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. 1978 Feb;275:357–376. doi: 10.1113/jphysiol.1978.sp012194

Changes in the intracellular concentration of free calcium ions in a pace-maker neurone, measured with the metallochromic indicator dye arsenazo III.

A L Gorman, M V Thomas
PMCID: PMC1282549  PMID: 633127

Abstract

1. The bursting pace-maker R-15 cell of Aplysia was injected with the Ca2+ sensitive dye arsenazo III. Changes in absorbance were measured with a differential spectrophotometer to monitor changes in free intracellular Ca2+, [Ca-a], during activity. 2. Dye absorbance increased during each pace-maker-induced burst of action potentials and decreased during the hyperpolarizing phase of the pace-maker cycle. 3. The increase in dye absorbance was, at least in part, dependent upon action potential discharge and was greater when action potential duration was prolonged by treatment with tetraethylammonium chloride. 4. Changes in dye absorbance occurred under voltage clamp conditions when the membrance was depolarized 5-15 mV from a holding potential near the resting potential and were larger with greater step depolarizations. 5. These changes were completely blocked by the addition of 3mM-La3+ to normal ASW. 6. The ratio of the absorbance change between two pairs of wave-lengths during the pace-maker cycle was compared with the ratio observed following injection of Ca2+, Mg2+ and H+ ions. The ratio for the pace-maker cycle was well matched by that for Ca2+ injection, but not by that for injection of Mg2+ or H+. 7. Intracellular Ca2+ injections which increased [Ca]1 to the same amount as occurred during the pace-maker cycle also produced an outward current of sufficient magnitude to account for the post-burst hyperpolarization. 8. Depolarization of the cell membrane by extrinsic current during the burst increased and prolonged the change in dye absorbance as well as the post-burst hyperpolarization. 9 It is suggested that Ca2+ enters during the pace-maker cycle, thereby increasing [Ca]i, and that this increase is sufficient to activate an outward current carried by K+ ions which causes or contributes to the post-burst hyperpolarization.

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Selected References

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  1. ARVANITAKI A., CHALAZONITIS N. Configurations modales de l'activité, propres à différents neurones d'un mëme centre. J Physiol (Paris) 1958 Mar;50(2):122–125. [PubMed] [Google Scholar]
  2. Alving B. O. Spontaneous activity in isolated somata of Aplysia pacemaker naurons. J Gen Physiol. 1968 Jan;51(1):29–45. doi: 10.1085/jgp.51.1.29. [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. 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]
  5. Blaustein M. P. The interrelationship between sodium and calcium fluxes across cell membranes. Rev Physiol Biochem Pharmacol. 1974;70:33–82. doi: 10.1007/BFb0034293. [DOI] [PubMed] [Google Scholar]
  6. Brown J. E., Brown P. K., Pinto L. H. Detection of light-induced changes of intracellular ionized calcium concentration in Limulus ventral photoreceptors using arsenazo III. J Physiol. 1977 May;267(2):299–320. doi: 10.1113/jphysiol.1977.sp011814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brown J. E., Cohen L. B., De Weer P., Pinto L. H., Ross W. N., Salzberg B. M. Rapid changes in intracellular free calcium concentration. Detection by metallochromic indicator dyes in squid giant axon. Biophys J. 1975 Nov;15(11):1155–1160. doi: 10.1016/S0006-3495(75)85891-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Carpenter D., Gunn R. The dependence of pacemaker discharge of Aplysia neurons upon Na+ and Ca++. J Cell Physiol. 1970 Feb;75(1):121–127. doi: 10.1002/jcp.1040750114. [DOI] [PubMed] [Google Scholar]
  9. Connor J. A. Time course separation of two inward currents in molluscan neurons. Brain Res. 1977 Jan 7;119(2):487–492. doi: 10.1016/0006-8993(77)90330-4. [DOI] [PubMed] [Google Scholar]
  10. Dipolo R., Requena J., Brinley F. J., Jr, Mullins L. J., Scarpa A., Tiffert T. Ionized calcium concentrations in squid axons. J Gen Physiol. 1976 Apr;67(4):433–467. doi: 10.1085/jgp.67.4.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Eaton D. C. Potassium ion accumulation near a pace-making cell of Aplysia. J Physiol. 1972 Jul;224(2):421–440. doi: 10.1113/jphysiol.1972.sp009903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eckert R., Lux H. D. A voltage-sensitive persistent calcium conductance in neuronal somata of Helix. J Physiol. 1976 Jan;254(1):129–151. doi: 10.1113/jphysiol.1976.sp011225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eckert R., Tillotson D., Ridgway E. B. Voltage-dependent facilitation of Ca2+ entry in voltage-clamped, aequorin-injected molluscan neurons. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1748–1752. doi: 10.1073/pnas.74.4.1748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Geduldig D., Junge D. Sodium and calcium components of action potentials in the Aplysia giant neurone. J Physiol. 1968 Dec;199(2):347–365. doi: 10.1113/jphysiol.1968.sp008657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gola M. Neurones à ondes-salves des mollusques. Variations cycliques lentes des conductances ioniques. Pflugers Arch. 1974;352(1):17–36. doi: 10.1007/BF01061947. [DOI] [PubMed] [Google Scholar]
  16. HAGIWARA S., SAITO N. Voltage-current relations in nerve cell membrane of Onchidium verruculatum. J Physiol. 1959 Oct;148:161–179. doi: 10.1113/jphysiol.1959.sp006279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hagiwara S. Ca spike. Adv Biophys. 1973;4:71–102. [PubMed] [Google Scholar]
  18. Johnston D. Voltage clamp reveals basis for calcium regulation of bursting pacemaker potentials in Aplysia neurons. Brain Res. 1976 May 7;107(2):418–423. doi: 10.1016/0006-8993(76)90239-0. [DOI] [PubMed] [Google Scholar]
  19. Junge D., Stephens C. L. Cyclic variation of potassium conductance in a burst-generating neurone in Aplysia. J Physiol. 1973 Nov;235(1):155–181. doi: 10.1113/jphysiol.1973.sp010382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kendrick N. C. Purification of arsenazo III, a Ca2+-sensitive dye. Anal Biochem. 1976 Dec;76(2):487–501. doi: 10.1016/0003-2697(76)90342-0. [DOI] [PubMed] [Google Scholar]
  21. Meech R. W. Intracellular calcium injection causes increased potassium conductance in Aplysia nerve cells. Comp Biochem Physiol A Comp Physiol. 1972 Jun 1;42(2):493–499. doi: 10.1016/0300-9629(72)90128-4. [DOI] [PubMed] [Google Scholar]
  22. Meech R. W., Standen N. B. Potassium activation in Helix aspersa neurones under voltage clamp: a component mediated by calcium influx. J Physiol. 1975 Jul;249(2):211–239. doi: 10.1113/jphysiol.1975.sp011012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Meech R. W. The sensitivity of Helix aspersa neurones to injected calcium ions. J Physiol. 1974 Mar;237(2):259–277. doi: 10.1113/jphysiol.1974.sp010481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Meech R. W., Thomas R. C. The effect of calcium injection on the intracellular sodium and pH of snail neurones. J Physiol. 1977 Mar;265(3):867–879. doi: 10.1113/jphysiol.1977.sp011749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smith T. G., Jr, Barker J. L., Gainer H. Requirements for bursting pacemaker potential activity in molluscan neurones. Nature. 1975 Feb 6;253(5491):450–452. doi: 10.1038/253450a0. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Thomas M. V., Gorman A. L. Internal calcium changes in a bursting pacemaker neuron measured with arsenazo III. Science. 1977 Apr 29;196(4289):531–533. doi: 10.1126/science.850795. [DOI] [PubMed] [Google Scholar]
  28. Thomas M. V. Microelectrode amplifier with improved method of input-capacitance neutralisation. Med Biol Eng Comput. 1977 Jul;15(4):450–454. doi: 10.1007/BF02458001. [DOI] [PubMed] [Google Scholar]
  29. Thomas R. C. The effect of carbon dioxide on the intracellular pH and buffering power of snail neurones. J Physiol. 1976 Mar;255(3):715–735. doi: 10.1113/jphysiol.1976.sp011305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wilson W. A., Wachtel H. Negative resistance characteristic essential for the maintenance of slow oscillations in bursting neurons. Science. 1974 Dec 6;186(4167):932–934. doi: 10.1126/science.186.4167.932. [DOI] [PubMed] [Google Scholar]

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