DNALI1 interacts with the MEIG1/PACRG complex within the manchette and is required for proper sperm flagellum assembly in mice
- Wayne State University, United States
- Wuhan University of Science and Technology, China
- College of William and Mary, United States
- University of Pittsburgh, United States
- University of Connecticut Health Center, United States
- University of North Carolina at Chapel Hill, United States
- University of Illinois Urbana-Champaign, United States
- Université Grenoble Alpes, INSERM U1209, CNRS UMR 5309, France
Abstract
The manchette is a transient and unique structure present in elongating spermatids and required for proper differentiation of the germ cells during spermatogenesis. Previous work indicated that the MEIG1/PACRG complex locates in the manchette and is involved in the transport of cargos, such as SPAG16L, to build the sperm flagellum. Here using co-immunoprecipitation and pull-down approaches in various cell systems, we established that DNALI1, an axonemal component originally cloned from Chlamydomonas reinhardtii, recruits and stabilizes PACRG and we confirm in vivo, the co-localization of DNALI1 and PACRG in the manchette by immunofluorescence of elongating murine spermatids. We next generated mice with a specific deficiency of DNALI1 in male germ cells, and observed a dramatic reduction of the sperm cells, which results in male infertility. In addition, we observed that the majority of the sperm cells exhibited abnormal morphology including misshapen heads, bent tails, enlarged midpiece, discontinuous accessory structure, emphasizing the importance of DNALI1 in sperm differentiation. Examination of testis histology confirmed impaired spermiogenesis in the mutant mice. Importantly, while testicular levels of MEIG1, PACRG and SPAG16L proteins were unchanged in the Dnali1 mutant mice, their localization within the manchette was greatly affected, indicating that DNALI1 is required for the formation of the MEIG1/PACRG complex within the manchette. Interestingly, in contrast to MEIG1 and PACRG-deficient mice, the DNALI1-deficient mice also showed impaired sperm spermiation/individualization, suggesting additional functions beyond its involvement in the manchette structure. Overall, our work identifies DNALI1 as a protein required for sperm development.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for Figures 2, 3, 5 and 9
Article and author information
Author details
David C Williams Jr
Funding
Wayne State University Startup fund
- Zhibing Zhang
Wayne State University Research Fund
- Zhibing Zhang
Male Contraceptive Initiative fellowship
- Yi Tian Yap
Male Contraceptive Initiative pilot award
- Zhibing Zhang
National institute of child health and human development
- Zhibing Zhang
National Institute of Diabetes and Digestive and Kidney Diseases
- James G Granneman
National Institute of Diabetes and Digestive and Kidney Diseases
- David C Williams Jr
Agence Nationale pour la Recherche
- Aminata Toure
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Jean-Ju Chung, Yale University, United States
Ethics
Animal experimentation: All animal research was executed in compliance with the guidelines of the Wayne State University Institutional Animal Care with the Program Advisory Committee (Protocol number: 18-02-0534).
Version history
- Received: April 20, 2022
- Preprint posted: April 30, 2022 (view preprint)
- Accepted: March 12, 2023
- Accepted Manuscript published: April 21, 2023 (version 1)
- Version of Record published: May 15, 2023 (version 2)
Copyright
© 2023, Yap 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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DNALI1 interacts with the MEIG1/PACRG complex within the manchette and is required for proper sperm flagellum assembly in mice
eLife 12:e79620.
https://doi.org/10.7554/eLife.79620
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