{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T08:23:09Z","timestamp":1791361389666,"version":"4.3.1"},"posted":{"date-parts":[[2025,12,8]]},"group-title":"Developmental Biology","reference-count":64,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T00:00:00Z","timestamp":1765152000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development","award":["R01HD109232"],"award-info":[{"award-number":["R01HD109232"]}]},{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development","award":["R01HD109232-S1"],"award-info":[{"award-number":["R01HD109232-S1"]}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"crossref","award":["R35GM142875"],"award-info":[{"award-number":["R35GM142875"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,12,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Cilia are microtubule-based organelles essential for signaling and motility, and their dysfunction causes ciliopathies often associated with infertility. In male germ cells, two types of cilia are present: zygotene primary cilia and sperm flagella. To define the role of cilia in spermatogenesis, we conditionally ablated the distal appendage protein CEP164, required for basal body docking and ciliogenesis, in male germ cells. CEP164 localized to the mother centriole\/basal body throughout spermatogenesis, and its loss led to male infertility accompanied by absence of both zygotene cilia and sperm flagella. Despite defective ciliogenesis, meiotic chromosome pairing and DNA double-strand break repair proceeded normally. However, round spermatids exhibited basal body docking and flagellogenesis defects, and frequently formed supernumerary centriole clusters that were subsequently eliminated via residual bodies. Live-cell imaging revealed that centrioles were highly mobile, and centriole pairs from neighboring cells often associated through intercellular bridges, forming aggregates. These results establish that basal body docking is crucial for retaining centrioles within spermatids, and its disruption leads to centriole clustering and loss. In contrast, zygotene cilia are dispensable for meiotic chromosome pairing and DNA repair during mammalian spermatogenesis.<\/jats:p>","DOI":"10.64898\/2025.12.04.692363","type":"posted-content","created":{"date-parts":[[2025,12,9]],"date-time":"2025-12-09T06:20:22Z","timestamp":1765261222000},"source":"Crossref","is-referenced-by-count":1,"title":["Loss of cilia drives centriole clustering and elimination during mammalian spermatogenesis"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8735-3551","authenticated-orcid":false,"given":"Jun Jie","family":"Chen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangyu","family":"Gong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6881-8130","authenticated-orcid":false,"given":"Michael","family":"Mak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-6381-2201","authenticated-orcid":false,"given":"Feng-Qian","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9657-9966","authenticated-orcid":false,"given":"Ken-Ichi","family":"Takemaru","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025121103000855000_2025.12.04.692363v1.1","doi-asserted-by":"crossref","unstructured":"Asencio-Barria C , Defamie N , Saez JC , Mesnil M , Godoy AS (2019) Direct Intercellular Communications and Cancer: A Snapshot of the Biological Roles of Connexins in Prostate Cancer. Cancers (Basel ) 11","DOI":"10.3390\/cancers11091370"},{"key":"2025121103000855000_2025.12.04.692363v1.2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.74.1.68"},{"key":"2025121103000855000_2025.12.04.692363v1.3","doi-asserted-by":"crossref","unstructured":"Bonente D , Callaini G , Riparbelli MG (2025) Inactivation and Elimination of Centrioles During Development in the Genus Drosophila: Current Insights and Open Questions. Cells 14","DOI":"10.3390\/cells14120865"},{"key":"2025121103000855000_2025.12.04.692363v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2013.08.043"},{"key":"2025121103000855000_2025.12.04.692363v1.5","doi-asserted-by":"crossref","unstructured":"Buglak DB , Galletta BJ , Rusan NM (2025) Molecular models of the sperm head-tail coupling apparatus. J Cell Sci 138","DOI":"10.1242\/jcs.264168"},{"key":"2025121103000855000_2025.12.04.692363v1.6","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201406140"},{"key":"2025121103000855000_2025.12.04.692363v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.06.028"},{"key":"2025121103000855000_2025.12.04.692363v1.8","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.186221"},{"key":"2025121103000855000_2025.12.04.692363v1.9","doi-asserted-by":"crossref","unstructured":"Dia F , Strange T , Liang J , Hamilton J , Berkowitz KM (2017) Preparation of Meiotic Chromosome Spreads from Mouse Spermatocytes. J Vis