Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Oct 1;135(1):181–190. doi: 10.1083/jcb.135.1.181

Calcium waves along the cleavage furrows in cleavage-stage Xenopus embryos and its inhibition by heparin

PMCID: PMC2121016  PMID: 8858172

Abstract

Calcium signaling is known to be associated with cytokinesis; however, the detailed spatio-temporal pattern of calcium dynamics has remained unclear. We have studied changes of intracellular free calcium in cleavage-stage Xenopus embryos using fluorescent calcium indicator dyes, mainly Calcium Green-1. Cleavage formation was followed by calcium transients that localized to cleavage furrows and propagated along the furrows as calcium waves. The calcium transients at the cleavage furrows were observed at each cleavage furrow at least until blastula stage. The velocity of the calcium waves at the first cleavage furrow was approximately 3 microns/s, which was much slower than that associated with fertilization/egg activation. These calcium waves traveled only along the cleavage furrows and not in the direction orthogonal to the furrows. These observations imply that there exists an intracellular calcium-releasing activity specifically associated with cleavage furrows. The calcium waves occurred in the absence of extracellular calcium and were inhibited in embryos injected with heparin an inositol 1,4,5-trisphosphate (InsP3) receptor antagonist. These results suggest that InsP3 receptor-mediated calcium mobilization plays an essential role in calcium wave formation at the cleavage furrows.

Full Text

The Full Text of this article is available as a PDF (6.3 MB).

