{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T20:41:06Z","timestamp":1760560866170,"version":"build-2065373602"},"reference-count":60,"publisher":"Oxford University Press (OUP)","issue":"4","funder":[{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["R01 HD10808"],"award-info":[{"award-number":["R01 HD10808"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,10,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Sertoli cells are essential to successful, continuous sperm production and are responsible for many processes throughout spermatogenesis, including germ cell movement and compartmentalization of the seminiferous tubule interior. These functions due to the F-actin cytoskeleton present within the seminiferous tubules, which facilitates cell movement and adhesion. While some molecular players that regulate the testicular F-actin cytoskeleton are known, the expression of other actin-related genes in the mammalian testis remains unknown. In this study, we revisited previously published next-generation sequencing data and combined this with immunohistochemical analysis to identify the cell- and stage-specific expression of multiple alpha-actinin and myosin genes and their protein products. This work provides the first characterization of alpha-actinin and myosin localization within murine testes throughout spermatogenesis. We identified alpha-actinin-3, alpha-actinin-4, myosin VIIa (MYO7A), and myosin 10 as potential functional candidates regulating cytoskeleton dynamics throughout spermatogenesis based on the high mRNA expression of these genes observed within Sertoli cells, as well as the specific protein localization observed within Sertoli cells. We additionally found expression of MYO7A within early spermatogonia and meiotic germ cells, suggesting a potential role of these proteins during mitotic and meiotic division. Overall, this study provides insight into the expression of many F-actin-related genes during mammalian testicular maturation and contributes to our understanding of dynamic Sertoli cell gene expression in mammalian testes.<\/jats:p>","DOI":"10.1093\/biolre\/ioaf086","type":"journal-article","created":{"date-parts":[[2025,4,9]],"date-time":"2025-04-09T07:31:08Z","timestamp":1744183868000},"page":"934-943","source":"Crossref","is-referenced-by-count":0,"title":["Testicular stage- and cell-specific expression of F-actin-binding proteins"],"prefix":"10.1093","volume":"113","author":[{"given":"Shelby L","family":"Havel","sequence":"first","affiliation":[{"name":"School of Molecular Biosciences, Washington State University , Pullman, Washington ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert J","family":"Hubbard","sequence":"additional","affiliation":[{"name":"School of Molecular Biosciences, Washington State University , Pullman, Washington ,","place":["United States"]},{"name":"Department of Chemistry, University of Idaho , Moscow, Idaho ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael D","family":"Griswold","sequence":"additional","affiliation":[{"name":"School of Molecular Biosciences, Washington State University , Pullman, Washington ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2025,4,10]]},"reference":[{"key":"2025101516134689200_ref1","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1111\/j.1749-6632.1952.tb26576.x","article-title":"Definition of the stages of the cycle of the seminiferous epithelium in the rat","volume":"55","author":"Leblond","year":"1952","journal-title":"Ann NY Acad Sci"},{"key":"2025101516134689200_ref2","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1095\/biolreprod43.3.517","article-title":"Frequency of the stages in the cycle of the seminiferous epithelium in the rat","volume":"43","author":"Hess","year":"1990","journal-title":"Biol Reprod"},{"key":"2025101516134689200_ref3","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1111\/j.1365-2605.1993.tb01156.x","article-title":"Histological and histopathological evaluation of the testis","volume":"16","author":"Russell","year":"1993","journal-title":"Int J Androl"},{"key":"2025101516134689200_ref4","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1095\/biolreprod.116.141770","article-title":"Characterizing the spermatogonial response to retinoic acid during the onset of spermatogenesis and following synchronization in the neonatal mouse testis","volume":"95","author":"Agrimson","year":"2016","journal-title":"Biol Reprod"},{"key":"2025101516134689200_ref5","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1152\/physrev.00013.2015","article-title":"Spermatogenesis: the commitment to meiosis","volume":"96","author":"Griswold","year":"2016","journal-title":"Physiol Rev"},{"key":"2025101516134689200_ref6","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1111\/andr.12165","article-title":"The Sertoli cell: one hundred fifty years of beauty and plasticity","volume":"4","author":"Fran\u00e7a","year":"2016","journal-title":"Andrology"},{"key":"2025101516134689200_ref7","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1210\/er.2014-1101","article-title":"The mammalian blood\u2013testis barrier: its biology and regulation","volume":"36","author":"Mruk","year":"2015","journal-title":"Endocr Rev"},{"key":"2025101516134689200_ref8","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1210\/er.2003-0022","article-title":"Sertoli\u2013Sertoli and Sertoli\u2013germ cell interactions and their significance in germ cell movement in the seminiferous epithelium during spermatogenesis","volume":"25","author":"Mruk","year":"2004","journal-title":"Endocr Rev"},{"key":"2025101516134689200_ref9","doi-asserted-by":"crossref","first-page":"798","DOI":"10.1126\/science.1219969","article-title":"Germ cell migration across Sertoli cell tight junctions","volume":"338","author":"Smith","year":"2012","journal-title":"Science"},{"key":"2025101516134689200_ref10","first-page":"186","volume-title":"Molecular Mechanisms in Spermatogenesis","author":"Vogl","year":"2008"},{"key":"2025101516134689200_ref11","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.semcdb.2016.01.004","article-title":"Regulation of microtubule (MT)-based cytoskeleton in the seminiferous epithelium during spermatogenesis","volume":"59","author":"Tang","year":"2016","journal-title":"Semin Cell Dev Biol"},{"key":"2025101516134689200_ref12","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.reprotox.2020.05.017","article-title":"Actin binding proteins, actin cytoskeleton and spermatogenesis\u2014lesson from toxicant models","volume":"96","author":"Wang","year":"2020","journal-title":"Reprod Toxicol"},{"key":"2025101516134689200_ref13","first-page":"381","volume-title":"Advances in Protein Chemistry and Structural Biology","author":"Wang","year":"2024"},{"key":"2025101516134689200_ref14","doi-asserted-by":"crossref","first-page":"1907","DOI":"10.1210\/en.2012-2269","article-title":"Palladin is a regulator of actin filament bundles at the ectoplasmic specialization in adult rat testes","volume":"154","author":"Qian","year":"2013","journal-title":"Endocrinology"},{"key":"2025101516134689200_ref15","doi-asserted-by":"crossref","first-page":"11411","DOI":"10.1073\/pnas.1001823107","article-title":"Restricted Arp3 expression in the testis prevents blood\u2013testis barrier disruption during junction restructuring at spermatogenesis","volume":"107","author":"Lie","year":"2010","journal-title":"Proc Natl Acad Sci"},{"key":"2025101516134689200_ref16","doi-asserted-by":"crossref","first-page":"937","DOI":"10.1111\/andr.12076","article-title":"Actin-related protein 2\/3 complex-based actin polymerization is critical for male fertility","volume":"3","author":"Lee","year":"2015","journal-title":"Andrology"},{"key":"2025101516134689200_ref17","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/j.autrev.2010.12.009","article-title":"Alpha-actinin: a multidisciplinary protein with important role in B-cell driven autoimmunity","volume":"10","author":"Oikonomou","year":"2011","journal-title":"Autoimmun Rev"},{"key":"2025101516134689200_ref18","volume-title":"The Cell: A Molecular Approach","author":"Cooper","year":"2000","edition":"2nd"},{"key":"2025101516134689200_ref19","doi-asserted-by":"crossref","first-page":"2688","DOI":"10.1007\/s00018-008-8080-8","article-title":"\u03b1-Actinin structure and regulation","volume":"65","author":"Sj\u00f6blom","year":"2008","journal-title":"Cell Mol Life Sci"},{"key":"2025101516134689200_ref20","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1186\/s13578-015-0029-7","article-title":"The