Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Apr;87(7):2813–2817. doi: 10.1073/pnas.87.7.2813

Calcium wave evoked by activation of endogenous or exogenously expressed receptors in Xenopus oocytes.

G Brooker 1, T Seki 1, D Croll 1, C Wahlestedt 1
PMCID: PMC53781  PMID: 2157216

Abstract

The mRNA encoding the cloned substance K receptor was microinjected into Xenopus laevis oocytes. After expression of the mRNA, Ca2+ was imaged in the oocytes with a digital imaging fluorescence microscopy system using the Ca2(+)-sensitive dyes fura-2 and fluo-3. Application of substance K caused a dose-related wave of Ca2+ mobilization to spread from a focus and to elevate the Ca2+ concentration in the oocyte. Activation of endogenous muscarinic or angiotensin II receptors in noninjected oocytes evoked a similar response. The Ca2+ rise in oocytes induced by substance K was due to internal Ca2+ mobilization and was independent of external Ca2+, since it occurred in Ca2(+)-free medium fortified with 2 mM EGTA. The Ca2+ imaging was well correlated with ion current measurements of voltage-clamped oocytes. Imaging, in addition to detecting the spatial spread of Ca2+ across the cell, was at least as sensitive as voltage clamping and much faster when screening oocytes for the expression of receptor mRNAs that stimulate Ca2+ mobilization. While it is known that fertilization of Xenopus eggs causes a spreading wave of Ca2+ mobilization, we found that activation of either native or newly expressed receptors in oocytes causes a similar change in Ca2+ distribution.

Full text

PDF
2813

Images in this article

2814Image
on p.2814
2815Image
on p.2815
2816Image
on p.2816

Selected References

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

  1. Ambler S. K., Poenie M., Tsien R. Y., Taylor P. Agonist-stimulated oscillations and cycling of intracellular free calcium in individual cultured muscle cells. J Biol Chem. 1988 Feb 5;263(4):1952–1959. [PubMed] [Google Scholar]
  2. Berridge M. J., Galione A. Cytosolic calcium oscillators. FASEB J. 1988 Dec;2(15):3074–3082. doi: 10.1096/fasebj.2.15.2847949. [DOI] [PubMed] [Google Scholar]
  3. Berridge M. J. Inositol trisphosphate-induced membrane potential oscillations in Xenopus oocytes. J Physiol. 1988 Sep;403:589–599. doi: 10.1113/jphysiol.1988.sp017266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Connor J. A. Digital imaging of free calcium changes and of spatial gradients in growing processes in single, mammalian central nervous system cells. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6179–6183. doi: 10.1073/pnas.83.16.6179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cork R. J., Cicirelli M. F., Robinson K. R. A rise in cytosolic calcium is not necessary for maturation of Xenopus laevis oocytes. Dev Biol. 1987 May;121(1):41–47. doi: 10.1016/0012-1606(87)90136-9. [DOI] [PubMed] [Google Scholar]
  6. Dascal N. The use of Xenopus oocytes for the study of ion channels. CRC Crit Rev Biochem. 1987;22(4):317–387. doi: 10.3109/10409238709086960. [DOI] [PubMed] [Google Scholar]
  7. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  8. Julius D., MacDermott A. B., Axel R., Jessell T. M. Molecular characterization of a functional cDNA encoding the serotonin 1c receptor. Science. 1988 Jul 29;241(4865):558–564. doi: 10.1126/science.3399891. [DOI] [PubMed] [Google Scholar]
  9. Kline D., Simoncini L., Mandel G., Maue R. A., Kado R. T., Jaffe L. A. Fertilization events induced by neurotransmitters after injection of mRNA in Xenopus eggs. Science. 1988 Jul 22;241(4864):464–467. doi: 10.1126/science.3134693. [DOI] [PubMed] [Google Scholar]
  10. Lupu-Meiri M., Shapira H., Oron Y. Hemispheric asymmetry of rapid chloride responses to inositol trisphosphate and calcium in Xenopus oocytes. FEBS Lett. 1988 Nov 21;240(1-2):83–87. doi: 10.1016/0014-5793(88)80344-2. [DOI] [PubMed] [Google Scholar]
  11. Masu Y., Nakayama K., Tamaki H., Harada Y., Kuno M., Nakanishi S. cDNA cloning of bovine substance-K receptor through oocyte expression system. 1987 Oct 29-Nov 4Nature. 329(6142):836–838. doi: 10.1038/329836a0. [DOI] [PubMed] [Google Scholar]
  12. Minta A., Kao J. P., Tsien R. Y. Fluorescent indicators for cytosolic calcium based on rhodamine and fluorescein chromophores. J Biol Chem. 1989 May 15;264(14):8171–8178. [PubMed] [Google Scholar]
  13. Sandberg K., Markwick A. J., Trinh D. P., Catt K. J. Calcium mobilization by angiotensin II and neurotransmitter receptors expressed in Xenopus laevis oocytes. FEBS Lett. 1988 Dec 5;241(1-2):177–180. doi: 10.1016/0014-5793(88)81055-x. [DOI] [PubMed] [Google Scholar]
  14. Smith A. A., Brooker T., Brooker G. Expression of rat mRNA coding for hormone-stimulated adenylate cyclase in Xenopus oocytes. FASEB J. 1987 Nov;1(5):380–387. doi: 10.1096/fasebj.1.5.2824269. [DOI] [PubMed] [Google Scholar]
  15. Soreq H. The biosynthesis of biologically active proteins in mRNA-microinjected Xenopus oocytes. CRC Crit Rev Biochem. 1985;18(3):199–238. doi: 10.3109/10409238509085134. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES