Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Feb 2;124(4):463–474. doi: 10.1083/jcb.124.4.463

Intracellular calcium and cAMP regulate directional pigment movements in teleost erythrophores

PMCID: PMC2119921  PMID: 8106546

Abstract

Teleost pigment cells (erythrophores and melanophores) are useful models for studying the regulation of rapid, microtubule-dependent organelle transport. Previous studies suggest that melanophores regulate the direction of pigment movements via changes in intracellular cAMP (Rozdzial and Haimo, 1986a; Sammak et al., 1992), whereas erythrophores may use calcium- (Ca(2+)-) based regulation (Luby- Phelps and Porter, 1982; McNiven and Ward, 1988). Despite these observations, there have been no direct measurements in intact erythrophores or any cell type correlating changes of intracellular free Ca2+ ([Ca2+]i) with organelle movements. Here we demonstrate that extracellular Ca2+ is necessary and that a Ca2+ influx via microinjection is sufficient to induce pigment aggregation in erythrophores, but not melanophores of squirrel fish. Using the Ca(2+)- sensitive indicator, Fura-2, we demonstrate that [Ca2+]i rises dramatically concomitant with aggregation of pigment granules in erythrophores, but not melanophores. In addition, we find that an erythrophore stimulated to aggregate pigment will immediately transmit a rise in [Ca2+]i to neighboring cells, suggesting that these cells are electrically coupled. Surprisingly, we find that a fall in [Ca2+]i is not sufficient to induce pigment dispersion in erythrophores, contrary to the findings obtained with the ionophore and lysed-cell models (Luby- Phelps and Porter, 1982; McNiven and Ward, 1988). We find that a rise in intracellular cAMP ([cAMP]i) induces pigment dispersion, and that this dispersive stimulus can be overridden by an aggregation stimulus, suggesting that both high [cAMP]i and low [Ca2+]i are necessary to produce pigment dispersion in erythrophores.

Full Text

The Full Text of this article is available as a PDF (6.5 MB).

