3D in situ imaging of female reproductive tract reveals molecular signatures of fertilizing spermatozoa in mice
- Yale School of Medicine, United States
- Boston Children's Hospital, United States
Abstract
Out of millions of ejaculated sperm, only a few reach the fertilization site in mammals. Flagellar Ca2+ 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 Ca2+ 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 all spermatozoa past the UTJ possess 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.
Data availability
All data generated or analysed during this study are included in the manuscript, supplementary and source data files.
Article and author information
Author details
Funding
National Institutes of Health (R01HD096745)
- Jean-Ju Chung
Yale School of Medicine (Start-up funds)
- Jean-Ju Chung
Yale University (a Yale Goodman-Gilman ScholarAward-2015)
- Jean-Ju Chung
Male Contraceptive Initiative (Postdoctoral fellowship)
- Jae Yeon Hwang
Czech Science Foundation (GJ20-17403Y)
- Lukas Ded
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All the mice were treated in accordance with guidelines approved by Yale (20079) Animal Care and Use Committees (IACUC).
Copyright
© 2020, Ded et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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3D in situ imaging of female reproductive tract reveals molecular signatures of fertilizing spermatozoa in mice
eLife 9:e62043.
https://doi.org/10.7554/eLife.62043
