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A new sperm-specific Na+/H+ Exchanger required for sperm motility and fertility

Nature Cell Biology volume 5, pages 1117–1122 (2003)Cite this article

Abstract

It has long been speculated that intracellular pH is a critical regulator of both invertebrate and vertebrate sperm motility1,2,3, and sodium–hydrogen exchange has been suggested as a mediator of such pHi regulation in various instances4,5. Two sodium–hydrogen exchangers (NHE1 and NHE5) are expressed in spermatozoa6. However, elimination of the NHE1 gene fails to cause infertility7, suggesting that normal sperm function is maintained in NHE1-null animals. Here, we used a functionally unbiased signal peptide trap screen to identify a novel sperm-specific NHE. The NHE contains 14 predicted transmembrane segments, including a potential voltage sensor and a consensus cyclic nucleotide-binding motif. Testis histology, sperm numbers and morphology were normal, but NHE-null males were completely infertile with severely diminished sperm motility. The addition of ammonium chloride, which elevates intracellular pH, partially rescued the motility and fertility defects. Surprisingly, cyclic AMP analogues almost completely rescued the motility and infertility phenotypes. The existence of this new sperm NHE provides an attractive contraceptive target, given its cell-specific expression and absolute requirement for fertility.

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Figure 1: A novel testis-specific NHE.
Figure 2: Western blot and immunolocalization of the sperm NHE.
Figure 3: Disruption of the sperm NHE gene.
Figure 4: Ammonium chloride and cAMP analogues rescue the motility defect of null spermatozoa.

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References

  1. Garbers, D.L., Watkins, H.D., Hansbrough, J.R., Smith, A. & Misono, K.S. The amino acid sequence and chemical synthesis of speract and of speract analogues. J. Biol. Chem. 257, 2734–2737 (1982).

    CAS  PubMed  Google Scholar 

  2. Babcock, D.F., Rufo, G.A., Jr & Lardy, H.A. Potassium-dependent increases in cytosolic pH stimulate metabolism and motility of mammalian sperm. Proc. Natl Acad. Sci. USA 80, 1327–1331 (1983).

    Article  CAS  Google Scholar 

  3. Carr, D.W. & Acott, T.S. Intracellular pH regulates bovine sperm motility and protein phosphorylation. Biol. Reprod. 41, 907–920 (1989).

    Article  CAS  Google Scholar 

  4. Garcia, M.A. & Meizel, S. Regulation of intracellular pH in capacitated human spermatozoa by a Na+/H+ exchanger. Mol. Reprod. Develop. 52, 189–195 (1999).

    Article  CAS  Google Scholar 

  5. Schackmann, R.W. & Chock, P.B. Alteration of intracellular [Ca2+] in sea urchin sperm by the egg peptide speract. Evidence that increased intracellular Ca2+ is coupled to Na+ entry and increased intracellular pH. J. Biol. Chem. 261, 8719–7828 (1986).

    CAS  Google Scholar 

  6. Woo, A.L., James, P.F. & Lingrel, J.B. Roles of the Na,K-ATPase α4 isoform and the Na+/H+ exchanger in sperm motility. Mol. Reprod. Develop. 62, 348–356 (2002).

    Article  CAS  Google Scholar 

  7. Bell, S.M. et al. Targeted disruption of the murine Nhe1 locus induces ataxia, growth retardation, and seizures. Am. J. Physiol. 276, C788–C795 (1999).

    Article  CAS  Google Scholar 

  8. Hansbrough, J.R. & Garbers, D.L. Speract. Purification and characterization of a peptide associated with eggs that activates spermatozoa. J. Biol. Chem. 256, 1447–1452 (1981).

    CAS  PubMed  Google Scholar 

  9. Moy, G.W. & Vacquier, V.D. Immunoperoxidase localization of bindin during the adhesion of sperm to sea urchin eggs. Curr. Top. Dev. Biol. 13, 31–44 (1979).

    Article  CAS  Google Scholar 

  10. Garbers, D.L. Guanylyl cyclase receptors and their endocrine, paracrine, and autocrine ligands. Cell 71, 1–4 (1992).

    Article  CAS  Google Scholar 

  11. Mengerink, K.J., Moy, G.W. & Vacquier, V.D. suREJ3, a polycystin-1 protein, is cleaved at the GPS domain and localizes to the acrosomal region of sea urchin sperm. J. Biol. Chem. 277, 943–948 (2002).

