Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Dec 1;103(6):2333–2342. doi: 10.1083/jcb.103.6.2333

The part played by inositol trisphosphate and calcium in the propagation of the fertilization wave in sea urchin eggs

PMCID: PMC2114628  PMID: 3491080

Abstract

Sea urchin egg activation at fertilization is progressive, beginning at the point of sperm entry and moving across the egg with a velocity of 5 microns/s. This activation wave (Kacser, H., 1955, J. Exp. Biol., 32:451-467) has been suggested to be the result of a progressive release of calcium from a store within the egg cytoplasm (Jaffe, L. F., 1983, Dev. Biol., 99:265-276). The progressive release of calcium may be due to the production of inositol trisphosphate (InsP3), a second messenger. We show here that a wave of calcium release crosses the Lytechinus pictus egg; the peak of the wave travels with a velocity of 5 microns/s; microinjection of InsP3 causes the release of calcium within the egg; calcium release (as judged by fertilization envelope elevation) is abolished by prior injection of the calcium chelator EGTA; neomycin, an inhibitor of InsP3 production, does not prevent the release of calcium in response to InsP3 but does abolish the wave of calcium release; the egg cytoplasm rapidly buffers microinjected calcium; the calcium concentration required to cause fertilization membrane elevation when microinjected is very similar to that required to stimulate the production of InsP3 in vitro; and the progressive fertilization membrane elevation seen after microinjection of calcium buffers appears to be due to diffusion of the buffer across the egg cytoplasm rather than to the induction of the activation wave. We conclude that InsP3 diffuses through the egg cytoplasm much more readily than calcium ions and that calcium-stimulated production of InsP3 and InsP3-induced calcium release from an internal store can account for the progressive release of calcium at fertilization.

