DNALI1 interacts with the MEIG1/PACRG complex within the manchette and is required for proper sperm flagellum assembly in mice

  1. Yi Tian Yap
  2. Wei Li
  3. Qian Huang
  4. Qi Zhou
  5. David Zhang
  6. Yi Sheng
  7. Ljljiana Mladenovic-Lucas
  8. Siu-Pok Yee
  9. Kyle E Orwig
  10. James G Granneman
  11. David C Williams Jr
  12. Rex Hess
  13. Aminata Toure
  14. Zhibing Zhang  Is a corresponding author
  1. Wayne State University, United States
  2. Wuhan University of Science and Technology, China
  3. College of William and Mary, United States
  4. University of Pittsburgh, United States
  5. University of Connecticut Health Center, United States
  6. University of North Carolina at Chapel Hill, United States
  7. University of Illinois Urbana-Champaign, United States
  8. 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

  1. Yi Tian Yap

    Department of Physiology, Wayne State University, Detroit, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6448-2748
  2. Wei Li

    Department of Physiology, Wayne State University, Detroit, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Qian Huang

    Department of Occupational and Environmental Medicine, Wuhan University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6836-5135
  4. Qi Zhou

    Department of Occupational and Environmental Medicine, Wuhan University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  5. David Zhang

    College of William and Mary, Williamsburg, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Yi Sheng

    Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh, Pittsburgh, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Ljljiana Mladenovic-Lucas

    Center for Molecular Medicine and Genetics, Wayne State University, Detroit, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Siu-Pok Yee

    Department of Cell Biology, University of Connecticut Health Center, Farmington, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Kyle E Orwig

    Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh, Pittsburgh, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. James G Granneman

    Center for Molecular Medicine and Genetics, Wayne State University, Detroit, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7013-6630
  11. David C Williams Jr

    Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6536-4038
  12. Rex Hess

    Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Aminata Toure

    Institute for Advanced Biosciences, Université Grenoble Alpes, INSERM U1209, CNRS UMR 5309, Grenoble, France
    Competing interests
    The authors declare that no competing interests exist.
  14. Zhibing Zhang

    Department of Physiology, Wayne State University, Detroit, United States
    For correspondence
    gn6075@wayne.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8615-4478

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.

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).

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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  1. Yi Tian Yap
  2. Wei Li
  3. Qian Huang
  4. Qi Zhou
  5. David Zhang
  6. Yi Sheng
  7. Ljljiana Mladenovic-Lucas
  8. Siu-Pok Yee
  9. Kyle E Orwig
  10. James G Granneman
  11. David C Williams Jr
  12. Rex Hess
  13. Aminata Toure
  14. Zhibing Zhang
(2023)
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

Share this article

https://doi.org/10.7554/eLife.79620

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