    Article  CAS  Google Scholar 

  12. Klein, R.D., Gu, Q., Goddard, A. & Rosenthal, A. Selection for genes encoding secreted proteins and receptors. Proc. Natl Acad. Sci. USA 93, 7108–7113 (1996).

    Article  CAS  Google Scholar 

  13. Jacobs, K.A. et al. A genetic selection for isolating cDNAs encoding secreted proteins. Gene 198, 289–296 (1997).

    Article  CAS  Google Scholar 

  14. Quill, T.A., Ren, D., Clapham, D.E. & Garbers, D.L. A voltage-gated ion channel expressed specifically in spermatozoa. Proc. Natl Acad. Sci. USA 98, 12527–12531 (2001).

    Article  CAS  Google Scholar 

  15. Counillon, L. & Pouyssegur, J. The expanding family of eucaryotic Na(+)/H(+) exchangers. J. Biol. Chem. 275, 1–4 (2000).

    Article  CAS  Google Scholar 

  16. Numata, M. & Orlowski, J. Molecular cloning and characterization of a novel (Na+,K+)/H+ exchanger localized to the trans-Golgi network. J. Biol. Chem. 276, 17387–17394 (2001).

    Article  CAS  Google Scholar 

  17. Goyal, S., Vanden Heuvel, G. & Aronson, P.S. Renal expression of novel Na+/H+ exchanger isoform NHE8. Am. J. Physiol. Renal Physiol. 284, F467–F473 (2003).

    Article  CAS  Google Scholar 

  18. Lee, H.C. The voltage-sensitive Na+/H+ exchange in sea urchin spermatozoa flagellar membrane vesicles studied with an entrapped pH probe. J. Biol. Chem. 260, 10794–10799 (1985).

    CAS  PubMed  Google Scholar 

  19. Ren, D. et al. A sperm ion channel required for sperm motility and male fertility. Nature 413, 603–609 (2001).

    Article  CAS  Google Scholar 

  20. Watanabe, D. et al. Molecular cloning of a novel Ca(2+)-binding protein (calmegin) specifically expressed during male meiotic germ cell development. J. Biol. Chem. 269, 7744–7749 (1994).

    CAS  PubMed  Google Scholar 

  21. Ikawa, M. et al. The putative chaperone calmegin is required for sperm fertility. Nature 387, 607–611 (1997).

    Article  CAS  Google Scholar 

  22. Roos, A. & Boron, W.F. Intracellular pH. Physiol. Rev. 61, 296–434 (1981).

    Article  CAS  Google Scholar 

  23. Howlett, S.K. & Bolton, V.N. Sequence and regulation of morphological and molecular events during the first cell cycle of mouse embryogenesis. J. Embryol. Experiment Morphol. 87, 175–206 (1985).

    CAS  Google Scholar 

  24. Fraser, L.R. In vitro capacitation and fertilization. Methods Enzymol. 225, 239–253 (1993).

    Article  CAS  Google Scholar 

  25. Si, Y. & Olds-Clarke, P. Evidence for the involvement of calmodulin in mouse sperm capacitation. Biol. Reprod. 62, 1231–1239 (2000).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank E. Lummus for technical assistance, R.E. Hammer for production of the animal chimaeras, A. Carlson and D. Bobcock for help with pHi measurements, and J. Pouyssegur for assisting in the determination of NHE expression and activity in NHE-deficient cells. We also appreciate the use of the Molecular Pathology Core Laboratory and Molecular and Cellular Imaging Facility at the University of Texas Southwestern Medical Center. This work was supported by the Howard Hughes Medical Institute, the Cecil H. & Ida Green Center for Reproductive Biology Sciences and grant HD 36022 from the National Institutes of Health.

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Authors and Affiliations

  1. Cecil H. & Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, 75390-9051, TX

    Dan Wang, Shelby M. King, Timothy A. Quill, Lynda K. Doolittle & David L. Garbers

  2. Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, 75390-9051, TX

    Dan Wang, Lynda K. Doolittle & David L. Garbers

  3. Department of Pharmacology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, 75390-9051, TX

    Dan Wang, Shelby M. King, Timothy A. Quill, Lynda K. Doolittle & David L. Garbers

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  1. Dan Wang
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Correspondence to David L. Garbers.

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Wang, D., King, S., Quill, T. et al. A new sperm-specific Na+/H+ Exchanger required for sperm motility and fertility. Nat Cell Biol 5, 1117–1122 (2003). https://doi.org/10.1038/ncb1072

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