Full Text

The Full Text of this article is available as a PDF (2.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ALLEN R. D., GRIFFIN J. L. The time sequence of early events in the fertilization of sea urchin eggs. I. The latent period and the cortical reaction. Exp Cell Res. 1958 Aug;15(1):163–173. doi: 10.1016/0014-4827(58)90072-7. [DOI] [PubMed] [Google Scholar]
  2. Allen D. G., Blinks J. R., Prendergast F. G. Aequorin luminescence: relation of light emission to calcium concentration--a calcium-independent component. Science. 1977 Mar 11;195(4282):996–998. doi: 10.1126/science.841325. [DOI] [PubMed] [Google Scholar]
  3. Baker P. F. Transport and metabolism of calcium ions in nerve. Prog Biophys Mol Biol. 1972;24:177–223. doi: 10.1016/0079-6107(72)90007-7. [DOI] [PubMed] [Google Scholar]
  4. Baker P. F., Whitaker M. J. Influence of ATP and calcium on the cortical reaction in sea urchin eggs. Nature. 1978 Nov 30;276(5687):513–515. doi: 10.1038/276513a0. [DOI] [PubMed] [Google Scholar]
  5. Batty I. R., Nahorski S. R., Irvine R. F. Rapid formation of inositol 1,3,4,5-tetrakisphosphate following muscarinic receptor stimulation of rat cerebral cortical slices. Biochem J. 1985 Nov 15;232(1):211–215. doi: 10.1042/bj2320211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Berridge M. J., Irvine R. F. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984 Nov 22;312(5992):315–321. doi: 10.1038/312315a0. [DOI] [PubMed] [Google Scholar]
  7. Busa W. B., Ferguson J. E., Joseph S. K., Williamson J. R., Nuccitelli R. Activation of frog (Xenopus laevis) eggs by inositol trisphosphate. I. Characterization of Ca2+ release from intracellular stores. J Cell Biol. 1985 Aug;101(2):677–682. doi: 10.1083/jcb.101.2.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chambers E. L., Hinkley R. E. Non-propagated cortical reactions induced by the divalent ionophore A23187 in eggs of the sea urchin, Lytechinus variegatus. Exp Cell Res. 1979 Dec;124(2):441–446. doi: 10.1016/0014-4827(79)90221-0. [DOI] [PubMed] [Google Scholar]
  9. Chandler D. E. Exocytosis in vitro: ultrastructure of the isolated sea urchin egg cortex as seen in platinum replicas. J Ultrastruct Res. 1984 Nov;89(2):198–211. doi: 10.1016/s0022-5320(84)80015-5. [DOI] [PubMed] [Google Scholar]
  10. Ciapa B., Whitaker M. Two phases of inositol polyphosphate and diacylglycerol production at fertilisation. FEBS Lett. 1986 Jan 20;195(1-2):347–351. doi: 10.1016/0014-5793(86)80191-0. [DOI] [PubMed] [Google Scholar]
  11. Clapper D. L., Lee H. C. Inositol trisphosphate induces calcium release from nonmitochondrial stores i sea urchin egg homogenates. J Biol Chem. 1985 Nov 15;260(26):13947–13954. [PubMed] [Google Scholar]
  12. Crabb J. H., Jackson R. C. In vitro reconstitution of exocytosis from plasma membrane and isolated secretory vesicles. J Cell Biol. 1985 Dec;101(6):2263–2273. doi: 10.1083/jcb.101.6.2263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eisen A., Kiehart D. P., Wieland S. J., Reynolds G. T. Temporal sequence and spatial distribution of early events of fertilization in single sea urchin eggs. J Cell Biol. 1984 Nov;99(5):1647–1654. doi: 10.1083/jcb.99.5.1647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gilkey J. C., Jaffe L. F., Ridgway E. B., Reynolds G. T. A free calcium wave traverses the activating egg of the medaka, Oryzias latipes. J Cell Biol. 1978 Feb;76(2):448–466. doi: 10.1083/jcb.76.2.448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Griffin H. D., Sykes M., Hawthorne J. N. Effects of neomycin on calcium and polyphosphoinositide metabolism of guinea pig synaptosomes. J Neurochem. 1980 Mar;34(3):750–752. doi: 10.1111/j.1471-4159.1980.tb11209.x. [DOI] [PubMed] [Google Scholar]
  16. Hamaguchi Y., Hiramoto Y. Activation of sea urchin eggs by microinjection of calcium buffers. Exp Cell Res. 1981 Jul;134(1):171–179. doi: 10.1016/0014-4827(81)90474-2. [DOI] [PubMed] [Google Scholar]
  17. Jaffe L. F. Sources of calcium in egg activation: a review and hypothesis. Dev Biol. 1983 Oct;99(2):265–276. doi: 10.1016/0012-1606(83)90276-2. [DOI] [PubMed] [Google Scholar]
  18. Poenie M., Alderton J., Tsien R. Y., Steinhardt R. A. Changes of free calcium levels with stages of the cell division cycle. Nature. 1985 May 9;315(6015):147–149. doi: 10.1038/315147a0. [DOI] [PubMed] [Google Scholar]
  19. Roos A., Boron W. F. Intracellular pH. Physiol Rev. 1981 Apr;61(2):296–434. doi: 10.1152/physrev.1981.61.2.296. [DOI] [PubMed] [Google Scholar]
  20. SHIMOMURA O., JOHNSON F. H., SAIGA Y. Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea. J Cell Comp Physiol. 1962 Jun;59:223–239. doi: 10.1002/jcp.1030590302. [DOI] [PubMed] [Google Scholar]
  21. SUGIYAMA M. Physiological analysis of the cortical response of the sea urchin egg. Exp Cell Res. 1956 Apr;10(2):364–376. doi: 10.1016/0014-4827(56)90009-x. [DOI] [PubMed] [Google Scholar]
  22. Shen S. S., Steinhardt R. A. Direct measurement of intracellular pH during metabolic derepression of the sea urchin egg. Nature. 1978 Mar 16;272(5650):253–254. doi: 10.1038/272253a0. [DOI] [PubMed] [Google Scholar]
  23. Smith G. L., Miller D. J. Potentiometric measurements of stoichiometric and apparent affinity constants of EGTA for protons and divalent ions including calcium. Biochim Biophys Acta. 1985 May 8;839(3):287–299. doi: 10.1016/0304-4165(85)90011-x. [DOI] [PubMed] [Google Scholar]
  24. Steinhardt R., Zucker R., Schatten G. Intracellular calcium release at fertilization in the sea urchin egg. Dev Biol. 1977 Jul 1;58(1):185–196. doi: 10.1016/0012-1606(77)90084-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]
  26. Tsien R. Y., Rink T. J. Neutral carrier ion-selective microelectrodes for measurement of intracellular free calcium. Biochim Biophys Acta. 1980 Jul;599(2):623–638. doi: 10.1016/0005-2736(80)90205-9. [DOI] [PubMed] [Google Scholar]
  27. Turner P. R., Jaffe L. A., Fein A. Regulation of cortical vesicle exocytosis in sea urchin eggs by inositol 1,4,5-trisphosphate and GTP-binding protein. J Cell Biol. 1986 Jan;102(1):70–76. doi: 10.1083/jcb.102.1.70. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Whitaker M. J., Baker P. F. Calcium-dependent exocytosis in an in vitro secretory granule plasma membrane preparation from sea urchin eggs and the effects of some inhibitors of cytoskeletal function. Proc R Soc Lond B Biol Sci. 1983 Jul 22;218(1213):397–413. doi: 10.1098/rspb.1983.0047. [DOI] [PubMed] [Google Scholar]
  29. Whitaker M. J., Steinhardt R. A. Ionic regulation of egg activation. Q Rev Biophys. 1982 Nov;15(4):593–666. doi: 10.1017/s0033583500003760. [DOI] [PubMed] [Google Scholar]
  30. Whitaker M. J., Steinhardt R. A. The relation between the increase in reduced nicotinamide nucleotides and the initiation of DNA synthesis in sea urchin eggs. Cell. 1981 Jul;25(1):95–103. doi: 10.1016/0092-8674(81)90234-8. [DOI] [PubMed] [Google Scholar]
  31. Whitaker M., Aitchison M. Calcium-dependent polyphosphoinositide hydrolysis is associated with exocytosis in vitro. FEBS Lett. 1985 Mar 11;182(1):119–124. doi: 10.1016/0014-5793(85)81167-4. [DOI] [PubMed] [Google Scholar]
  32. Wilson D. B., Connolly T. M., Bross T. E., Majerus P. W., Sherman W. R., Tyler A. N., Rubin L. J., Brown J. E. Isolation and characterization of the inositol cyclic phosphate products of polyphosphoinositide cleavage by phospholipase C. Physiological effects in permeabilized platelets and Limulus photoreceptor cells. J Biol Chem. 1985 Nov 5;260(25):13496–13501. [PubMed] [Google Scholar]
  33. Zucker R. S., Steinhardt R. A. Prevention of the cortical reaction in fertilized sea urchin eggs by injection of calcium-chelating ligands. Biochim Biophys Acta. 1978 Jul 17;541(4):459–466. doi: 10.1016/0304-4165(78)90155-1. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES