{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,8]],"date-time":"2026-02-08T09:20:17Z","timestamp":1770542417225,"version":"3.49.0"},"reference-count":35,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2024,11,19]],"date-time":"2024-11-19T00:00:00Z","timestamp":1731974400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["R01-HD105944"],"award-info":[{"award-number":["R01-HD105944"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Cancer Institute of the National Institutes of Health","award":["P30CA046592"],"award-info":[{"award-number":["P30CA046592"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82101674"],"award-info":[{"award-number":["82101674"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Human Development of the National Institutes of Health","award":["RO1 HD114311 (Z.Z.)"],"award-info":[{"award-number":["RO1 HD114311 (Z.Z.)"]}]},{"name":"Eunice Kennedy Shriver National Institute of Child Health","award":["R01-HD105944"],"award-info":[{"award-number":["R01-HD105944"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,2,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Intraflagellar transport 25 is a component of the intraflagellar transport 25-B complex. In mice, even though this intraflagellar transport component is not required for cilia formation in somatic cells, it is essential for sperm formation. However, the intracellular localization of this protein in male germ cells is not known given no reliable antibodies are available for histologic studies, and the dynamic trafficking in the developing sperm flagella is not clear. To examine localization of the protein in male germ cells and further investigate the mechanism of intraflagellar transport in sperm formation, particularly to look into the dynamic trafficking of the protein, we generated a mouse intraflagellar transport 25\u2013green fluorescent protein knock-in mouse model using the clustered regularly interspaced short palindromic repeats\/clustered regularly interspaced short palindromic repeats associated protein 9 system, with the mouse intraflagellar transport 25 protein fused with a green fluorescent protein tag in the C-terminus. Three independent lines were analyzed. Western blotting using both anti-intraflagellar transport 25 and anti-green fluorescent protein antibodies showed that the intraflagellar transport 25\u2013green fluorescent protein fusion protein was highly abundant only in the testis, which is consistent with the endogenous intraflagellar transport 25 protein. Examination of localization of the intraflagellar transport 25\u2013green fluorescent protein in isolated germ cells revealed that the fusion protein was present in the cytoplasm of spermatocytes and round spermatids and a strong signal was present in the developing sperm flagellar. The homozygous knock-in mice had normal spermatogenesis, fertility and sperm parameters. Diffusion analysis of intraflagellar transport 25 within the developing flagellar revealed the presence of both mobile and immobile fractions as revealed by fluorescence recovery after photobleaching. Kymograph and fluorescence recovery after photobleaching analyses demonstrate the transport of intraflagellar transport 25\u2013green fluorescent protein within the developing tail demonstrate no apparent preference for trafficking toward and away from the cell body. The speed of trafficking depends on the stage of sperm development, ranging from highly mobile unrestricted diffusion initially, mobile punctate structures in developing sperm, and immobile punctate structures in mature sperm. Our studies demonstrate that mouse intraflagellar transport 25 travels along the developing sperm flagella in two directions that might be essential for functional sperm formation.<\/jats:p>","DOI":"10.1093\/biolre\/ioae171","type":"journal-article","created":{"date-parts":[[2024,11,19]],"date-time":"2024-11-19T18:15:37Z","timestamp":1732040137000},"page":"309-318","source":"Crossref","is-referenced-by-count":1,"title":["Two-directional trafficking of the IFT25 protein in the developing mouse sperm flagella"],"prefix":"10.1093","volume":"112","author":[{"given":"Wei","family":"Li","sequence":"first","affiliation":[{"name":"Department of Physiology, Wayne State University , Detroit, MI ,","place":["USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Changmin","family":"Niu","sequence":"additional","affiliation":[{"name":"Department of Physiology, Wayne State University , Detroit, MI ,","place":["USA"]},{"name":"School of Nursing School of Public Health, Yangzhou University , Yangzhou, Jiangsu ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yi Tian","family":"Yap","sequence":"additional","affiliation":[{"name":"Department of Physiology, Wayne State University , Detroit, MI ,","place":["USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tao","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Physiology, Wayne State University , Detroit, MI ,","place":["USA"]},{"name":"Institute of Reproductive Health , Tongji Medical College, , Wuhan ,","place":["PR China"]},{"name":"Huazhong University of Science and Technology , Tongji Medical College, , Wuhan ,","place":["PR China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cheng","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Physiology, Wayne State University , Detroit, MI ,","place":["USA"]},{"name":"Department of Occupational and Environmental Health , School of Public Health, , Wuhan, Hubei ,","place":["PR China"]},{"name":"Wuhan University of Science and Technology , School of Public Health, , Wuhan, Hubei ,","place":["PR China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mariska","family":"Goswami","sequence":"additional","affiliation":[{"name":"Troy High School , Troy, Michigan ,","place":["USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sanjana","family":"Kandiraju","sequence":"additional","affiliation":[{"name":"International Academy High School , Oakland County, Michigan ,","place":["USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Opeyemi","family":"Dhikhirullahi","sequence":"additional","affiliation":[{"name":"Department of Physiology, Wayne State University , Detroit, MI ,","place":["USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jie","family":"Xu","sequence":"additional","affiliation":[{"name":"Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical School , Ann Arbor, MI ,","place":["USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jifeng","family":"Zhang","sequence":"additional","affiliation":[{"name":"Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical School , Ann Arbor, MI ,","place":["USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher V","family":"Kelly","sequence":"additional","affiliation":[{"name":"Department of Physics and Astronomy, Wayne State University , Detroit, MI ,","place":["USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhibing","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Physiology, Wayne State University , Detroit, MI ,","place":["USA"]},{"name":"Department of Obstetrics and Gynecology, Wayne State University , Detroit, MI ,","place":["USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2024,11,19]]},"reference":[{"key":"2025042607515186600_ref1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13630-017-0046-8","article-title":"CILIA: before and after","volume":"6","author":"Satir","year":"2017","journal-title":"Cilia"},{"key":"2025042607515186600_ref2","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1038\/s41576-023-00587-9","article-title":"Primary cilia as dynamic and diverse signalling hubs in development and disease","volume":"24","author":"Mill","year":"2023","journal-title":"Nat Rev Genet"},{"key":"2025042607515186600_ref3","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1038\/s41572-020-0209-6","article-title":"Motile ciliopathies","volume":"6","author":"Wallmeier","year":"2020","journal-title":"Nat Rev Dis Primers"},{"key":"2025042607515186600_ref4","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/S0070-2153(08)00802-8","article-title":"Intraflagellar transport (IFT) role in ciliary assembly, resorption and signaling","volume":"85","author":"Pedersen","year":"2008","journal-title":"Curr Top Dev Biol"},{"key":"2025042607515186600_ref5","doi-asserted-by":"crossref","first-page":"a021998","DOI":"10.1101\/cshperspect.a021998","article-title":"Intraflagellar transport and ciliary dynamics","volume":"9","author":"Ishikawa","year":"2017","journal-title":"Cold Spring Harb Perspect Biol"},{"key":"2025042607515186600_ref6","doi-asserted-by":"crossref","first-page":"a028092","DOI":"10.1101\/cshperspect.a028092","article-title":"The intraflagellar transport machinery","volume":"8","author":"Taschner","year":"2016","journal-title":"Cold Spring Harb Perspect Biol"},{"key":"2025042607515186600_ref7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1002\/cm.10035","article-title":"A healthy understanding of intraflagellar transport","volume":"52","author":"Sloboda","year":"2002","journal-title":"Cell Motil Cytoskeleton"},{"key":"2025042607515186600_ref8","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1016\/S0962-8924(02)02410-8","article-title":"Intraflagellar