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. 1990 Feb 1;95(2):273–296. doi: 10.1085/jgp.95.2.273

Ca2+ channels from the sea urchin sperm plasma membrane

PMCID: PMC2216321  PMID: 2155281

Abstract

Ca2+ influx across the sea urchin sperm plasma membrane is a necessary step during the egg jelly-induced acrosome reaction. There is pharmacological evidence for the involvement of Ca2+ channels in this influx, but their presence has not been directly demonstrated because of the small size of this cell. Sea urchin sperm Ca2+ channels are being studied by fusing isolated plasma membranes into planar lipid bilayers. With this strategy, a Ca2+ channel has been detected with the following characteristics: (a) the channel exhibits a high mainstate conductance (gamma MS) of 172 pS in 50 mM CaCl2 solutions with voltage- dependent decaying to smaller conductance states at negative Em; (b) the channel is blocked by millimolar concentrations of Cd2+, Co2+, and La3+, which also inhibit the egg jelly-induced acrosome reaction; (c) the gamma MS conductance sequence for the tested divalent cations is the following: Ba2+ greater than Sr2+ greater than Ca2+; and (d) the channel discriminates poorly for divalent over monovalent cations (PCa/PNa = 5.9). The sperm Ca2+ channel gamma MS rectifies in symmetrical 10 mM CaCl2, having a maximal slope conductance value of 94 pS at +100 mV applied to the cis side of the bilayer. Under these conditions, a different single-channel activity of lesser conductance became apparent above the gamma MS current at positive membrane potentials. Also in 10 mM Ca2+ solutions, Mg2+ permeates through the main channel when added to the cis side with a PCa/PMg = 2.9, while it blocks when added to the trans side. In 50 mM Ca2+ solutions, the gamma MS open probability has values of 1.0 at voltages more positive than - 40 mV and decreases at more negatives potentials, following a Boltzmann function with an E0.5 = -72 mV and an apparent gating charge value of 3.9. These results describe a novel Ca2(+)-selective channel, and suggest that the main channel works as a single multipore assembly.

