inhibitory signals, whereas in placental mammals, this program promotes uterine decidual transformation and invasion. Early-embryo implantation-related gene expression, in contrast, shows little change across species. These results challenge the view that implantation evolved through the gain of invasive properties by the embryo, and reveal a common developmental foundation from which placentals evolved their novel mode of gestation.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2024.11.04.621939","type":"posted-content","created":{"date-parts":[[2024,11,7]],"date-time":"2024-11-07T01:30:23Z","timestamp":1730943023000},"source":"Crossref","is-referenced-by-count":2,"title":["Hallmarks of uterine receptivity predate placental mammals"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3161-959X","authenticated-orcid":false,"given":"Silvia","family":"Basanta","sequence":"first","affiliation":[{"name":"Department of Evolutionary Biology, University of Vienna, Vienna, Austria"},{"name":"Konrad Lorenz Institute for Evolution and Cognition Research, Klosterneuburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6249-5924","authenticated-orcid":false,"given":"Daniel J.","family":"Stadtmauer","sequence":"additional","affiliation":[{"name":"Department of Ecology & Evolutionary Biology, Yale University, New Haven, CT, USA"},{"name":"Department of Genetics, Harvard Medical School, Boston, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7137-7804","authenticated-orcid":false,"given":"Jamie D.","family":"Maziarz","sequence":"additional","affiliation":[{"name":"Department of Ecology & Evolutionary Biology, Yale University, New Haven, CT, USA"},{"name":"Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8949-7479","authenticated-orcid":false,"given":"Caitlin E.","family":"McDonough-Goldstein","sequence":"additional","affiliation":[{"name":"Department of Evolutionary Biology, University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7515-7489","authenticated-orcid":false,"given":"Alison G.","family":"Cole","sequence":"additional","affiliation":[{"name":"Department of Neuroscience and Developmental Biology, University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00fclay","family":"Dagdas","sequence":"additional","affiliation":[{"name":"Department of Evolutionary Biology, University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3097-002X","authenticated-orcid":false,"given":"G\u00fcnter P.","family":"Wagner","sequence":"additional","affiliation":[{"name":"Department of Evolutionary Biology, University of Vienna, Vienna, Austria"},{"name":"Konrad Lorenz Institute for Evolution and Cognition Research, Klosterneuburg, Austria"},{"name":"Department of Ecology & Evolutionary Biology, Yale University, New Haven, CT, USA"},{"name":"Department of Animal Science, Texas A&M University, College Station, TX, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8439-9351","authenticated-orcid":false,"given":"Mihaela","family":"Pavli\u010dev","sequence":"additional","affiliation":[{"name":"Department of Evolutionary Biology, University of Vienna, Vienna, Austria"},{"name":"Konrad Lorenz Institute for Evolution and Cognition Research, Klosterneuburg, Austria"},{"name":"Complexity Science Hub Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026062105251091000_2024.11.04.621939v2.1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod12.1.41"},{"key":"2026062105251091000_2024.11.04.621939v2.2","doi-asserted-by":"crossref","unstructured":"C. H. Tyndale-Biscoe , M. B. Renfree , Reproductive physiology of marsupials (Cambridge University Press, 1987).","DOI":"10.1017\/CBO9780511623493"},{"key":"2026062105251091000_2024.11.04.621939v2.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.jri.2019.102626"},{"key":"2026062105251091000_2024.11.04.621939v2.4","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2771"},{"key":"2026062105251091000_2024.11.04.621939v2.