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. 1992 Oct;63(4):1146–1164. doi: 10.1016/S0006-3495(92)81670-3

Evaluation of cellular mechanisms for modulation of calcium transients using a mathematical model of fura-2 Ca2+ imaging in Aplysia sensory neurons.

H Blumenfeld 1, L Zablow 1, B Sabatini 1
PMCID: PMC1262252  PMID: 1420931

Abstract

A theoretical model of [Ca++]i diffusion, buffering, and extrusion was developed for Aplysia sensory neurons, and integrated with the measured optical transfer function of our fura-2 microscopic recording system, in order to fully simulate fura-2 video or photomultiplier tube measurements of [Ca++]i. This allowed an analysis of the spatial and temporal distortions introduced during each step of fura-2 measurements of [Ca++]i in cells. In addition, the model was used to evaluate the plausibility of several possible mechanisms for modulating [Ca++]i transients evoked by action potentials. The results of the model support prior experimental work (Blumenfeld, Spira, Kandel, and Siegelbaum, 1990. Neuron. 5: 487-499), suggesting that 5-HT and FMRFamide modulate action potential-induced [Ca++]i transients in Aplysia sensory neurons through changes in Ca++ influx, and not through changes in [Ca++]i homeostasis or release from internal stores.

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Selected References

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  1. Abrams T. W., Castellucci V. F., Camardo J. S., Kandel E. R., Lloyd P. E. Two endogenous neuropeptides modulate the gill and siphon withdrawal reflex in Aplysia by presynaptic facilitation involving cAMP-dependent closure of a serotonin-sensitive potassium channel. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7956–7960. doi: 10.1073/pnas.81.24.7956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adler E. M., Augustine G. J., Duffy S. N., Charlton M. P. Alien intracellular calcium chelators attenuate neurotransmitter release at the squid giant synapse. J Neurosci. 1991 Jun;11(6):1496–1507. doi: 10.1523/JNEUROSCI.11-06-01496.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ahmed Z., Connor J. A. Calcium regulation by and buffer capacity of molluscan neurons during calcium transients. Cell Calcium. 1988 Apr;9(2):57–69. doi: 10.1016/0143-4160(88)90025-5. [DOI] [PubMed] [Google Scholar]
  4. Andresen M. C., Brown A. M., Yasui S. The role of diffusion in the photoresponse of an extraretinal photoreceptor of Aplysia. J Physiol. 1979 Feb;287:283–301. doi: 10.1113/jphysiol.1979.sp012659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Anglister L., Farber I. C., Shahar A., Grinvald A. Localization of voltage-sensitive calcium channels along developing neurites: their possible role in regulating neurite elongation. Dev Biol. 1982 Dec;94(2):351–365. doi: 10.1016/0012-1606(82)90353-0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Barish M. E., Thompson S. H. Calcium buffering and slow recovery kinetics of calcium-dependent outward current in molluscan neurones. J Physiol. 1983 Apr;337:201–219. doi: 10.1113/jphysiol.1983.sp014620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Belardetti F., Kandel E. R., Siegelbaum S. A. Neuronal inhibition by the peptide FMRFamide involves opening of S K+ channels. Nature. 1987 Jan 8;325(7000):153–156. doi: 10.1038/325153a0. [DOI] [PubMed] [Google Scholar]
  9. Blaustein M. P. Calcium transport and buffering in neurons. Trends Neurosci. 1988 Oct;11(10):438–443. doi: 10.1016/0166-2236(88)90195-6. [DOI] [PubMed] [Google Scholar]
  10. Blaustein M. P., Hodgkin A. L. The effect of cyanide on the efflux of calcium from squid axons. J Physiol. 1969 Feb;200(2):497–527. doi: 10.1113/jphysiol.1969.sp008704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Blaustein M. P., Ratzlaff R. W., Schweitzer E. S. Calcium buffering in presynaptic nerve terminals. II. Kinetic properties of the nonmitochondrial Ca sequestration mechanism. J Gen Physiol. 1978 Jul;72(1):43–66. doi: 10.1085/jgp.72.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Blumenfeld H., Spira M. E., Kandel E. R., Siegelbaum S. A. Facilitatory and inhibitory transmitters modulate calcium influx during action potentials in aplysia sensory neurons. Neuron. 1990 Oct;5(4):487–499. doi: 10.1016/0896-6273(90)90088-w. [DOI] [PubMed] [Google Scholar]