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Selected References

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  1. Bell J. E., Miller C. Effects of phospholipid surface charge on ion conduction in the K+ channel of sarcoplasmic reticulum. Biophys J. 1984 Jan;45(1):279–287. doi: 10.1016/S0006-3495(84)84154-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benham C. D., Tsien R. W. A novel receptor-operated Ca2+-permeable channel activated by ATP in smooth muscle. Nature. 1987 Jul 16;328(6127):275–278. doi: 10.1038/328275a0. [DOI] [PubMed] [Google Scholar]
  3. Bezanilla F. A high capacity data recording device based on a digital audio processor and a video cassette recorder. Biophys J. 1985 Mar;47(3):437–441. doi: 10.1016/S0006-3495(85)83935-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Christen R., Schackmann R. W., Shapiro B. M. Metabolism of sea urchin sperm. Interrelationships between intracellular pH, ATPase activity, and mitochondrial respiration. J Biol Chem. 1983 May 10;258(9):5392–5399. [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. Cox T., Peterson R. N. Identification of calcium conducting channels in isolated boar sperm plasma membranes. Biochem Biophys Res Commun. 1989 May 30;161(1):162–168. doi: 10.1016/0006-291x(89)91575-1. [DOI] [PubMed] [Google Scholar]
  7. Coyne M. D., Dagan D., Levitan I. B. Calcium and barium permeable channels from Aplysia nervous system reconstituted in lipid bilayers. J Membr Biol. 1987;97(3):205–213. doi: 10.1007/BF01869223. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Darszon A., Gould M., De De La Torre L., Vargas I. Response of isolated sperm plasma membranes from sea urchin to egg jelly. Eur J Biochem. 1984 Nov 2;144(3):515–522. doi: 10.1111/j.1432-1033.1984.tb08496.x. [DOI] [PubMed] [Google Scholar]
  10. Domino S. E., Garbers D. L. The fucose-sulfate glycoconjugate that induces an acrosome reaction in spermatozoa stimulates inositol 1,4,5-trisphosphate accumulation. J Biol Chem. 1988 Jan 15;263(2):690–695. [PubMed] [Google Scholar]
  11. Eisenman G., Dani J. A. An introduction to molecular architecture and permeability of ion channels. Annu Rev Biophys Biophys Chem. 1987;16:205–226. doi: 10.1146/annurev.bb.16.060187.001225. [DOI] [PubMed] [Google Scholar]
  12. Fox J. A. Ion channel subconductance states. J Membr Biol. 1987;97(1):1–8. doi: 10.1007/BF01869609. [DOI] [PubMed] [Google Scholar]
  13. Garbers D. L., Hardman J. G. Factors released from sea urchin eggs affect cyclic nucleotide metabolism in sperm. Nature. 1975 Oct 23;257(5528):677–678. doi: 10.1038/257677a0. [DOI] [PubMed] [Google Scholar]
  14. García-Soto J., Darszon A. High pH-induced acrosome reaction and Ca2+ uptake in sea urchin sperm suspended in Na+-free seawater. Dev Biol. 1985 Aug;110(2):338–345. doi: 10.1016/0012-1606(85)90093-4. [DOI] [PubMed] [Google Scholar]
  15. García-Soto J., González-Martínez M., de De la Torre L., Darszon A. Internal pH can regulate Ca2+ uptake and the acrosome reaction in sea urchin sperm. Dev Biol. 1987 Mar;120(1):112–120. doi: 10.1016/0012-1606(87)90109-6. [DOI] [PubMed] [Google Scholar]
  16. Geletyuk V. I., Kazachenko V. N. Single Cl- channels in molluscan neurones: multiplicity of the conductance states. J Membr Biol. 1985;86(1):9–15. doi: 10.1007/BF01871605. [DOI] [PubMed] [Google Scholar]
  17. Golowasch J., Kirkwood A., Miller C. Allosteric effects of Mg2+ on the gating of Ca2+-activated K+ channels from mammalian skeletal muscle. J Exp Biol. 1986 Sep;124:5–13. doi: 10.1242/jeb.124.1.5. [DOI] [PubMed] [Google Scholar]
  18. González-Martínez M., Darszon A. A fast transient hyperpolarization occurs during the sea urchin sperm acrosome reaction induced by egg jelly. FEBS Lett. 1987 Jun 29;218(2):247–250. doi: 10.1016/0014-5793(87)81055-4. [DOI] [PubMed] [Google Scholar]
  19. Guerrero A., Darszon A. Egg jelly triggers a calcium influx which inactivates and is inhibited by calmodulin antagonists in the sea urchin sperm. Biochim Biophys Acta. 1989 Mar 27;980(1):109–116. doi: 10.1016/0005-2736(89)90206-x. [DOI] [PubMed] [Google Scholar]
  20. Guerrero A., Darszon A. Evidence for the activation of two different Ca2+ channels during the egg jelly-induced acrosome reaction of sea urchin sperm. J Biol Chem. 1989 Nov 25;264(33):19593–19599. [PubMed] [Google Scholar]
  21. Guerrero A., Sánchez J. A., Darszon A. Single-channel activity in sea urchin sperm revealed by the patch-clamp technique. FEBS Lett. 1987 Aug 17;220(2):295–298. doi: 10.1016/0014-5793(87)80833-5. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Horie M., Irisawa H., Noma A. Voltage-dependent magnesium block of adenosine-triphosphate-sensitive potassium channel in guinea-pig ventricular cells. J Physiol. 1987 Jun;387:251–272. doi: 10.1113/jphysiol.1987.sp016572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Iijima T., Ciani S., Hagiwara S. Effects of the external pH on Ca channels: experimental studies and theoretical considerations using a two-site, two-ion model. Proc Natl Acad Sci U S A. 1986 Feb;83(3):654–658. doi: 10.1073/pnas.83.3.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kazachenko V. N., Geletyuk V. I. The potential-dependent K+ channel in molluscan neurones is organized in a cluster of elementary channels. Biochim Biophys Acta. 1984 Jun 13;773(1):132–142. doi: 10.1016/0005-2736(84)90558-3. [DOI] [PubMed] [Google Scholar]
  26. Kazazoglou T., Schackmann R. W., Fosset M., Shapiro B. M. Calcium channel antagonists inhibit the acrosome reaction and bind to plasma membranes of sea urchin sperm. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1460–1464. doi: 10.1073/pnas.82.5.1460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Konnerth A., Lux H. D., Morad M. Proton-induced transformation of calcium channel in chick dorsal root ganglion cells. J Physiol. 1987 May;386:603–633. doi: 10.1113/jphysiol.1987.sp016553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Krouse M. E., Schneider G. T., Gage P. W. A large anion-selective channel has seven conductance levels. Nature. 1986 Jan 2;319(6048):58–60. doi: 10.1038/319058a0. [DOI] [PubMed] [Google Scholar]
  29. Lee H. C. A membrane potential-sensitive Na+-H+ exchange system in flagella isolated from sea urchin spermatozoa. J Biol Chem. 1984 Dec 25;259(24):15315–15319. [PubMed] [Google Scholar]
  30. Lee H. C., Garbers D. L. Modulation of the voltage-sensitive Na+/H+ exchange in sea urchin spermatozoa through membrane potential changes induced by the egg peptide speract. J Biol Chem. 1986 Dec 5;261(34):16026–16032. [PubMed] [Google Scholar]
  31. 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. 1985 Sep 5;260(19):10794–10799. [PubMed] [Google Scholar]
  32. Lievano A., Sanchez J. A., Darszon A. Single-channel activity of bilayers derived from sea urchin sperm plasma membranes at the tip of a patch-clamp electrode. Dev Biol. 1985 Nov;112(1):253–257. doi: 10.1016/0012-1606(85)90140-x. [DOI] [PubMed] [Google Scholar]
  33. MacDermott A. B., Mayer M. L., Westbrook G. L., Smith S. J., Barker J. L. NMDA-receptor activation increases cytoplasmic calcium concentration in cultured spinal cord neurones. 1986 May 29-Jun 4Nature. 321(6069):519–522. doi: 10.1038/321519a0. [DOI] [PubMed] [Google Scholar]
  34. Mauro A., Blake M., Labarca P. Voltage gating of conductance in lipid bilayers induced by porin from outer membrane of Neisseria gonorrhoeae. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1071–1075. doi: 10.1073/pnas.85.4.1071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Miller C., Racker E. Ca++-induced fusion of fragmented sarcoplasmic reticulum with artificial planar bilayers. J Membr Biol. 1976;30(3):283–300. doi: 10.1007/BF01869673. [DOI] [PubMed] [Google Scholar]
  36. Miller C., White M. M. Dimeric structure of single chloride channels from Torpedo electroplax. Proc Natl Acad Sci U S A. 1984 May;81(9):2772–2775. doi: 10.1073/pnas.81.9.2772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nowak L., Bregestovski P., Ascher P., Herbet A., Prochiantz A. Magnesium gates glutamate-activated channels in mouse central neurones. Nature. 1984 Feb 2;307(5950):462–465. doi: 10.1038/307462a0. [DOI] [PubMed] [Google Scholar]
  38. Pietrobon D., Prod'hom B., Hess P. Conformational changes associated with ion permeation in L-type calcium channels. Nature. 1988 May 26;333(6171):373–376. doi: 10.1038/333373a0. [DOI] [PubMed] [Google Scholar]
  39. Prod'hom B., Pietrobon D., Hess P. Direct measurement of proton transfer rates to a group controlling the dihydropyridine-sensitive Ca2+ channel. Nature. 1987 Sep 17;329(6136):243–246. doi: 10.1038/329243a0. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. 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]
  42. 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]
  43. Smith J. S., Coronado R., Meissner G. Sarcoplasmic reticulum contains adenine nucleotide-activated calcium channels. Nature. 1985 Aug 1;316(6027):446–449. doi: 10.1038/316446a0. [DOI] [PubMed] [Google Scholar]
  44. Smith J. S., Coronado R., Meissner G. Single channel measurements of the calcium release channel from skeletal muscle sarcoplasmic reticulum. Activation by Ca2+ and ATP and modulation by Mg2+. J Gen Physiol. 1986 Nov;88(5):573–588. doi: 10.1085/jgp.88.5.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Smith J. S., Imagawa T., Ma J., Fill M., Campbell K. P., Coronado R. Purified ryanodine receptor from rabbit skeletal muscle is the calcium-release channel of sarcoplasmic reticulum. J Gen Physiol. 1988 Jul;92(1):1–26. doi: 10.1085/jgp.92.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Squire L. G., Petersen O. H. Modulation of Ca2+- and voltage-activated K+ channels by internal Mg2+ in salivary acinar cells. Biochim Biophys Acta. 1987 May 29;899(2):171–175. doi: 10.1016/0005-2736(87)90397-x. [DOI] [PubMed] [Google Scholar]
  47. Toowicharanont P., Shapiro B. M. Regional differentiation of the sea urchin sperm plasma membrane. J Biol Chem. 1988 May 15;263(14):6877–6883. [PubMed] [Google Scholar]
  48. Trimmer J. S., Trowbridge I. S., Vacquier V. D. Monoclonal antibody to a membrane glycoprotein inhibits the acrosome reaction and associated Ca2+ and H+ fluxes of sea urchin sperm. Cell. 1985 Mar;40(3):697–703. doi: 10.1016/0092-8674(85)90218-1. [DOI] [PubMed] [Google Scholar]
  49. Tsien R. W., Hess P., McCleskey E. W., Rosenberg R. L. Calcium channels: mechanisms of selectivity, permeation, and block. Annu Rev Biophys Biophys Chem. 1987;16:265–290. doi: 10.1146/annurev.bb.16.060187.001405. [DOI] [PubMed] [Google Scholar]
  50. Vassilev P. M., Kanazirska M. P., Tien H. T. Ca2+ channels from brain microsomal membranes reconstituted in patch-clamped bilayers. Biochim Biophys Acta. 1987 Feb 26;897(2):324–330. doi: 10.1016/0005-2736(87)90428-7. [DOI] [PubMed] [Google Scholar]
  51. Woll K. H., Leibowitz M. D., Neumcke B., Hille B. A high-conductance anion channel in adult amphibian skeletal muscle. Pflugers Arch. 1987 Dec;410(6):632–640. doi: 10.1007/BF00581324. [DOI] [PubMed] [Google Scholar]
  52. Woll K. H., Neumcke B. Conductance properties and voltage dependence of an anion channel in amphibian skeletal muscle. Pflugers Arch. 1987 Dec;410(6):641–647. doi: 10.1007/BF00581325. [DOI] [PubMed] [Google Scholar]
  53. Yellen G. Permeation in potassium channels: implications for channel structure. Annu Rev Biophys Biophys Chem. 1987;16:227–246. doi: 10.1146/annurev.bb.16.060187.001303. [DOI] [PubMed] [Google Scholar]
  54. Young G. P., Koide S. S., Goldstein M., Young J. D. Isolation and partial characterization of an ion channel protein from human sperm membranes. Arch Biochem Biophys. 1988 May 1;262(2):491–500. doi: 10.1016/0003-9861(88)90400-6. [DOI] [PubMed] [Google Scholar]
  55. Zimmerberg J., Cohen F. S., Finkelstein A. Fusion of phospholipid vesicles with planar phospholipid bilayer membranes. I. Discharge of vesicular contents across the planar membrane. J Gen Physiol. 1980 Mar;75(3):241–250. doi: 10.1085/jgp.75.3.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. von Tscharner V., Prod'hom B., Baggiolini M., Reuter H. Ion channels in human neutrophils activated by a rise in free cytosolic calcium concentration. 1986 Nov 27-Dec 3Nature. 324(6095):369–372. doi: 10.1038/324369a0. [DOI] [PubMed] [Google Scholar]

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