Skip to main content
PMC home page
The Journal of General Physiology logoLink to The Journal of General Physiology
. 1974 Dec 1;64(6):643–665. doi: 10.1085/jgp.64.6.643

Changes in Intracellular Free Calcium Concentration during Illumination of Invertebrate Photoreceptors

Detection with Aequorin

J E Brown 1, J R Blinks 1
PMCID: PMC2226181  PMID: 4155426

Abstract

Aequorin, which luminesces in the presence of calcium, was injected into photoreceptor cells of Limulus ventral eye. A bright light stimulus elicited a large increase in aequorin luminescence, the aequorin response, indicating a rise of intracellular calcium ion concentration, Cai. The aequorin response reached a maximum after the peak of the electrical response of the photoreceptor, decayed during a prolonged stimulus, and returned to an undetectable level in the dark. Reduction of Cao reduced the amplitude of the aequorin response by a factor no greater than 3. Raising Cao increased the amplitude of the aequorin response. The aequorin response became smaller when membrane voltage was clamped to successively more positive values. These results indicate that the stimulus-induced rise of Cai may be due in part to a light-induced influx of Ca and in part to release of Ca from an intracellular store. Our findings are consistent with the hypothesis that a rise in Cai is a step in the sequence of events underlying light-adaptation in Limulus ventral photoreceptors. Aequorin was also injected into photoreceptors of Balanus. The aequorin responses were similar to those recorded from Limulus cells in all but two ways: (a) A large sustained aequorin luminescence was measured during a prolonged stimulus, and (b) removal of extracellular calcium reduced the aequorin response to an undetectable level.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Ashley C. C., Ridgway E. B. On the relationships between membrane potential, calcium transient and tension in single barnacle muscle fibres. J Physiol. 1970 Jul;209(1):105–130. doi: 10.1113/jphysiol.1970.sp009158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker P. F., Crawford A. C. Mobility and transport of magnesium in squid giant axons. J Physiol. 1972 Dec;227(3):855–874. doi: 10.1113/jphysiol.1972.sp010062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker P. F., Hodgkin A. L., Ridgway E. B. Depolarization and calcium entry in squid giant axons. J Physiol. 1971 Nov;218(3):709–755. doi: 10.1113/jphysiol.1971.sp009641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beeler G. W., Jr, Reuter H. The relation between membrane potential, membrane currents and activation of contraction in ventricular myocardial fibres. J Physiol. 1970 Mar;207(1):211–229. doi: 10.1113/jphysiol.1970.sp009057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blinks J. R. Calcium transients in striated muscle cells. Eur J Cardiol. 1973 Dec;1(2):135–142. [PubMed] [Google Scholar]
  6. Brown H. M., Hagiwara S., Koike H., Meech R. M. Membrane properties of a barnacle photoreceptor examined by the voltage clamp technique. J Physiol. 1970 Jun;208(2):385–413. doi: 10.1113/jphysiol.1970.sp009127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brown J. E., Lisman J. E. An electrogenic sodium pump in Limulus ventral photoreceptor cells. J Gen Physiol. 1972 Jun;59(6):720–733. doi: 10.1085/jgp.59.6.720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clark A. W., Millecchia R., Mauro A. The ventral photoreceptor cells of Limulus. I. The microanatomy. J Gen Physiol. 1969 Sep;54(3):289–309. doi: 10.1085/jgp.54.3.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ebashi S., Endo M. Calcium ion and muscle contraction. Prog Biophys Mol Biol. 1968;18:123–183. doi: 10.1016/0079-6107(68)90023-0. [DOI] [PubMed] [Google Scholar]