transport and cilia-dependent diseases","volume":"12","author":"Pazour","year":"2002","journal-title":"Trends Cell Biol"},{"key":"2025042607515186600_ref9","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1146\/annurev-cellbio-120219-034238","article-title":"Structural biology of cilia and intraflagellar transport","volume":"38","author":"Klena","year":"2022","journal-title":"Annu Rev Cell Dev Biol"},{"key":"2025042607515186600_ref10","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/j.ceb.2018.03.004","article-title":"The molecular machines that traffic signaling receptors into and out of cilia","volume":"51","author":"Nachury","year":"2018","journal-title":"Curr Opin Cell Biol"},{"key":"2025042607515186600_ref11","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/j.devcel.2014.09.011","article-title":"IFT27 links the BBSome to IFT for maintenance of the ciliary signaling compartment","volume":"31","author":"Eguether","year":"2014","journal-title":"Dev Cell"},{"key":"2025042607515186600_ref12","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1016\/j.devcel.2012.04.009","article-title":"IFT25 links the signal-dependent movement of hedgehog components to intraflagellar transport","volume":"22","author":"Keady","year":"2012","journal-title":"Dev Cell"},{"key":"2025042607515186600_ref13","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1093\/biolre\/iox029","article-title":"IFT25, an intraflagellar transporter protein dispensable for ciliogenesis in somatic cells, is essential for sperm flagella formation","volume":"96","author":"Liu","year":"2017","journal-title":"Biol Reprod"},{"key":"2025042607515186600_ref14","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1093\/biolre\/iox096","article-title":"Formation and function of sperm tail structures in association with sperm motility defects","volume":"97","author":"Lehti","year":"2017","journal-title":"Biol Reprod"},{"key":"2025042607515186600_ref15","doi-asserted-by":"crossref","first-page":"12946","DOI":"10.1073\/pnas.0404280101","article-title":"Haploinsufficiency for the murine orthologue of Chlamydomonas PF20 disrupts spermatogenesis","volume":"101","author":"Zhang","year":"2004","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2025042607515186600_ref16","doi-asserted-by":"crossref","first-page":"17055","DOI":"10.1073\/pnas.0906414106","article-title":"MEIG1 is essential for spermiogenesis in mice","volume":"106","author":"Zhang","year":"2009","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2025042607515186600_ref17","doi-asserted-by":"crossref","first-page":"e11","DOI":"10.14440\/jbm.2014.36","article-title":"Advanced methods of microscope control using \u03bcManager software","volume":"1","author":"Edelstein","year":"2014","journal-title":"J Biol Methods"},{"key":"2025042607515186600_ref18","doi-asserted-by":"crossref","first-page":"1055","DOI":"10.1016\/S0006-3495(76)85755-4","article-title":"Mobility measurement by analysis of fluorescence photobleaching recovery kinetics","volume":"16","author":"Axelrod","year":"1976","journal-title":"Biophys J"},{"key":"2025042607515186600_ref19","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1901\/jeab.2004.82-329","article-title":"Steps and pips in the history of the cumulative recorder","volume":"82","author":"Lattal","year":"2004","journal-title":"J Exp Anal Behav"},{"key":"2025042607515186600_ref20","doi-asserted-by":"crossref","first-page":"6943","DOI":"10.1073\/pnas.1221011110","article-title":"Evolution of modular intraflagellar transport from a coatomer-like progenitor","volume":"110","author":"Dam","year":"2013","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2025042607515186600_ref21","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1002\/cm.20369","article-title":"HA-tagging of putative flagellar proteins in Chlamydomonas reinhardtii identifies a novel protein of intraflagellar transport complex B","volume":"66","author":"Lechtreck","year":"2009","journal-title":"Cell Motil Cytoskeleton"},{"key":"2025042607515186600_ref22","doi-asserted-by":"crossref","first-page":"1130","DOI":"10.1016\/j.kint.2017.04.027","article-title":"A novel model of autosomal recessive polycystic kidney questions the role of the fibrocystin C-terminus in disease mechanism","volume":"92","author":"Outeda","year":"2017","journal-title":"Kidney Int"},{"key":"2025042607515186600_ref23","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/2046-2530-2-8","article-title":"An inducible CiliaGFP mouse model for in vivo visualization and analysis of cilia in live tissue","volume":"2","author":"O\u2019Connor","year":"2013","journal-title":"Cilia"},{"key":"2025042607515186600_ref24","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1111\/tpj.16265","article-title":"Efficient methods for multiple types of precise gene-editing in Chlamydomonas","volume":"115","author":"Chen","year":"2023","journal-title":"Plant J"},{"key":"2025042607515186600_ref25","doi-asserted-by":"crossref","first-page":"1725","DOI":"10.1242\/dev.00407","article-title":"The left- right determinant inversin is a component of node monocilia and other 9+0 cilia","volume":"130","author":"Watanabe","year":"2003","journal-title":"Development"},{"key":"2025042607515186600_ref26","doi-asserted-by":"crossref","first-page":"18274","DOI":"10.1038\/s41598-021-97046-4","article-title":"Cystin genetic variants cause autosomal recessive polycystic kidney disease associated with altered Myc expression","volume":"11","author":"Yang","year":"2021","journal-title":"Sci Rep"},{"key":"2025042607515186600_ref27","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1038\/ncb3091","article-title":"Lineage specificity of primary cilia in the mouse embryo","volume":"17","author":"Bangs","year":"2015","journal-title":"Nat Cell Biol"},{"key":"2025042607515186600_ref28","doi-asserted-by":"crossref","first-page":"1623","DOI":"10.3390\/cells10071623","article-title":"Identification of cilia in different mouse tissues","volume":"10","author":"Li","year":"2021","journal-title":"Cells"},{"key":"2025042607515186600_ref29","doi-asserted-by":"crossref","first-page":"e23355","DOI":"10.1002\/dvg.23355","article-title":"A rat H1t-GFP transgene recapitulates endogenous H1t expression pattern in mouse","volume":"58","author":"Lipschutz","year":"2020","journal-title":"Genesis"},{"key":"2025042607515186600_ref30","first-page":"1","article-title":"Introduction of genes into the germ line of animals","volume":"80","author":"Brinster","year":"1984","journal-title":"Harvey Lect"},{"key":"2025042607515186600_ref31","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1146\/annurev.ge.20.120186.002341","article-title":"Germ-line transformation of mice","volume":"20","author":"Palmiter","year":"1986","journal-title":"Annu Rev Genet"},{"key":"2025042607515186600_ref32","doi-asserted-by":"crossref","first-page":"jcs228296","DOI":"10.1242\/jcs.228296","article-title":"IFT25 is required for the construction of the trypanosome flagellum","volume":"132","author":"Huet","year":"2019","journal-title":"J Cell Sci"},{"key":"2025042607515186600_ref33","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1002\/bies.200900031","article-title":"The motile cilium in development and disease: emerging new insights","volume":"31","author":"Roy","year":"2009","journal-title":"Bioessays"},{"key":"2025042607515186600_ref34","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1002\/path.4843","article-title":"Motile and non-motile cilia in human pathology: from function to phenotypes","volume":"241","author":"Mitchison","year":"2017","journal-title":"J Pathol"},{"key":"2025042607515186600_ref35","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1002\/jemt.20787","article-title":"Surfing the wave, cycle, life history, and genes\/proteins expressed by testicular germ cells. Part 2: changes in spermatid organelles associated with development of spermatozoa","volume":"73","author":"Hermo","year":"2010","journal-title":"Microsc Res Tech"}],"container-title":["Biology of Reproduction"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/biolreprod\/advance-article-pdf\/doi\/10.1093\/biolre\/ioae171\/60957998\/ioae171.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/biolreprod\/article-pdf\/112\/2\/309\/60957998\/ioae171.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/biolreprod\/article-pdf\/112\/2\/309\/60957998\/ioae171.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,26]],"date-time":"2025-04-26T11:52:02Z","timestamp":1745668322000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/biolreprod\/article\/112\/2\/309\/7904804"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,19]]},"references-count":35,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2024,11,19]]},"published-print":{"date-parts":[[2025,2,14]]}},"URL":"https:\/\/doi.org\/10.1093\/biolre\/ioae171","relation":{},"ISSN":["0006-3363","1529-7268"],"issn-type":[{"value":"0006-3363","type":"print"},{"value":"1529-7268","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2025,2]]},"published":{"date-parts":[[2024,11,19]]}}}