actinin family of actin cross-linking proteins\u2014a genetic perspective","volume":"5","author":"Murphy","year":"2015","journal-title":"Cell Biosci"},{"key":"2025101516134689200_ref21","doi-asserted-by":"crossref","first-page":"1591","DOI":"10.1093\/biolre\/ioab160","article-title":"Two distinct Sertoli cell states are regulated via germ cell crosstalk","volume":"105","author":"Gewiss","year":"2021","journal-title":"Biol Reprod"},{"key":"2025101516134689200_ref22","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1002\/ar.1092330109","article-title":"Localization of actin, alpha-actinin, and tropomyosin in bovine spermatozoa and epididymal epithelium","volume":"233","author":"Yagi","year":"1992","journal-title":"Anat Rec"},{"key":"2025101516134689200_ref23","doi-asserted-by":"crossref","first-page":"9730","DOI":"10.1038\/s41598-019-46324-3","article-title":"Testes and duct deferens of mice during space flight: cytoskeleton structure, sperm-specific proteins and epigenetic events","volume":"9","author":"Ogneva","year":"2019","journal-title":"Sci Rep"},{"key":"2025101516134689200_ref24","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1111\/azo.12422","article-title":"Histological study and immunohistochemical location of cytoskeletal proteins in the testis and epididymis of the three species of lizards of the family Leiosauridae (Reptilia: Squamata)","volume":"104","author":"Firmiano","year":"2022","journal-title":"Acta Zool"},{"key":"2025101516134689200_ref25","volume-title":"The Cell: A Molecular Approach","author":"Cooper","year":"2000","edition":"2nd"},{"key":"2025101516134689200_ref26","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1210\/en.2018-00855","article-title":"Myosin VIIa supports spermatid\/organelle transport and cell adhesion during spermatogenesis in the rat testis","volume":"160","author":"Wen","year":"2019","journal-title":"Endocrinology"},{"key":"2025101516134689200_ref27","doi-asserted-by":"crossref","first-page":"D991","DOI":"10.1093\/nar\/gks1193","article-title":"NCBI GEO: archive for functional genomics data sets\u2014update","volume":"41","author":"Barrett","year":"2013","journal-title":"Nucleic Acids Res"},{"key":"2025101516134689200_ref28","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol"},{"key":"2025101516134689200_ref29","doi-asserted-by":"crossref","first-page":"dev183251","DOI":"10.1242\/dev.183251","article-title":"Single-cell RNAseq analysis of testicular germ and somatic cell development during the perinatal period","volume":"147","author":"Tan","year":"2020","journal-title":"Development"},{"key":"2025101516134689200_ref30","doi-asserted-by":"crossref","first-page":"dev202740","DOI":"10.1242\/dev.202740","article-title":"Destabilization of mRNAs enhances competence to initiate meiosis in mouse spermatogenic cells","volume":"151","author":"Pfaltzgraff","year":"2024","journal-title":"Development"},{"key":"2025101516134689200_ref31","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1016\/j.gpb.2021.08.001","article-title":"The genome sequence archive family: toward explosive data growth and diverse data types","volume":"19","author":"Chen","year":"2021","journal-title":"Genomics Proteomics Bioinformatics"},{"key":"2025101516134689200_ref32","doi-asserted-by":"crossref","first-page":"D18","DOI":"10.1093\/nar\/gkad1078","article-title":"Database resources of the National Genomics Data Center, China National Center for Bioinformation in 2024","volume":"52","author":"CNCB-NGDC Members and Partners","year":"2024","journal-title":"Nucleic Acids Res"},{"key":"2025101516134689200_ref33","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.jare.2022.12.007","article-title":"A single-cell transcriptomic landscape of mouse testicular aging","volume":"53","author":"Zhang","year":"2023","journal-title":"J Adv Res"},{"key":"2025101516134689200_ref34","doi-asserted-by":"crossref","first-page":"3573","DOI":"10.1016\/j.cell.2021.04.048","article-title":"Integrated analysis of multimodal single-cell data","volume":"184","author":"Hao","year":"2021","journal-title":"Cell"},{"key":"2025101516134689200_ref35","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1093\/biolre\/ioae115","article-title":"Temporal maturation of Sertoli cells during the establishment of the