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. 1985 Mar;82(5):1460–1464. doi: 10.1073/pnas.82.5.1460

Calcium channel antagonists inhibit the acrosome reaction and bind to plasma membranes of sea urchin sperm.

T Kazazoglou, R W Schackmann, M Fosset, B M Shapiro
PMCID: PMC397282  PMID: 3856274

Abstract

As a prerequisite to fertilization, sea urchin sperm undergo an acrosome reaction that is mediated in part by increased permeability to Ca2+, with an attendant rapid, massive intracellular Ca2+ accumulation. The acrosome reaction is inhibited by Ca2+ channel antagonists, including verapamil, D600, and dihydropyridines such as nitrendipine, nimodipine, and nisoldipine. To examine the interaction of Ca2+ antagonists with sperm, a plasma membrane preparation enriched for Na+,K+-ATPase was isolated from sea urchin sperm. These plasma membranes specifically bound [3H]nitrendipine and [3H]verapamil at concentrations similar to those that inhibit the acrosome reaction. The binding of verapamil was sigmoidal and half-maximal at 1 microM. There was a high specificity in the binding interaction, since by competition binding verapamil, (-)-D600, and (+)-D600 had different relative Kd values, 11, 2.5, and 0.5 microM, respectively. These data suggest that sperm mediate the Ca2+ influx required for induction of the acrosome reaction via Ca2+ channels with properties similar, but not identical, to those of other excitable tissues.