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.tem.2019.11.008"},{"key":"2026062105251091000_2024.11.04.621939v2.6","doi-asserted-by":"publisher","DOI":"10.1530\/ror.0.0050006"},{"key":"2026062105251091000_2024.11.04.621939v2.7","doi-asserted-by":"publisher","DOI":"10.1098\/rspb.2019.0691"},{"key":"2026062105251091000_2024.11.04.621939v2.8","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0511344103"},{"key":"2026062105251091000_2024.11.04.621939v2.9","doi-asserted-by":"publisher","DOI":"10.1002\/jez.b.21079"},{"key":"2026062105251091000_2024.11.04.621939v2.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.placenta.2009.08.004"},{"key":"2026062105251091000_2024.11.04.621939v2.11","doi-asserted-by":"crossref","unstructured":"K. Kin , J. Maziarz , G. P. Wagner , Immunohistological Study of the Endometrial Stromal Fibroblasts in the Opossum, Monodelphis domestica: Evidence for Homology with Eutherian Stromal Fibroblasts1. Biol. Reprod. 90 (2014).","DOI":"10.1095\/biolreprod.113.115139"},{"key":"2026062105251091000_2024.11.04.621939v2.12","doi-asserted-by":"publisher","DOI":"10.1093\/icb\/icaa030"},{"key":"2026062105251091000_2024.11.04.621939v2.13","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2629-6"},{"key":"2026062105251091000_2024.11.04.621939v2.14","doi-asserted-by":"publisher","DOI":"10.1159\/000016733"},{"key":"2026062105251091000_2024.11.04.621939v2.15","doi-asserted-by":"publisher","DOI":"10.1530\/jrf.0.0390173"},{"key":"2026062105251091000_2024.11.04.621939v2.16","doi-asserted-by":"publisher","DOI":"10.1007\/BF00215890"},{"key":"2026062105251091000_2024.11.04.621939v2.17","doi-asserted-by":"publisher","DOI":"10.1002\/jmor.20282"},{"key":"2026062105251091000_2024.11.04.621939v2.18","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1701129114"},{"key":"2026062105251091000_2024.11.04.621939v2.19","doi-asserted-by":"publisher","DOI":"10.1098\/rspb.2017.1694"},{"key":"2026062105251091000_2024.11.04.621939v2.20","doi-asserted-by":"publisher","DOI":"10.1016\/j.placenta.2006.11.002"},{"key":"2026062105251091000_2024.11.04.621939v2.21","doi-asserted-by":"publisher","DOI":"10.1038\/s41559-025-02748-x"},{"key":"2026062105251091000_2024.11.04.621939v2.22","doi-asserted-by":"publisher","DOI":"10.1186\/s13578-022-00749-y"},{"key":"2026062105251091000_2024.11.04.621939v2.23","doi-asserted-by":"publisher","DOI":"10.7554\/elife.66747"},{"key":"2026062105251091000_2024.11.04.621939v2.24","doi-asserted-by":"crossref","first-page":"5378","DOI":"10.1038\/s41467-019-13363-3","article-title":"Neonatal Wnt-dependent Lgr5 positive stem cells are essential for uterine gland development","volume":"10","year":"2019","journal-title":"Nat. Commun"},{"key":"2026062105251091000_2024.11.04.621939v2.25","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.100291"},{"key":"2026062105251091000_2024.11.04.621939v2.26","first-page":"1","article-title":"Histomorphological and Histochemical Study of the Ovary and the Uterine Tubes of the Adult Guinea Pigs (Cavica porcellus)","volume":"11","year":"2019","journal-title":"Egypt. Acad. J. Biol. Sci. Histol. Histochem"},{"key":"2026062105251091000_2024.11.04.621939v2.27","doi-asserted-by":"crossref","first-page":"100503","DOI":"10.1016\/j.coemr.2024.100503","article-title":"Hormonal regulation of cilia in the female reproductive tract","volume":"34","year":"2024","journal-title":"Curr. Opin. Endocr. Metab. Res"},{"key":"2026062105251091000_2024.11.04.621939v2.28","doi-asserted-by":"publisher","DOI":"10.1096\/fj.202002123R"},{"key":"2026062105251091000_2024.11.04.621939v2.29","doi-asserted-by":"publisher","DOI":"10.1002\/jez.1402660511"},{"key":"2026062105251091000_2024.11.04.621939v2.30","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-07728-1"},{"key":"2026062105251091000_2024.11.04.621939v2.31","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA119.008759"},{"key":"2026062105251091000_2024.11.04.621939v2.32","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1111\/j.1748-1716.2011.02297.x","article-title":"Wnt11 in 2011\u2013the regulation and function of a non-canonical Wnt","volume":"204","year":"2012","journal-title":"Acta Physiol"},{"key":"2026062105251091000_2024.11.04.621939v2.33","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.111.091769"},{"key":"2026062105251091000_2024.11.04.621939v2.34","doi-asserted-by":"publisher","DOI":"10.1097\/00006254-200405000-00025"},{"key":"2026062105251091000_2024.11.04.621939v2.35","doi-asserted-by":"crossref","first-page":"e3003670","DOI":"10.1371\/journal.pbio.3003670","article-title":"Fetal