cycle of the seminiferous epithelium","volume":"111","author":"Havel","year":"2024","journal-title":"Biol Reprod"},{"key":"2025101516134689200_ref36","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1038\/s41422-018-0074-y","article-title":"Single-cell RNA-seq uncovers dynamic processes and critical regulators in mouse spermatogenesis","volume":"28","author":"Chen","year":"2018","journal-title":"Cell Res"},{"key":"2025101516134689200_ref37","doi-asserted-by":"crossref","first-page":"1650","DOI":"10.1016\/j.celrep.2018.10.026","article-title":"The mammalian spermatogenesis single-cell transcriptome, from spermatogonial stem cells to spermatids","volume":"25","author":"Hermann","year":"2018","journal-title":"Cell Rep"},{"key":"2025101516134689200_ref38","doi-asserted-by":"crossref","first-page":"1284184","DOI":"10.3389\/fcell.2023.1284184","article-title":"Core binding factor subunit \u03b2 plays diverse and essential roles in the male germline","volume":"11","author":"Rahmawati","year":"2023","journal-title":"Front Cell Dev Biol"},{"key":"2025101516134689200_ref39","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1016\/j.devcel.2018.07.025","article-title":"A comprehensive roadmap of murine spermatogenesis defined by single-cell RNA-seq","volume":"46","author":"Green","year":"2018","journal-title":"Dev Cell"},{"key":"2025101516134689200_ref40","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1016\/j.devcel.2020.05.010","article-title":"Single-cell RNA sequencing of human, macaque, and mouse testes uncovers conserved and divergent features of mammalian spermatogenesis","volume":"54","author":"Shami","year":"2020","journal-title":"Dev Cell"},{"key":"2025101516134689200_ref41","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1007\/978-1-62703-038-0_27","volume-title":"Spermatogenesis: Methods and Protocols","author":"Meistrich","year":"2013"},{"key":"2025101516134689200_ref42","doi-asserted-by":"crossref","DOI":"10.3389\/fgene.2019.00317","article-title":"Single-cell RNA-Seq technologies and related computational data analysis","volume":"10","author":"Chen","year":"2019","journal-title":"Front Genet"},{"key":"2025101516134689200_ref43","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1083\/jcb.200804062","article-title":"Myosin-10 and actin filaments are essential for mitotic spindle function","volume":"182","author":"Woolner","year":"2008","journal-title":"J Cell Biol"},{"key":"2025101516134689200_ref44","doi-asserted-by":"crossref","first-page":"10253","DOI":"10.1038\/ncomms10253","article-title":"F-actin mechanics control spindle centring in the mouse zygote","volume":"7","author":"Chaigne","year":"2016","journal-title":"Nat Commun"},{"key":"2025101516134689200_ref45","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1124\/pr.110.002790","article-title":"The blood\u2013testis barrier and its implications for male contraception","volume":"64","author":"Cheng","year":"2012","journal-title":"Pharmacol Rev"},{"key":"2025101516134689200_ref46","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1242\/jcs.s3-106.73.93","article-title":"Morphogenesis of the residual body of the mouse testis","volume":"s3-106","author":"Firlit","year":"1965","journal-title":"J Cell Sci"},{"key":"2025101516134689200_ref47","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.biochi.2024.11.004","article-title":"The completing of the second meiotic division by MII mouse oocytes correlates with the positioning of F-actin and mitochondria in the ooplasm","volume":"230","author":"Ryabukha","year":"2024","journal-title":"Biochimie"},{"key":"2025101516134689200_ref48","doi-asserted-by":"crossref","first-page":"2509","DOI":"10.1091\/mbc.9.9.2509","article-title":"Differential expression and functions of cortical myosin IIA and IIB isotypes during meiotic maturation, fertilization, and mitosis in mouse oocytes and embryos","volume":"9","author":"Simerly","year":"1998","journal-title":"MBoC"},{"key":"2025101516134689200_ref49","doi-asserted-by":"crossref","first-page":"1489","DOI":"10.1016\/S0960-9822(00)00828-9","article-title":"Myosin VI is required for asymmetric segregation of cellular components during C. elegans spermatogenesis","volume":"10","author":"Kelleher","year":"2000","journal-title":"Curr Biol"},{"key":"2025101516134689200_ref50","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1016\/S0960-9822(00)00420-6","article-title":"Fission yeast myosin-II isoforms assemble into contractile rings at distinct times during mitosis","volume":"10","author":"Bezanilla","year":"2000","journal-title":"Curr Biol"},{"key":"2025101516134689200_ref51","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/S0092-8674(04)00341-1","article-title":"Myosin II-dependent cortical movement is required for centrosome separation and positioning during mitotic spindle assembly","volume":"117","author":"Rosenblatt","year":"2004","journal-title":"Cell"},{"key":"2025101516134689200_ref52","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1083\/jcb.200709099","article-title":"The class V myosin motor protein, Myo2, plays a major role in mitochondrial motility in Saccharomyces cerevisiae","volume":"181","author":"Altmann","year":"2008","journal-title":"J Cell Biol"},{"key":"2025101516134689200_ref53","doi-asserted-by":"crossref","first-page":"R912","DOI":"10.1016\/j.cub.2008.08.043","article-title":"Mitosis: new roles for myosin-X and actin at the spindle","volume":"18","author":"W\u00fchr","year":"2008","journal-title":"Curr Biol"},{"key":"2025101516134689200_ref54","doi-asserted-by":"crossref","first-page":"3952","DOI":"10.1091\/mbc.e10-04-0293","article-title":"Ablation of nonmuscle myosin II-B and II-C reveals a role for nonmuscle myosin II in cardiac myocyte karyokinesis","volume":"21","author":"Ma","year":"2010","journal-title":"MBoC"},{"key":"2025101516134689200_ref55","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1016\/j.ydbio.2012.07.011","article-title":"Non-muscle myosin IIB is essential for cytokinesis during male meiotic cell divisions","volume":"369","author":"Yang","year":"2012","journal-title":"Dev Biol"},{"key":"2025101516134689200_ref56","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/j.gene.2015.10.022","article-title":"Myosin superfamily: the multi-functional and irreplaceable factors in spermatogenesis and testicular tumors","volume":"576","author":"Li","year":"2016","journal-title":"Gene"},{"key":"2025101516134689200_ref57","first-page":"1512","article-title":"Global signatures of protein and mRNA expression levels","volume":"5","author":"Sousa Abreu","year":"2009","journal-title":"Mol Biosyst"},{"key":"2025101516134689200_ref58","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1038\/nrg3185","article-title":"Insights into the regulation of protein abundance from proteomic and transcriptomic analyses","volume":"13","author":"Vogel","year":"2012","journal-title":"Nat Rev Genet"},{"key":"2025101516134689200_ref59","doi-asserted-by":"crossref","first-page":"3966","DOI":"10.1016\/j.febslet.2009.10.036","article-title":"Correlation of mRNA and protein in complex biological samples","volume":"583","author":"Maier","year":"2009","journal-title":"FEBS Lett"},{"key":"2025101516134689200_ref60","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1016\/j.cell.2018.01.035","article-title":"Pervasive, coordinated protein-level changes driven by transcript isoform switching during meiosis","volume":"172","author":"Cheng","year":"2018","journal-title":"Cell"}],"container-title":["Biology of Reproduction"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/biolreprod\/advance-article-pdf\/doi\/10.1093\/biolre\/ioaf086\/62903549\/ioaf086.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/biolreprod\/article-pdf\/113\/4\/934\/62903549\/ioaf086.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/biolreprod\/article-pdf\/113\/4\/934\/62903549\/ioaf086.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T20:14:01Z","timestamp":1760559241000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/biolreprod\/article\/113\/4\/934\/8110031"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,10]]},"references-count":60,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2025,4,10]]},"published-print":{"date-parts":[[2025,10,14]]}},"URL":"https:\/\/doi.org\/10.1093\/biolre\/ioaf086","relation":{},"ISSN":["0006-3363","1529-7268"],"issn-type":[{"type":"print","value":"0006-3363"},{"type":"electronic","value":"1529-7268"}],"subject":[],"published-other":{"date-parts":[[2025,10]]},"published":{"date-parts":[[2025,4,10]]}}}