The microtubule plus-end-tracking protein CLIP-170 associates with the spermatid manchette and is essential for spermatogenesis

  1. Anna Akhmanova1,
  2. Anne-Laure Mausset-Bonnefont1,6,
  3. Wiggert van Cappellen2,
  4. Nanda Keijzer1,
  5. Casper C. Hoogenraad1,3,
  6. Tatiana Stepanova1,
  7. Ksenija Drabek1,
  8. Jacqueline van der Wees1,
  9. Mieke Mommaas4,
  10. Jos Onderwater4,
  11. Hans van der Meulen4,
  12. Marvin E. Tanenbaum5,
  13. Rene H. Medema5,
  14. Jos Hoogerbrugge2,
  15. Jan Vreeburg2,
  16. Evert-Jan Uringa2,
  17. J. Anton Grootegoed2,
  18. Frank Grosveld1, and
  19. Niels Galjart1,7
  1. 1Department of Cell Biology and Genetics, 2Department of Reproduction and Development, and 3Department of Neuroscience, Erasmus MC, 3000 DR Rotterdam, The Netherlands; 4Center for Electron Microscopy, Department of Molecular Cell Biology, Leiden University Medical Center, 2300 RA Leiden, The Netherlands; 5Department of Medical Oncology, University Medical Center, 3584 CG Utrecht, The Netherlands

Abstract

CLIP-170 is a microtubule “plus-end-tracking protein” implicated in the control of microtubule dynamics, dynactin localization, and the linking of endosomes to microtubules. To investigate the function of mouse CLIP-170, we generated CLIP-170 knockout and GFP-CLIP-170 knock-in alleles. Residual CLIP-170 is detected in lungs and embryos of homozygous CLIP-170 knockout mice, but not in other tissues and cell types, indicating that we have generated a hypomorphic mutant. Homozygous CLIP-170 knockout mice are viable and appear normal. However, male knockout mice are subfertile and produce sperm with abnormal heads. Using the knock-in mice, we followed GFP-CLIP-170 expression and behavior in dissected, live testis tubules. We detect plus-end-tracking GFP-CLIP-170 in spermatogonia. As spermatogenesis proceeds, GFP-CLIP-170 expression increases and the fusion protein strongly marks syncytia of differentiated spermatogonia and early prophase spermatocytes. Subsequently GFP-CLIP-170 levels drop, but during spermiogenesis (post-meiotic development), GFP-CLIP-170 accumulates again and is present on spermatid manchettes and centrosomes. Bleaching studies show that, as spermatogenesis progresses, GFP-CLIP-170 converts from a mobile plus-end-tracking protein to a relatively immobile protein. We propose that CLIP-170 has a structural function in the male germline, in particular in spermatid differentiation and sperm head shaping.

Keywords

Footnotes

  • Supplemental material is available at http://www.genesdev.org.

  • ↵6 Present address: INSERM U583, Institut des Neurosciences de Montpellier, Hôpital Saint-Eloi, 80 avenue Augustin Fliche, BP 74103, 34091 Montpellier Cedex 5, France.

  • ↵7 Corresponding author.

    ↵7 E-MAIL n.galjart{at}erasmusmc.nl; FAX 31-10-4089468.

    ↵7 Article and publication are at http://www.genesdev.org/cgi/doi/10.1101/gad.344505.

    • Accepted July 13, 2005.
    • Received March 20, 2005.
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  1. doi: 10.1101/gad.344505 Genes & Dev. 19: 2501-2515 Cold Spring Harbor Laboratory Press

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