inflammatory signals regulate maternal investment during marsupial pregnancy","volume":"24","year":"2026","journal-title":"PLOS Biol"},{"key":"2026062105251091000_2024.11.04.621939v2.36","doi-asserted-by":"publisher","DOI":"10.1387\/ijdb.130348mb"},{"key":"2026062105251091000_2024.11.04.621939v2.37","doi-asserted-by":"publisher","DOI":"10.1038\/s41556-024-01469-w"},{"key":"2026062105251091000_2024.11.04.621939v2.38","doi-asserted-by":"publisher","DOI":"10.1038\/ng1874"},{"key":"2026062105251091000_2024.11.04.621939v2.39","doi-asserted-by":"publisher","DOI":"10.1006\/dbio.2002.0645"},{"key":"2026062105251091000_2024.11.04.621939v2.40","doi-asserted-by":"publisher","DOI":"10.1210\/me.2001-0154"},{"key":"2026062105251091000_2024.11.04.621939v2.41","doi-asserted-by":"crossref","first-page":"4421","DOI":"10.1038\/s41467-018-06652-w","article-title":"SOX17 regulates uterine epithelial\u2013stromal cross-talk acting via a distal enhancer upstream of Ihh","volume":"9","year":"2018","journal-title":"Nat. Commun"},{"key":"2026062105251091000_2024.11.04.621939v2.42","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.151180898"},{"key":"2026062105251091000_2024.11.04.621939v2.43","doi-asserted-by":"publisher","DOI":"10.1210\/en.141.12.4365"},{"key":"2026062105251091000_2024.11.04.621939v2.44","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M114.588897"},{"key":"2026062105251091000_2024.11.04.621939v2.45","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.0030102"},{"key":"2026062105251091000_2024.11.04.621939v2.46","doi-asserted-by":"publisher","DOI":"10.1210\/en.2008-1691"},{"key":"2026062105251091000_2024.11.04.621939v2.47","doi-asserted-by":"publisher","DOI":"10.1016\/j.mce.2011.10.028"},{"key":"2026062105251091000_2024.11.04.621939v2.48","doi-asserted-by":"publisher","DOI":"10.1210\/en.2012-1585"},{"key":"2026062105251091000_2024.11.04.621939v2.49","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M402376200"},{"key":"2026062105251091000_2024.11.04.621939v2.50","doi-asserted-by":"crossref","unstructured":"A. Piron , et al., RedRibbon: A new rank\u2013rank hypergeometric overlap for gene and transcript expression signatures. Life Sci. Alliance 7 (2024).","DOI":"10.26508\/lsa.202302203"},{"key":"2026062105251091000_2024.11.04.621939v2.51","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.2005594"},{"key":"2026062105251091000_2024.11.04.621939v2.52","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv134"},{"key":"2026062105251091000_2024.11.04.621939v2.53","doi-asserted-by":"publisher","DOI":"10.1186\/s12864-024-10815-z"},{"key":"2026062105251091000_2024.11.04.621939v2.54","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2016.03.023"},{"key":"2026062105251091000_2024.11.04.621939v2.55","doi-asserted-by":"publisher","DOI":"10.1023\/A:1015407012559"},{"key":"2026062105251091000_2024.11.04.621939v2.56","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1007\/s11626-020-00522-w","article-title":"Spontaneous in vitro hatching of the human blastocyst: the proteomics of initially hatching cells","volume":"56","year":"2020","journal-title":"Vitro Cell. Dev. Biol. - Anim"},{"key":"2026062105251091000_2024.11.04.621939v2.57","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-03092-4"},{"key":"2026062105251091000_2024.11.04.621939v2.58","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1016\/j.theriogenology.2023.10.011","article-title":"Effect of blastocyst development on hatching and embryo implantation","volume":"214","year":"2024","journal-title":"Theriogenology"},{"key":"2026062105251091000_2024.11.04.621939v2.59","doi-asserted-by":"publisher","DOI":"10.1210\/me.2005-0058"},{"key":"2026062105251091000_2024.11.04.621939v2.60","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsbmb.2008.12.024"},{"key":"2026062105251091000_2024.11.04.621939v2.61","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.7.3115"},{"key":"2026062105251091000_2024.11.04.621939v2.62","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod36.1.211"},{"key":"2026062105251091000_2024.11.04.621939v2.63","doi-asserted-by":"publisher","DOI":"10.7554\/elife.61257"},{"key":"2026062105251091000_2024.11.04.621939v2.64","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2014.06.027"},{"key":"2026062105251091000_2024.11.04.621939v2.65","doi-asserted-by":"publisher","DOI":"10.1086\/737357"},{"key":"2026062105251091000_2024.11.04.621939v2.66","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2016.07.046"},{"key":"2026062105251091000_2024.11.04.621939v2.67","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.00342-07"},{"key":"2026062105251091000_2024.11.04.621939v2.68","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M704723200"},{"key":"2026062105251091000_2024.11.04.621939v2.69","first-page":"227","article-title":"Impaired bone morphogenetic protein (BMP) signaling pathways disrupt decidualization in endometriosis. Commun","volume":"7","year":"2024","journal-title":"Biol"},{"key":"2026062105251091000_2024.11.04.621939v2.70","doi-asserted-by":"crossref","first-page":"bqae049","DOI":"10.1210\/endocr\/bqae049","article-title":"BMP4 in Human Endometrial Stromal Cells Can Affect Decidualization by Regulating FOXO1 Expression","volume":"165","year":"2024","journal-title":"Endocrinology"},{"key":"2026062105251091000_2024.11.04.621939v2.71","doi-asserted-by":"publisher","DOI":"10.1262\/jrd.19120"},{"key":"2026062105251091000_2024.11.04.621939v2.72","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-30194-x"},{"key":"2026062105251091000_2024.11.04.621939v2.73","doi-asserted-by":"publisher","DOI":"10.1038\/ng.917"},{"key":"2026062105251091000_2024.11.04.621939v2.74","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msaa274"},{"key":"2026062105251091000_2024.11.04.621939v2.75","doi-asserted-by":"publisher","DOI":"10.1210\/endo-95-5-1486"},{"key":"2026062105251091000_2024.11.04.621939v2.76","doi-asserted-by":"publisher","DOI":"10.1677\/joe.0.0510333"},{"key":"2026062105251091000_2024.11.04.621939v2.77","doi-asserted-by":"publisher","DOI":"10.1530\/jrf.0.0950303"},{"key":"2026062105251091000_2024.11.04.621939v2.78","doi-asserted-by":"publisher","DOI":"10.1016\/j.theriogenology.2019.06.026"},{"key":"2026062105251091000_2024.11.04.621939v2.79","doi-asserted-by":"crossref","first-page":"58","DOI":"10.3390\/mps4030058","article-title":"Time mating guinea pigs by monitoring changes to the vaginal membrane throughout the estrus cycle and with ultrasound confirmation","volume":"4","year":"2021","journal-title":"Methods Protoc"},{"key":"2026062105251091000_2024.11.04.621939v2.80","doi-asserted-by":"crossref","unstructured":"A. F. Ajayi , R. E. Akhigbe , Staging of the estrous cycle and induction of estrus in experimental rodents: an update. Fertil. Res. Pract. 6, 5 (2020).","DOI":"10.1186\/s40738-020-00074-3"},{"key":"2026062105251091000_2024.11.04.621939v2.81","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-023-01943-7"},{"key":"2026062105251091000_2024.11.04.621939v2.82","doi-asserted-by":"publisher","DOI":"10.5281\/zenodo.6349517"},{"key":"2026062105251091000_2024.11.04.621939v2.83","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-017-1382-0"},{"key":"2026062105251091000_2024.11.04.621939v2.84","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-019-0619-0"},{"key":"2026062105251091000_2024.11.04.621939v2.85","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-015-0844-5"},{"key":"2026062105251091000_2024.11.04.621939v2.86","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btac757"},{"key":"2026062105251091000_2024.11.04.621939v2.87","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-04918-4"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2024.11.04.621939","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,21]],"date-time":"2026-06-21T12:25:23Z","timestamp":1782044723000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2024.11.04.621939"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,6]]},"references-count":87,"URL":"https:\/\/doi.org\/10.1101\/2024.11.04.621939","relation":{},"subject":[],"published":{"date-parts":[[2024,11,6]]},"subtype":"preprint"}}