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Selected References

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  1. APPEL W. [(N-methyl-N-homoveratryl)-gamma-aminopropyl] -3,4-dimethoxyphenylacetonitrile, its detection in biological materials and its behavior in blood]. Arzneimittelforschung. 1962 Jun;12:562–566. [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Cantino M. E., Schackmann R. W., Johnson D. E. Changes in subcellular elemental distributions accompanying the acrosome reaction in sea urchin sperm. J Exp Zool. 1983 May;226(2):255–268. doi: 10.1002/jez.1402260211. [DOI] [PubMed] [Google Scholar]
  4. Christen R., Schackmann R. W., Shapiro B. M. Interactions between sperm and sea urchin egg jelly. Dev Biol. 1983 Jul;98(1):1–14. doi: 10.1016/0012-1606(83)90330-5. [DOI] [PubMed] [Google Scholar]
  5. Collins F., Epel D. The role of calcium ions in the acrosome reaction of sea urchin sperm: regulation of exocytosis. Exp Cell Res. 1977 Apr;106(1):211–222. doi: 10.1016/0014-4827(77)90258-0. [DOI] [PubMed] [Google Scholar]
  6. Cross N. L. Isolation and electrophoretic characterization of the plasma membrane of sea-urchin sperm. J Cell Sci. 1983 Jan;59:13–25. doi: 10.1242/jcs.59.1.13. [DOI] [PubMed] [Google Scholar]
  7. Curtis B. M., Catterall W. A. Solubilization of the calcium antagonist receptor from rat brain. J Biol Chem. 1983 Jun 25;258(12):7280–7283. [PubMed] [Google Scholar]
  8. DePover A., Lee S. W., Matlib M. A., Whitmer K., Davis B. A., Powell T., Schwartz A. [3H]nimodipine specific binding to cardiac myocytes and subcellular fractions. Biochem Biophys Res Commun. 1983 May 31;113(1):185–191. doi: 10.1016/0006-291x(83)90449-7. [DOI] [PubMed] [Google Scholar]
  9. Decker G. L., Joseph D. B., Lennarz W. J. A study of factors involved in induction of the acrosomal reaction in sperm of the sea urchin, Arbacia punctulata. Dev Biol. 1976 Oct 1;53(1):115–125. doi: 10.1016/0012-1606(76)90213-x. [DOI] [PubMed] [Google Scholar]
  10. Ehlert F. J., Itoga E., Roeske W. R., Yamamura H. I. The interaction of [3H]nitrendipine with receptors for calcium antagonists in the cerebral cortex and heart of rats. Biochem Biophys Res Commun. 1982 Feb 11;104(3):937–943. doi: 10.1016/0006-291x(82)91339-0. [DOI] [PubMed] [Google Scholar]
  11. Fosset M., Jaimovich E., Delpont E., Lazdunski M. [3H]nitrendipine labelling of the Ca2+ channel in skeletal muscle. Eur J Pharmacol. 1982 Dec 17;86(1):141–142. doi: 10.1016/0014-2999(82)90414-9. [DOI] [PubMed] [Google Scholar]
  12. Fosset M., Jaimovich E., Delpont E., Lazdunski M. [3H]nitrendipine receptors in skeletal muscle. J Biol Chem. 1983 May 25;258(10):6086–6092. [PubMed] [Google Scholar]
  13. Gache C., Rossi B., Lazdunski M. (Na+, K+)-activated adenosinetriphosphatase of axonal membranes, cooperativity and control. Steady-state analysis. Eur J Biochem. 1976 May 17;65(1):293–306. doi: 10.1111/j.1432-1033.1976.tb10417.x. [DOI] [PubMed] [Google Scholar]
  14. Galizzi J. P., Fosset M., Lazdunski M. [3H] verapamil binding sites in skeletal muscle transverse tubule membranes. Biochem Biophys Res Commun. 1984 Jan 13;118(1):239–245. doi: 10.1016/0006-291x(84)91092-1. [DOI] [PubMed] [Google Scholar]
  15. Garbers D. L. The elevation of cyclic AMP concentrations in flagella-less sea urchin sperm heads. J Biol Chem. 1981 Jan 25;256(2):620–624. [PubMed] [Google Scholar]
  16. Gould R. J., Murphy K. M., Snyder S. H. [3H]nitrendipine-labeled calcium channels discriminate inorganic calcium agonists and antagonists. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3656–3660. doi: 10.1073/pnas.79.11.3656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Grover A. K., Kwan C. Y., Luchowski E., Daniel E. E., Triggle D. J. Subcellular distribution of [3H]nitrendipine binding in smooth muscle. J Biol Chem. 1984 Feb 25;259(4):2223–2226. [PubMed] [Google Scholar]
  18. Hagiwara S., Byerly L. Calcium channel. Annu Rev Neurosci. 1981;4:69–125. doi: 10.1146/annurev.ne.04.030181.000441. [DOI] [PubMed] [Google Scholar]
  19. Kazazoglou T., Schmid A., Renaud J. F., Lazdunski M. Ontogenic appearance of Ca2+ channels characterized as binding sites for nitrendipine during development of nervous, skeletal and cardiac muscle systems in the rat. FEBS Lett. 1983 Nov 28;164(1):75–79. doi: 10.1016/0014-5793(83)80022-2. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Langer G. A. Heart: excitation-contraction coupling. Annu Rev Physiol. 1973;35:55–86. doi: 10.1146/annurev.ph.35.030173.000415. [DOI] [PubMed] [Google Scholar]
  23. Mason T. L., Poyton R. O., Wharton D. C., Schatz G. Cytochrome c oxidase from bakers' yeast. I. Isolation and properties. J Biol Chem. 1973 Feb 25;248(4):1346–1354. [PubMed] [Google Scholar]
  24. Renaud J. F., Kazazoglou T., Schmid A., Romey G., Lazdunski M. Differentiation of receptor sites for [3H]nitrendipine in chick hearts and physiological relation to the slow Ca2+ channel and to excitation-contraction coupling. Eur J Biochem. 1984 Mar 15;139(3):673–681. doi: 10.1111/j.1432-1033.1984.tb08056.x. [DOI] [PubMed] [Google Scholar]
  25. Schackmann R. W., Christen R., Shapiro B. M. Membrane potential depolarization and increased intracellular pH accompany the acrosome reaction of sea urchin sperm. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6066–6070. doi: 10.1073/pnas.78.10.6066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schackmann R. W., Eddy E. M., Shapiro B. M. The acrosome reaction of Strongylocentrotus purpuratus sperm. Ion requirements and movements. Dev Biol. 1978 Aug;65(2):483–495. doi: 10.1016/0012-1606(78)90043-x. [DOI] [PubMed] [Google Scholar]
  27. Schackmann R. W., Shapiro B. M. A partial sequence of ionic changes associated with the acrosome reaction of Strongylocentrotus purpuratus. Dev Biol. 1981 Jan 15;81(1):145–154. doi: 10.1016/0012-1606(81)90357-2. [DOI] [PubMed] [Google Scholar]
  28. Shapiro B. M., Schackmann R. W., Gabel C. A. Molecular approaches to the study of fertilization. Annu Rev Biochem. 1981;50:815–843. doi: 10.1146/annurev.bi.50.070181.004123. [DOI] [PubMed] [Google Scholar]
  29. Talbot P., Summers R. G., Hylander B. L., Keough E. M., Franklin L. E. The role of calcium in the acrosome reaction: an analysis using ionophore A23187. J Exp Zool. 1976 Dec;198(3):383–392. doi: 10.1002/jez.1401980312. [DOI] [PubMed] [Google Scholar]
  30. Tsien R. W. Calcium channels in excitable cell membranes. Annu Rev Physiol. 1983;45:341–358. doi: 10.1146/annurev.ph.45.030183.002013. [DOI] [PubMed] [Google Scholar]
  31. Williams L. T., Jones L. R. Specific binding of the calcium antagonist [3H]nitrendipine to subcellular fractions isolated from canine myocardium. Evidence for high affinity binding to ryanodine-sensitive sarcoplasmic reticulum vesicles. J Biol Chem. 1983 May 10;258(9):5344–5347. [PubMed] [Google Scholar]

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