  10. Hagins W. A. The visual process: Excitatory mechanisms in the primary receptor cells. Annu Rev Biophys Bioeng. 1972;1:131–158. doi: 10.1146/annurev.bb.01.060172.001023. [DOI] [PubMed] [Google Scholar]
  11. Hagiwara S., Takahashi K., Junge D. Excitation-contraction coupling in a barnacle muscle fiber as examined with voltage clamp technique. J Gen Physiol. 1968 Feb;51(2):157–175. doi: 10.1085/jgp.51.2.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hastings J. W., Mitchell G., Mattingly P. H., Blinks J. R., Van Leeuwen M. Response of aequorin bioluminescence to rapid changes in calcium concentration. Nature. 1969 Jun 14;222(5198):1047–1050. doi: 10.1038/2221047a0. [DOI] [PubMed] [Google Scholar]
  13. Izutsu K. T., Felton S. P., Siegel I. A., Yoda W. T., Chen A. C. Aequorin: its ionic specificity. Biochem Biophys Res Commun. 1972 Nov 15;49(4):1034–1039. doi: 10.1016/0006-291x(72)90316-6. [DOI] [PubMed] [Google Scholar]
  14. Lisman J. E., Brown J. E. The effects of intracellular iontophoretic injection of calcium and sodium ions on the light response of Limulus ventral photoreceptors. J Gen Physiol. 1972 Jun;59(6):701–719. doi: 10.1085/jgp.59.6.701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lisman J. E., Brown J. E. Two light-induced processes in the photoreceptor cells of Limulus ventral eye. J Gen Physiol. 1971 Nov;58(5):544–561. doi: 10.1085/jgp.58.5.544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Llinás R., Blinks J. R., Nicholson C. Calcium transient in presynaptic terminal of squid giant synapse: detection with aequorin. Science. 1972 Jun 9;176(4039):1127–1129. doi: 10.1126/science.176.4039.1127. [DOI] [PubMed] [Google Scholar]
  17. Millecchia R., Mauro A. The ventral photoreceptor cells of Limulus. 3. A voltage-clamp study. J Gen Physiol. 1969 Sep;54(3):331–351. doi: 10.1085/jgp.54.3.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Millecchia R., Mauro A. The ventral photoreceptor cells of Limulus. II. The basic photoresponse. J Gen Physiol. 1969 Sep;54(3):310–330. doi: 10.1085/jgp.54.3.310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Murray G. C. Intracellular absorption difference spectrum of Limulus extra-ocular photolabile pigment. Science. 1966 Dec 2;154(3753):1182–1183. doi: 10.1126/science.154.3753.1182. [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. SHIMOMURA O., JOHNSON F. H., SAIGA Y. FURTHER DATA ON THE BIOLUMINESCENT PROTEIN, AEQUORIN. J Cell Physiol. 1963 Aug;62:1–8. doi: 10.1002/jcp.1030620102. [DOI] [PubMed] [Google Scholar]
  22. Shimomura O., Johnson F. H. Calcium binding, quantum yield, and emitting molecule in aequorin bioluminescence. Nature. 1970 Sep 26;227(5265):1356–1357. doi: 10.1038/2271356a0. [DOI] [PubMed] [Google Scholar]
  23. Shimomura O., Johnson F. H. Further data on the specificity of aequorin luminescence to calcium. Biochem Biophys Res Commun. 1973 Jul 17;53(2):490–494. doi: 10.1016/0006-291x(73)90688-8. [DOI] [PubMed] [Google Scholar]
  24. Stinnakre J., Tauc L. Calcium influx in active Aplysia neurones detected by injected aequorin. Nat New Biol. 1973 Mar 28;242(117):113–115. doi: 10.1038/newbio242113b0. [DOI] [PubMed] [Google Scholar]
  25. Veloso D., Guynn R. W., Oskarsson M., Veech R. L. The concentrations of free and bound magnesium in rat tissues. Relative constancy of free Mg 2+ concentrations. J Biol Chem. 1973 Jul 10;248(13):4811–4819. [PubMed] [Google Scholar]

Articles from The Journal of General Physiology are provided here courtesy of The Rockefeller University Press

RESOURCES