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Ion channels from the mouse sperm plasma membrane in planar lipid bilayers

Published online by Cambridge University Press:  26 September 2008

Pedro Labarca
Affiliation:
Institute de Biotecnologia, Universidad Nacional Auténoma de México, and Centro de Estudios Cientificos de Santiago and Faculty of Sciences, Universidad de Chile.
Otilia Zapata
Affiliation:
Institute de Biotecnologia, Universidad Nacional Auténoma de México, and Centro de Estudios Cientificos de Santiago and Faculty of Sciences, Universidad de Chile.
Carmen Beltrán
Affiliation:
Institute de Biotecnologia, Universidad Nacional Auténoma de México, and Centro de Estudios Cientificos de Santiago and Faculty of Sciences, Universidad de Chile.
Alberto Darszon*
Affiliation:
Institute de Biotecnologia, Universidad Nacional Auténoma de México, and Centro de Estudios Cientificos de Santiago and Faculty of Sciences, Universidad de Chile.
*
Dr Alberto Darszon Darszon, Instituto de Biotecnologia, Apartado postal 510–3, Cuernavaca, Morelos, Mexico. Fax: 52-73-172388.

Summary

Fusion of purified mouse sperm plasma membranes to planar lipid bilayers resulted in the insertion of three ion channel types. They could be discerned on the basis of their selectivity, conductance, gating and voltage-dependent properties. The presence of a previously reported large, Ca2+-selective channel was confirmed. Here, it is reported that the Ca21-selective channel from mouse sperm plasmamembrane displayed a pNa+/Pk+ = 1.6 ± 0.2(n=4) and was blocked by micromolar concentrations of ruthenium red. Fusion yielded also a cation-selective channel (PNa+/Pk+ = 2.5±0.3, n=3) with a main open conductance substate of 103 pS and a smaller open substate of 51 PS(600mM K+cis/100 mM Na+trans). The channel inserted into bilayers in two stable fashions: a high-activity mode (open probability = 0.57 ± 0.02, n=3), and a low activity mode (open probability <1%, n=4). In high mode, the channel displayed bursting kinetics and burst length was voltage independent. In addition, a perfectly anion-selective channel, with a slope conductance of 83 PS (600KCI cis/100KCI trans), was identified. It displayed a high, nearly constant open probability (∼0.90)in the 0 to –80 mV range.

Type
Article
Copyright
Copyright © Cambridge University Press 1995

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References

Beltrán, C., Darszon, A., Labarca, P. & Liévano, A. (1994). A high-conductance multistate Ca2+ channel found in sea urchin and mouse spermatozoa. FEBS Lett. 338, 23–6.CrossRefGoogle ScholarPubMed
Bently, J.K., Garbers, D.L., Domino, S.E., Nolan, T.D.. & Van Dop, C. (1986). Spermatozoa contain a guanine nucleotide binding protein ADP ribosylated by pertussis toxin. Biochem. Biophys. Res. Commun. 138, 728–34.CrossRefGoogle Scholar
Coronado, R., Morrissette, J., Sukhareva, M.. & Vaughan, D.M. (1994). Structure and function of ryanodine receptors. Am. J. Physiol. (Cell Physiol.) 266, C1485–504.CrossRefGoogle ScholarPubMed
Cox, T.. & Peterson, R.N. (1989). Identification of calcium conducting channels in isolated boar sperm plasma membranes. Biochem. Biophys. Res. Commun. 161, 162–8.CrossRefGoogle ScholarPubMed
Cox, T., Campbell, P. & Peterson, R.N. (1991). Ion channels in boar sperm plasma membranes: characterization of a cation selective channel. Mol. Reprod. Dev. 30, 135–47.CrossRefGoogle ScholarPubMed
Darszon, A., Labarca, P., Beltrá, C., García-Soto, J..& Liévano, A. (1994). Sea urchin sperm: an ion channel reconstitution study case. Methods (a companion to Methods enzymol.) 6, 3750.CrossRefGoogle Scholar
Endo, Y., Lee, M.A. & Kopf, G.S. (1987). Evidence for a role for a guanine nucleotide binding regulatory protein in the zona pellucida induced sperm acrosome reaction. Dev. Biol. 119, 210–16.CrossRefGoogle ScholarPubMed
Florman, H.M. (1994). Sequential focal and global elevations of sperm intracellular Ca2+ are initiated by the zona pellucida during acrosomal exocytosis. Dev. Biol. 165, 152–64.CrossRefGoogle ScholarPubMed
Florman, H.M.. & Babcock, D.F. (1991). Progress toward understanding the molecular basis of capacitation. In Elements of Mammalian Ferlilization, Vol.1, Basic Concepts, Wassarman, P.M.. 105–32. Boca Raton, FL: CRC Press.Google Scholar
Florman, H., Corron, M.E., Kim, T.D.-H. & Babcock, D.F. (1992). Activation of voltage-dependent calcium channels of mammalian sperm is required for zona pellucida-induced acrosomal exocytosis. Dev. Biol. 152, 304–14.CrossRefGoogle ScholarPubMed
Guerrero, A. & Darszon, A. (1989).Evidence for the activation of two different Ca2+ channels during the egg jelly induced acrosome reaction of sea urchin sperm. J. Biol. Chem. 264, 19593–9.CrossRefGoogle ScholarPubMed
Hille, B. (1992). Ion channels of Excitable Membranes, 2nd edn. Sunderland, MA: Sinauer Associates.Google Scholar
Kopf, G.S., Woolkalis, M.J. & Gerton, G.L. (1986). Evidence for a guanine nucleotide-binding regulatory protein in invertebrate and mammalian sperm: identification by islet activating protein catalyzed ADP ribosylation and immunochemical methods. J. Biol. Chem. 261, 7327–31.CrossRefGoogle ScholarPubMed
Kostyuk, P., Akaike, N., Osipchuk, Y., Savchenko, A. & Shuba, Y. (1989). Gating and permeability properties of different types of Ca channels. Ann. NY Acad. Sci. 560, 6379.CrossRefGoogle Scholar
Liévano, A., Vega-Saenz de Miera, E.C. & Darszon, A. (1990). Ca2+ channels from the sea urchin sperm plasma membrane. J. Gen. Physiol. 95, 273–96.CrossRefGoogle ScholarPubMed
Ward, G.R. & Kopf, G. (1993). Molecular events mediating sperm activation. Dev. Biol. 158, 934.CrossRefGoogle ScholarPubMed
Wassarman, P.M. (1990). Profile of a mammalian sperm receptor. Development 108, 117.CrossRefGoogle ScholarPubMed
Weyand, I., Godde, M., Frings, S., Weiner, J., Müler, F., Altenhofen, W., Hatt, H. & Kaupp, B. (1994). Cooning and functional expression of a cyclic-nucleotide-gated channel from mammalian sperm. Nature 368, 859–63.CrossRefGoogle Scholar
Wistrom, C.A. & Meizel, S. (1993). Evidence supporting involvement of a unique human steriod receptor/Cl channel complex in the progesterone-initiated reaction. Dev. Biol. 159, 679–90.Google Scholar
Young, G.P.H., Koide, S.S., Goldstein, M. & Young, J.D.E. (1988). Isolation and partial characterization of an ion channel protein from human sperm membranes. Arch. Biochem. Biophys. 262, 491500.CrossRefGoogle ScholarPubMed