Exp","DOI":"10.3791\/55378-v"},{"key":"2025121103000855000_2025.12.04.692363v1.10","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-09182-1"},{"key":"2025121103000855000_2025.12.04.692363v1.11","doi-asserted-by":"publisher","DOI":"10.1055\/s-2007-1019265"},{"key":"2025121103000855000_2025.12.04.692363v1.12","doi-asserted-by":"publisher","DOI":"10.1111\/andr.12165"},{"key":"2025121103000855000_2025.12.04.692363v1.13","doi-asserted-by":"crossref","first-page":"1001689","DOI":"10.3389\/fcell.2022.1001689","article-title":"Uncoupling cell division and cytokinesis during germline development in metazoans","volume":"10","year":"2022","journal-title":"Front Cell Dev Biol"},{"key":"2025121103000855000_2025.12.04.692363v1.14","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a005850"},{"key":"2025121103000855000_2025.12.04.692363v1.15","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00013.2015"},{"key":"2025121103000855000_2025.12.04.692363v1.16","doi-asserted-by":"crossref","unstructured":"Han C (2024) Gene expression programs in mammalian spermatogenesis. Development 151","DOI":"10.1242\/dev.202033"},{"key":"2025121103000855000_2025.12.04.692363v1.17","doi-asserted-by":"crossref","unstructured":"Hannaford MR , Rusan NM (2024) Positioning centrioles and centrosomes. J Cell Biol 223","DOI":"10.1083\/jcb.202311140"},{"key":"2025121103000855000_2025.12.04.692363v1.18","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2022.897029"},{"key":"2025121103000855000_2025.12.04.692363v1.19","doi-asserted-by":"crossref","unstructured":"Hilgendorf KI , Myers BR , Reiter JF (2024) Emerging mechanistic understanding of cilia function in cellular signalling. Nat Rev Mol Cell Biol","DOI":"10.1038\/s41580-023-00698-5"},{"key":"2025121103000855000_2025.12.04.692363v1.20","doi-asserted-by":"publisher","DOI":"10.1002\/dvg.22937"},{"key":"2025121103000855000_2025.12.04.692363v1.21","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1038\/s42003-021-02171-5","article-title":"WDR62 is required for centriole duplication in spermatogenesis and manchette removal in spermiogenesis","volume":"4","year":"2021","journal-title":"Commun Biol"},{"key":"2025121103000855000_2025.12.04.692363v1.22","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1530\/REP-21-0042","article-title":"CEP164 is essential for efferent duct multiciliogenesis and male fertility","volume":"162","year":"2021","journal-title":"Reproduction"},{"key":"2025121103000855000_2025.12.04.692363v1.23","doi-asserted-by":"crossref","unstructured":"Hoque M , Li FQ , Weber WD , Chen JJ , Kim EN , Kuo PL , Visconti PE , Takemaru KI (2024) The Cby3\/ciBAR1 complex positions the annulus along the sperm flagellum during spermiogenesis. J Cell Biol 223","DOI":"10.1083\/jcb.202307147"},{"key":"2025121103000855000_2025.12.04.692363v1.24","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1016\/j.cub.2022.12.046","article-title":"Drosophila transition fibers are essential for IFT-dependent ciliary elongation but not basal body docking and ciliary budding","volume":"33","year":"2023","journal-title":"Curr Biol"},{"key":"2025121103000855000_2025.12.04.692363v1.25","doi-asserted-by":"crossref","unstructured":"Hyland RM , Brody SL (2021) Impact of Motile Ciliopathies on Human Development and Clinical Consequences in the Newborn. Cells 11","DOI":"10.3390\/cells11010125"},{"key":"2025121103000855000_2025.12.04.692363v1.26","doi-asserted-by":"crossref","unstructured":"Kanie T , Liu B , Love JF , Fisher SD , Gustavsson AK , Jackson PK (2025) A hierarchical pathway for assembly of the distal appendages that organize primary cilia. Elife 14","DOI":"10.7554\/eLife.85999"},{"key":"2025121103000855000_2025.12.04.692363v1.27","doi-asserted-by":"publisher","DOI":"10.1038\/s44319-024-00260-0"},{"key":"2025121103000855000_2025.12.04.692363v1.28","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth1204-249"},{"key":"2025121103000855000_2025.12.04.692363v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.mce.2013.06.030"},{"key":"2025121103000855000_2025.12.04.692363v1.30","doi-asserted-by":"publisher","DOI":"10.1126\/science.aad2156"},{"key":"2025121103000855000_2025.12.04.692363v1.31","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.00160-16"},{"key":"2025121103000855000_2025.12.04.692363v1.32","doi-asserted-by":"crossref","first-page":"230228","DOI":"10.1098\/rsob.230228","article-title":"The role of cilia during organogenesis in zebrafish","volume":"13","year":"2023","journal-title":"Open Biol"},{"key":"2025121103000855000_2025.12.04.692363v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2019.04.002"},{"key":"2025121103000855000_2025.12.04.692363v1.34","doi-asserted-by":"crossref","unstructured":"Lopez-Jimenez P , Perez-Martin S , Hidalgo I , Garcia-Gonzalo FR , Page J , Gomez R (2022) The Male Mouse Meiotic Cilium Emanates from the Mother Centriole at Zygotene Prior to Centrosome Duplication. Cells 12","DOI":"10.1101\/2022.10.20.512932"},{"key":"2025121103000855000_2025.12.04.692363v1.35","doi-asserted-by":"crossref","first-page":"1114769","DOI":"10.3389\/fcell.2023.1114769","article-title":"What do we know about blood-testis barrier? current understanding of its structure and physiology","volume":"11","year":"2023","journal-title":"Front Cell Dev Biol"},{"key":"2025121103000855000_2025.12.04.692363v1.36","doi-asserted-by":"crossref","unstructured":"Ma D , Wang F , Teng J , Huang N , Chen J (2023) Structure and function of distal and subdistal appendages of the mother centriole. J Cell Sci 136","DOI":"10.1242\/jcs.260560"},{"key":"2025121103000855000_2025.12.04.692363v1.37","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms8676."},{"key":"2025121103000855000_2025.12.04.692363v1.38","doi-asserted-by":"publisher","DOI":"10.1038\/s41576-023-00587-9"},{"key":"2025121103000855000_2025.12.04.692363v1.39","doi-asserted-by":"publisher","DOI":"10.1007\/s10555-020-09937-z"},{"key":"2025121103000855000_2025.12.04.692363v1.40","doi-asserted-by":"publisher","DOI":"10.1038\/s41581-023-00773-2"},{"key":"2025121103000855000_2025.12.04.692363v1.41","doi-asserted-by":"publisher","DOI":"10.1210\/er.2014-1101"},{"key":"2025121103000855000_2025.12.04.692363v1.42","doi-asserted-by":"crossref","unstructured":"Mytlis A , Kumar V , Qiu T , Deis R , Hart N , Levy K , Masek M , Shawahny A , Ahmad A , Eitan H et al (2022) Control of meiotic chromosomal bouquet and germ cell morphogenesis by the zygotene cilium. Science 376: eabh3104","DOI":"10.1126\/science.abh3104"},{"key":"2025121103000855000_2025.12.04.692363v1.43","doi-asserted-by":"crossref","first-page":"102158","DOI":"10.1016\/j.ceb.2023.102158","article-title":"The many faces of the bouquet centrosome MTOC in meiosis and germ cell development","volume":"81","year":"2023","journal-title":"Curr Opin Cell Biol"},{"key":"2025121103000855000_2025.12.04.692363v1.44","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2022.05.001"},{"key":"2025121103000855000_2025.12.04.692363v1.45","doi-asserted-by":"publisher","DOI":"10.1016\/j.semcdb.2021.06.016"},{"key":"2025121103000855000_2025.12.04.692363v1.46","doi-asserted-by":"publisher","DOI":"10.1002\/aja.1000990307"},{"key":"2025121103000855000_2025.12.04.692363v1.47","doi-asserted-by":"publisher","DOI":"10.4161\/cc.8.4.7844"},{"key":"2025121103000855000_2025.12.04.692363v1.48","doi-asserted-by":"publisher","DOI":"10.1002\/dvg.20437"},{"key":"2025121103000855000_2025.12.04.692363v1.49","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E15-08-0578"},{"key":"2025121103000855000_2025.12.04.692363v1.50","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201202126"},{"key":"2025121103000855000_2025.12.04.692363v1.51","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2009.0206"},{"key":"2025121103000855000_2025.12.04.692363v1.52","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1007128"},{"key":"2025121103000855000_2025.12.04.692363v1.53","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1627708"},{"key":"2025121103000855000_2025.12.04.692363v1.54","doi-asserted-by":"publisher","DOI":"10.1038\/s44319-024-00187-6"},{"key":"2025121103000855000_2025.12.04.692363v1.55","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsbmb.2008.03.004"},{"key":"2025121103000855000_2025.12.04.692363v1.56","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00009.2020"},{"key":"2025121103000855000_2025.12.04.692363v1.57","doi-asserted-by":"publisher","DOI":"10.1111\/j.1582-4934.2004.tb00328.x"},{"key":"2025121103000855000_2025.12.04.692363v1.58","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1038\/s41572-020-0209-6","article-title":"Motile ciliopathies","volume":"6","year":"2020","journal-title":"Nat Rev Dis Primers"},{"key":"2025121103000855000_2025.12.04.692363v1.59","doi-asserted-by":"crossref","unstructured":"Wellard SR , Hopkins J , Jordan PW (2018) A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model. J Vis Exp","DOI":"10.3791\/56453-v"},{"key":"2025121103000855000_2025.12.04.692363v1.60","doi-asserted-by":"publisher","DOI":"10.15252\/embr.202051023"},{"key":"2025121103000855000_2025.12.04.692363v1.61","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1186\/s12610-024-00241-2","article-title":"The annulus: composition, role and importance in sperm flagellum biogenesis and male fertility","volume":"34","year":"2024","journal-title":"Basic Clin Androl"},{"key":"2025121103000855000_2025.12.04.692363v1.62","doi-asserted-by":"publisher","DOI":"10.1093\/biolre\/ioaa016"},{"key":"2025121103000855000_2025.12.04.692363v1.63","doi-asserted-by":"crossref","unstructured":"Xie H , Wang X , Jin M , Li L , Zhu J , Kang Y , Chen Z , Sun Y , Zhao C (2022) Cilia regulate meiotic recombination in zebrafish. J Mol Cell Biol 14","DOI":"10.1093\/jmcb\/mjac049"},{"key":"2025121103000855000_2025.12.04.692363v1.64","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E16-05-0318"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2025.12.04.692363","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T11:00:39Z","timestamp":1765450839000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2025.12.04.692363"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,8]]},"references-count":64,"URL":"https:\/\/doi.org\/10.64898\/2025.12.04.692363","relation":{},"subject":[],"published":{"date-parts":[[2025,12,8]]},"subtype":"preprint"}}