  13. Boyle M. B., Klein M., Smith S. J., Kandel E. R. Serotonin increases intracellular Ca2+ transients in voltage-clamped sensory neurons of Aplysia californica. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7642–7646. doi: 10.1073/pnas.81.23.7642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Brezina V., Eckert R., Erxleben C. Modulation of potassium conductances by an endogenous neuropeptide in neurones of Aplysia californica. J Physiol. 1987 Jan;382:267–290. doi: 10.1113/jphysiol.1987.sp016367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Carafoli E. Intracellular calcium homeostasis. Annu Rev Biochem. 1987;56:395–433. doi: 10.1146/annurev.bi.56.070187.002143. [DOI] [PubMed] [Google Scholar]
  16. Chad J. E., Eckert R. Calcium domains associated with individual channels can account for anomalous voltage relations of CA-dependent responses. Biophys J. 1984 May;45(5):993–999. doi: 10.1016/S0006-3495(84)84244-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Connor J. A., Kretz R., Shapiro E. Calcium levels measured in a presynaptic neurone of Aplysia under conditions that modulate transmitter release. J Physiol. 1986 Jun;375:625–642. doi: 10.1113/jphysiol.1986.sp016137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. DiPolo R., Beaugé L. The calcium pump and sodium-calcium exchange in squid axons. Annu Rev Physiol. 1983;45:313–324. doi: 10.1146/annurev.ph.45.030183.001525. [DOI] [PubMed] [Google Scholar]
  19. Dissing S., Nauntofte B., Sten-Knudsen O. Spatial distribution of intracellular, free Ca2+ in isolated rat parotid acini. Pflugers Arch. 1990 Sep;417(1):1–12. doi: 10.1007/BF00370762. [DOI] [PubMed] [Google Scholar]
  20. Edmonds B., Klein M., Dale N., Kandel E. R. Contributions of two types of calcium channels to synaptic transmission and plasticity. Science. 1990 Nov 23;250(4984):1142–1147. doi: 10.1126/science.2174573. [DOI] [PubMed] [Google Scholar]
  21. Fay F. S., Carrington W., Fogarty K. E. Three-dimensional molecular distribution in single cells analysed using the digital imaging microscope. J Microsc. 1989 Feb;153(Pt 2):133–149. [PubMed] [Google Scholar]
  22. Fischmeister R., Horackova M. Variation of intracellular Ca2+ following Ca2+ current in heart. A theoretical study of ionic diffusion inside a cylindrical cell. Biophys J. 1983 Mar;41(3):341–348. doi: 10.1016/S0006-3495(83)84445-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gamble E., Koch C. The dynamics of free calcium in dendritic spines in response to repetitive synaptic input. Science. 1987 Jun 5;236(4806):1311–1315. doi: 10.1126/science.3495885. [DOI] [PubMed] [Google Scholar]
  24. Gorman A. L., Thomas M. V. Intracellular calcium accumulation during depolarization in a molluscan neurone. J Physiol. 1980 Nov;308:259–285. doi: 10.1113/jphysiol.1980.sp013471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Graubard K. Voltage attenuation within Aplysia neurons: the effect of branching pattern. Brain Res. 1975 May 2;88(2):325–332. doi: 10.1016/0006-8993(75)90394-7. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Hernández-Cruz A., Sala F., Adams P. R. Subcellular calcium transients visualized by confocal microscopy in a voltage-clamped vertebrate neuron. Science. 1990 Feb 16;247(4944):858–862. doi: 10.1126/science.2154851. [DOI] [PubMed] [Google Scholar]
  28. Hiraoka Y., Sedat J. W., Agard D. A. Determination of three-dimensional imaging properties of a light microscope system. Partial confocal behavior in epifluorescence microscopy. Biophys J. 1990 Feb;57(2):325–333. doi: 10.1016/S0006-3495(90)82534-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hochner B., Klein M., Schacher S., Kandel E. R. Additional component in the cellular mechanism of presynaptic facilitation contributes to behavioral dishabituation in Aplysia. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8794–8798. doi: 10.1073/pnas.83.22.8794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Hockberger P. E., Tseng H. Y., Connor J. A. Fura-2 measurements of cultured rat Purkinje neurons show dendritic localization of Ca2+ influx. J Neurosci. 1989 Jul;9(7):2272–2284. doi: 10.1523/JNEUROSCI.09-07-02272.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Hollý M., Poledna J. Model of calcium diffusion, binding and membrane transport in the sarcomere of frog skeletal muscle. Gen Physiol Biophys. 1989 Dec;8(6):539–553. [PubMed] [Google Scholar]
  32. Holmes W. R., Levy W. B. Insights into associative long-term potentiation from computational models of NMDA receptor-mediated calcium influx and intracellular calcium concentration changes. J Neurophysiol. 1990 May;63(5):1148–1168. doi: 10.1152/jn.1990.63.5.1148. [DOI] [PubMed] [Google Scholar]
  33. Jackson A. P., Timmerman M. P., Bagshaw C. R., Ashley C. C. The kinetics of calcium binding to fura-2 and indo-1. FEBS Lett. 1987 May 25;216(1):35–39. doi: 10.1016/0014-5793(87)80752-4. [DOI] [PubMed] [Google Scholar]
  34. Kandel E. R., Schwartz J. H. Molecular biology of learning: modulation of transmitter release. Science. 1982 Oct 29;218(4571):433–443. doi: 10.1126/science.6289442. [DOI] [PubMed] [Google Scholar]
  35. Klein M., Kandel E. R. Presynaptic modulation of voltage-dependent Ca2+ current: mechanism for behavioral sensitization in Aplysia californica. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3512–3516. doi: 10.1073/pnas.75.7.3512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Konishi M., Olson A., Hollingworth S., Baylor S. M. Myoplasmic binding of fura-2 investigated by steady-state fluorescence and absorbance measurements. Biophys J. 1988 Dec;54(6):1089–1104. doi: 10.1016/S0006-3495(88)83045-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Lipscombe D., Madison D. V., Poenie M., Reuter H., Tsien R. W., Tsien R. Y. Imaging of cytosolic Ca2+ transients arising from Ca2+ stores and Ca2+ channels in sympathetic neurons. Neuron. 1988 Jul;1(5):355–365. doi: 10.1016/0896-6273(88)90185-7. [DOI] [PubMed] [Google Scholar]
  38. Lipscombe D., Madison D. V., Poenie M., Reuter H., Tsien R. Y., Tsien R. W. Spatial distribution of calcium channels and cytosolic calcium transients in growth cones and cell bodies of sympathetic neurons. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2398–2402. doi: 10.1073/pnas.85.7.2398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Mirolli M., Talbott S. R. The geometrical factors determining the electrotonic properties of a molluscan neurone. J Physiol. 1972 Dec;227(1):19–34. doi: 10.1113/jphysiol.1972.sp010017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Müller W., Connor J. A. Dendritic spines as individual neuronal compartments for synaptic Ca2+ responses. Nature. 1991 Nov 7;354(6348):73–76. doi: 10.1038/354073a0. [DOI] [PubMed] [Google Scholar]
  41. Parnas H., Hovav G., Parnas I. Effect of Ca2+ diffusion on the time course of neurotransmitter release. Biophys J. 1989 May;55(5):859–874. doi: 10.1016/S0006-3495(89)82885-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Requena J., Mullins L. J. Calcium movement in nerve fibres. Q Rev Biophys. 1979 Aug;12(3):371–460. doi: 10.1017/s0033583500005473. [DOI] [PubMed] [Google Scholar]
  43. Sala F., Hernández-Cruz A. Calcium diffusion modeling in a spherical neuron. Relevance of buffering properties. Biophys J. 1990 Feb;57(2):313–324. doi: 10.1016/S0006-3495(90)82533-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Schatzmann H. J. The calcium pump of the surface membrane and of the sarcoplasmic reticulum. Annu Rev Physiol. 1989;51:473–485. doi: 10.1146/annurev.ph.51.030189.002353. [DOI] [PubMed] [Google Scholar]
  45. Siegelbaum S. A., Camardo J. S., Kandel E. R. Serotonin and cyclic AMP close single K+ channels in Aplysia sensory neurones. Nature. 1982 Sep 30;299(5882):413–417. doi: 10.1038/299413a0. [DOI] [PubMed] [Google Scholar]
  46. Silver R. A., Lamb A. G., Bolsover S. R. Calcium hotspots caused by L-channel clustering promote morphological changes in neuronal growth cones. Nature. 1990 Feb 22;343(6260):751–754. doi: 10.1038/343751a0. [DOI] [PubMed] [Google Scholar]
  47. Simon S. M., Llinás R. R. Compartmentalization of the submembrane calcium activity during calcium influx and its significance in transmitter release. Biophys J. 1985 Sep;48(3):485–498. doi: 10.1016/S0006-3495(85)83804-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Smith S. J., Augustine G. J. Calcium ions, active zones and synaptic transmitter release. Trends Neurosci. 1988 Oct;11(10):458–464. doi: 10.1016/0166-2236(88)90199-3. [DOI] [PubMed] [Google Scholar]
  49. Stockbridge N., Moore J. W. Dynamics of intracellular calcium and its possible relationship to phasic transmitter release and facilitation at the frog neuromuscular junction. J Neurosci. 1984 Mar;4(3):803–811. doi: 10.1523/JNEUROSCI.04-03-00803.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Strautman A. F., Cork R. J., Robinson K. R. The distribution of free calcium in transected spinal axons and its modulation by applied electrical fields. J Neurosci. 1990 Nov;10(11):3564–3575. doi: 10.1523/JNEUROSCI.10-11-03564.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Thayer S. A., Hirning L. D., Miller R. J. The role of caffeine-sensitive calcium stores in the regulation of the intracellular free calcium concentration in rat sympathetic neurons in vitro. Mol Pharmacol. 1988 Nov;34(5):664–673. [PubMed] [Google Scholar]
  52. Thayer S. A., Miller R. J. Regulation of the intracellular free calcium concentration in single rat dorsal root ganglion neurones in vitro. J Physiol. 1990 Jun;425:85–115. doi: 10.1113/jphysiol.1990.sp018094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Thayer S. A., Perney T. M., Miller R. J. Regulation of calcium homeostasis in sensory neurons by bradykinin. J Neurosci. 1988 Nov;8(11):4089–4097. doi: 10.1523/JNEUROSCI.08-11-04089.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Thompson S., Coombs J. Spatial distribution of Ca currents in molluscan neuron cell bodies and regional differences in the strength of inactivation. J Neurosci. 1988 Jun;8(6):1929–1939. doi: 10.1523/JNEUROSCI.08-06-01929.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Tillotson D., Gorman A. L. Non-uniform Ca2+ buffer distribution in a nerve cell body. Nature. 1980 Aug 21;286(5775):816–817. doi: 10.1038/286816a0. [DOI] [PubMed] [Google Scholar]
  56. Timmerman M. P., Ashley C. C. Fura-2 diffusion and its use as an indicator of transient free calcium changes in single striated muscle cells. FEBS Lett. 1986 Dec 1;209(1):1–8. doi: 10.1016/0014-5793(86)81073-0. [DOI] [PubMed] [Google Scholar]
  57. Williams D. A., Fogarty K. E., Tsien R. Y., Fay F. S. Calcium gradients in single smooth muscle cells revealed by the digital imaging microscope using Fura-2. Nature. 1985 Dec 12;318(6046):558–561. doi: 10.1038/318558a0. [DOI] [PubMed] [Google Scholar]
  58. Zucker R. S., Fogelson A. L. Relationship between transmitter release and presynaptic calcium influx when calcium enters through discrete channels. Proc Natl Acad Sci U S A. 1986 May;83(9):3032–3036. doi: 10.1073/pnas.83.9.3032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Zucker R. S., Stockbridge N. Presynaptic calcium diffusion and the time courses of transmitter release and synaptic facilitation at the squid giant synapse. J Neurosci. 1983 Jun;3(6):1263–1269. doi: 10.1523/JNEUROSCI.03-06-01263.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

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