Selected References

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

  1. Bi G. Q., Alderton J. M., Steinhardt R. A. Calcium-regulated exocytosis is required for cell membrane resealing. J Cell Biol. 1995 Dec;131(6 Pt 2):1747–1758. doi: 10.1083/jcb.131.6.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Busa W. B., Nuccitelli R. An elevated free cytosolic Ca2+ wave follows fertilization in eggs of the frog, Xenopus laevis. J Cell Biol. 1985 Apr;100(4):1325–1329. doi: 10.1083/jcb.100.4.1325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Byers T. J., Armstrong P. B. Membrane protein redistribution during Xenopus first cleavage. J Cell Biol. 1986 Jun;102(6):2176–2184. doi: 10.1083/jcb.102.6.2176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cao L. G., Wang Y. L. Mechanism of the formation of contractile ring in dividing cultured animal cells. II. Cortical movement of microinjected actin filaments. J Cell Biol. 1990 Nov;111(5 Pt 1):1905–1911. doi: 10.1083/jcb.111.5.1905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang D. C., Meng C. A localized elevation of cytosolic free calcium is associated with cytokinesis in the zebrafish embryo. J Cell Biol. 1995 Dec;131(6 Pt 1):1539–1545. doi: 10.1083/jcb.131.6.1539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fluck R. A., Miller A. L., Jaffe L. F. Slow calcium waves accompany cytokinesis in medaka fish eggs. J Cell Biol. 1991 Dec;115(5):1259–1265. doi: 10.1083/jcb.115.5.1259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Furuichi T., Kohda K., Miyawaki A., Mikoshiba K. Intracellular channels. Curr Opin Neurobiol. 1994 Jun;4(3):294–303. doi: 10.1016/0959-4388(94)90089-2. [DOI] [PubMed] [Google Scholar]
  8. Grandin N., Charbonneau M. Intracellular free calcium oscillates during cell division of Xenopus embryos. J Cell Biol. 1991 Feb;112(4):711–718. doi: 10.1083/jcb.112.4.711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Han J. K., Fukami K., Nuccitelli R. Reducing inositol lipid hydrolysis, Ins(1,4,5)P3 receptor availability, or Ca2+ gradients lengthens the duration of the cell cycle in Xenopus laevis blastomeres. J Cell Biol. 1992 Jan;116(1):147–156. doi: 10.1083/jcb.116.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hara K., Tydeman P., Kirschner M. A cytoplasmic clock with the same period as the division cycle in Xenopus eggs. Proc Natl Acad Sci U S A. 1980 Jan;77(1):462–466. doi: 10.1073/pnas.77.1.462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hepler P. K. The role of calcium in cell division. Cell Calcium. 1994 Oct;16(4):322–330. doi: 10.1016/0143-4160(94)90096-5. [DOI] [PubMed] [Google Scholar]
  12. Keating T. J., Cork R. J., Robinson K. R. Intracellular free calcium oscillations in normal and cleavage-blocked embryos and artificially activated eggs of Xenopus laevis. J Cell Sci. 1994 Aug;107(Pt 8):2229–2237. doi: 10.1242/jcs.107.8.2229. [DOI] [PubMed] [Google Scholar]
  13. Kubota H. Y., Yoshimoto Y., Hiramoto Y. Oscillation of intracellular free calcium in cleaving and cleavage-arrested embryos of Xenopus laevis. Dev Biol. 1993 Dec;160(2):512–518. doi: 10.1006/dbio.1993.1325. [DOI] [PubMed] [Google Scholar]
  14. Kume S., Muto A., Aruga J., Nakagawa T., Michikawa T., Furuichi T., Nakade S., Okano H., Mikoshiba K. The Xenopus IP3 receptor: structure, function, and localization in oocytes and eggs. Cell. 1993 May 7;73(3):555–570. doi: 10.1016/0092-8674(93)90142-d. [DOI] [PubMed] [Google Scholar]
  15. McDougall A., Sardet C. Function and characteristics of repetitive calcium waves associated with meiosis. Curr Biol. 1995 Mar 1;5(3):318–328. doi: 10.1016/s0960-9822(95)00062-5. [DOI] [PubMed] [Google Scholar]
  16. Mittal B., Sanger J. M., Sanger J. W. Visualization of myosin in living cells. J Cell Biol. 1987 Oct;105(4):1753–1760. doi: 10.1083/jcb.105.4.1753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nuccitelli R., Yim D. L., Smart T. The sperm-induced Ca2+ wave following fertilization of the Xenopus egg requires the production of Ins(1, 4, 5)P3. Dev Biol. 1993 Jul;158(1):200–212. doi: 10.1006/dbio.1993.1179. [DOI] [PubMed] [Google Scholar]
  18. Parys J. B., Sernett S. W., DeLisle S., Snyder P. M., Welsh M. J., Campbell K. P. Isolation, characterization, and localization of the inositol 1,4,5-trisphosphate receptor protein in Xenopus laevis oocytes. J Biol Chem. 1992 Sep 15;267(26):18776–18782. [PubMed] [Google Scholar]
  19. 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]
  20. Snow P., Nuccitelli R. Calcium buffer injections delay cleavage in Xenopus laevis blastomeres. J Cell Biol. 1993 Jul;122(2):387–394. doi: 10.1083/jcb.122.2.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stricker S. A. Time-lapse confocal imaging of calcium dynamics in starfish embryos. Dev Biol. 1995 Aug;170(2):496–518. doi: 10.1006/dbio.1995.1232. [DOI] [PubMed] [Google Scholar]
  22. Sugiyama T., Furuya A., Monkawa T., Yamamoto-Hino M., Satoh S., Ohmori K., Miyawaki A., Hanai N., Mikoshiba K., Hasegawa M. Monoclonal antibodies distinctively recognizing the subtypes of inositol 1,4,5-trisphosphate receptor: application to the studies on inflammatory cells. FEBS Lett. 1994 Nov 7;354(2):149–154. doi: 10.1016/0014-5793(94)01099-4. [DOI] [PubMed] [Google Scholar]
  23. Sullivan K. M., Busa W. B., Wilson K. L. Calcium mobilization is required for nuclear vesicle fusion in vitro: implications for membrane traffic and IP3 receptor function. Cell. 1993 Jul 2;73(7):1411–1422. doi: 10.1016/0092-8674(93)90366-x. [DOI] [PubMed] [Google Scholar]
  24. Tombes R. M., Simerly C., Borisy G. G., Schatten G. Meiosis, egg activation, and nuclear envelope breakdown are differentially reliant on Ca2+, whereas germinal vesicle breakdown is Ca2+ independent in the mouse oocyte. J Cell Biol. 1992 May;117(4):799–811. doi: 10.1083/jcb.117.4.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tsien R., Pozzan T. Measurement of cytosolic free Ca2+ with quin2. Methods Enzymol. 1989;172:230–262. doi: 10.1016/s0076-6879(89)72017-6. [DOI] [PubMed] [Google Scholar]
  26. Yamakita Y., Yamashiro S., Matsumura F. In vivo phosphorylation of regulatory light chain of myosin II during mitosis of cultured cells. J Cell Biol. 1994 Jan;124(1-2):129–137. doi: 10.1083/jcb.124.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES