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Flagellar Ca\n                    <jats:sup>2+<\/jats:sup>\n                    signaling nanodomains, organized by multi-subunit CatSper calcium channel complexes, are pivotal for sperm migration in the female tract, implicating CatSper-dependent mechanisms in sperm selection. Here using biochemical and pharmacological studies, we demonstrate that CatSper1 is an O-linked glycosylated protein, undergoing capacitation-induced processing dependent on Ca\n                    <jats:sup>2+<\/jats:sup>\n                    and phosphorylation cascades. CatSper1 processing correlates with protein tyrosine phosphorylation (pY) development in sperm cells capacitated in vitro and in vivo. Using 3D in situ molecular imaging and ANN-based automatic detection of sperm distributed along the cleared female tract, we demonstrate that spermatozoa past the utero-tubal junction possess the intact CatSper1 signals. Together, we reveal that fertilizing mouse spermatozoa in situ are characterized by intact CatSper channel, lack of pY, and reacted acrosomes. These findings provide molecular insight into sperm selection for successful fertilization in the female reproductive tract.\n                  <\/jats:p>","DOI":"10.7554\/elife.62043","type":"journal-article","created":{"date-parts":[[2020,10,20]],"date-time":"2020-10-20T08:00:19Z","timestamp":1603180819000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.62043","source":"Crossref","is-referenced-by-count":60,"title":["3D in situ imaging of the female reproductive tract reveals molecular signatures of fertilizing spermatozoa in mice"],"prefix":"10.7554","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1053-4025","authenticated-orcid":true,"given":"Lukas","family":"Ded","sequence":"first","affiliation":[{"name":"Department of Cellular & Molecular Physiology, Yale School of Medicine, New Haven, United States"},{"name":"Laboratory of Reproductive Biology, Institute of Biotechnology, Czech Academy of Sciences, BIOCEV, Vestec, Czech 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