Selected References

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

  1. Barylko B., Wagner M. C., Reizes O., Albanesi J. P. Purification and characterization of a mammalian myosin I. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):490–494. doi: 10.1073/pnas.89.2.490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beckerle M. C., Porter K. R. Inhibitors of dynein activity block intracellular transport in erythrophores. Nature. 1982 Feb 25;295(5851):701–703. doi: 10.1038/295701a0. [DOI] [PubMed] [Google Scholar]
  3. Byers H. R., Porter K. R. Transformations in the structure of the cytoplasmic ground substance in erythrophores during pigment aggregation and dispersion. I. A study using whole-cell preparations in stereo high voltage electron microscopy. J Cell Biol. 1977 Nov;75(2 Pt 1):541–558. doi: 10.1083/jcb.75.2.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cheney R. E., Mooseker M. S. Unconventional myosins. Curr Opin Cell Biol. 1992 Feb;4(1):27–35. doi: 10.1016/0955-0674(92)90055-h. [DOI] [PubMed] [Google Scholar]
  5. Clark T. G., Rosenbaum J. L. Pigment particle translocation in detergent-permeabilized melanophores of Fundulus heteroclitus. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4655–4659. doi: 10.1073/pnas.79.15.4655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Collins K., Sellers J. R., Matsudaira P. Calmodulin dissociation regulates brush border myosin I (110-kD-calmodulin) mechanochemical activity in vitro. J Cell Biol. 1990 Apr;110(4):1137–1147. doi: 10.1083/jcb.110.4.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Conzelman K. A., Mooseker M. S. The 110-kD protein-calmodulin complex of the intestinal microvillus is an actin-activated MgATPase. J Cell Biol. 1987 Jul;105(1):313–324. doi: 10.1083/jcb.105.1.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Exton J. H. Mechanisms involved in alpha-adrenergic phenomena. Am J Physiol. 1985 Jun;248(6 Pt 1):E633–E647. doi: 10.1152/ajpendo.1985.248.6.E633. [DOI] [PubMed] [Google Scholar]
  9. Gibbons B. H., Gibbons I. R. Calcium-induced quiescence in reactivated sea urchin sperm. J Cell Biol. 1980 Jan;84(1):13–27. doi: 10.1083/jcb.84.1.13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Haimo L. T., Rozdzial M. M. Lysed chromatophores: a model system for the study of bidirectional organelle transport. Methods Cell Biol. 1989;31:3–24. doi: 10.1016/s0091-679x(08)61599-x. [DOI] [PubMed] [Google Scholar]
  12. Hollenbeck P. J. Phosphorylation of neuronal kinesin heavy and light chains in vivo. J Neurochem. 1993 Jun;60(6):2265–2275. doi: 10.1111/j.1471-4159.1993.tb03513.x. [DOI] [PubMed] [Google Scholar]
  13. Hollenbeck P. J. Products of endocytosis and autophagy are retrieved from axons by regulated retrograde organelle transport. J Cell Biol. 1993 Apr;121(2):305–315. doi: 10.1083/jcb.121.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Holwill M. E., McGregor J. L. Effects of calcium on flagellar movement in the trypanosome Crithidia oncopelti. J Exp Biol. 1976 Aug;65(1):229–242. doi: 10.1242/jeb.65.1.229. [DOI] [PubMed] [Google Scholar]
  15. Hyams J. S., Borisy G. G. Isolated flagellar apparatus of Chlamydomonas: characterization of forward swimming and alteration of waveform and reversal of motion by calcium ions in vitro. J Cell Sci. 1978 Oct;33:235–253. doi: 10.1242/jcs.33.1.235. [DOI] [PubMed] [Google Scholar]
  16. Lohmann S. M., DeCamilli P., Einig I., Walter U. High-affinity binding of the regulatory subunit (RII) of cAMP-dependent protein kinase to microtubule-associated and other cellular proteins. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6723–6727. doi: 10.1073/pnas.81.21.6723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Luby-Phelps K., Porter K. R. The control of pigment migration in isolated erythrophores of Holocentrus ascensionis (Osbeck). II. The role of calcium. Cell. 1982 Jun;29(2):441–450. doi: 10.1016/0092-8674(82)90160-x. [DOI] [PubMed] [Google Scholar]
  18. Luby K. J., Porter K. R. The control of pigment migration in isolated erythrophores of Holocentrus ascensionis (Osbeck). I. Energy requirements. Cell. 1980 Aug;21(1):13–23. doi: 10.1016/0092-8674(80)90110-5. [DOI] [PubMed] [Google Scholar]
  19. Lynch T. J., Wu B. Y., Taylor J. D., Tchen T. T. Regulation of pigment organelle translocation. II. Participation of a cAMP-dependent protein kinase. J Biol Chem. 1986 Mar 25;261(9):4212–4216. [PubMed] [Google Scholar]
  20. Matthies H. J., Miller R. J., Palfrey H. C. Calmodulin binding to and cAMP-dependent phosphorylation of kinesin light chains modulate kinesin ATPase activity. J Biol Chem. 1993 May 25;268(15):11176–11187. [PubMed] [Google Scholar]
  21. McNiven M. A., Porter K. R. Chromatophores--models for studying cytomatrix translocations. J Cell Biol. 1984 Jul;99(1 Pt 2):152s–158s. doi: 10.1083/jcb.99.1.152s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McNiven M. A., Ward J. B. Calcium regulation of pigment transport in vitro. J Cell Biol. 1988 Jan;106(1):111–125. doi: 10.1083/jcb.106.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Miller P., Walter U., Theurkauf W. E., Vallee R. B., De Camilli P. Frozen tissue sections as an experimental system to reveal specific binding sites for the regulatory subunit of type II cAMP-dependent protein kinase in neurons. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5562–5566. doi: 10.1073/pnas.79.18.5562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mori M., Miki-Noumura T. Inhibition of gliding movement by calcium in doublet microtubules on Tetrahymena ciliary dyneins in vitro. Exp Cell Res. 1992 Dec;203(2):483–487. doi: 10.1016/0014-4827(92)90024-3. [DOI] [PubMed] [Google Scholar]
  25. Murphy D. B., Tilney L. G. The role of microtubules in the movement of pigment granules in teleost melanophores. J Cell Biol. 1974 Jun;61(3):757–779. doi: 10.1083/jcb.61.3.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Palazzo R. E., Lynch T. J., Taylor J. D., Tchen T. T. cAMP-independent and cAMP-dependent protein phosphorylations by isolated goldfish xanthophore cytoskeletons: evidence for the association of cytoskeleton with a carotenoid droplet protein. Cell Motil Cytoskeleton. 1989;13(1):21–29. doi: 10.1002/cm.970130104. [DOI] [PubMed] [Google Scholar]
  27. Rodionov V. I., Gyoeva F. K., Gelfand V. I. Kinesin is responsible for centrifugal movement of pigment granules in melanophores. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4956–4960. doi: 10.1073/pnas.88.11.4956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rozdzial M. M., Haimo L. T. Bidirectional pigment granule movements of melanophores are regulated by protein phosphorylation and dephosphorylation. Cell. 1986 Dec 26;47(6):1061–1070. doi: 10.1016/0092-8674(86)90821-4. [DOI] [PubMed] [Google Scholar]
  29. Rozdzial M. M., Haimo L. T. Reactivated melanophore motility: differential regulation and nucleotide requirements of bidirectional pigment granule transport. J Cell Biol. 1986 Dec;103(6 Pt 2):2755–2764. doi: 10.1083/jcb.103.6.2755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sammak P. J., Adams S. R., Harootunian A. T., Schliwa M., Tsien R. Y. Intracellular cyclic AMP not calcium, determines the direction of vesicle movement in melanophores: direct measurement by fluorescence ratio imaging. J Cell Biol. 1992 Apr;117(1):57–72. doi: 10.1083/jcb.117.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sanderson M. J., Charles A. C., Dirksen E. R. Mechanical stimulation and intercellular communication increases intracellular Ca2+ in epithelial cells. Cell Regul. 1990 Jul;1(8):585–596. doi: 10.1091/mbc.1.8.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sato-Yoshitake R., Yorifuji H., Inagaki M., Hirokawa N. The phosphorylation of kinesin regulates its binding to synaptic vesicles. J Biol Chem. 1992 Nov 25;267(33):23930–23936. [PubMed] [Google Scholar]
  33. Schliwa M., Bereiter-Hahn J. Pigment movements in fish melanophores: morphological and physiological studies. 3. The effects of colchicine and vinblastine. Z Zellforsch Mikrosk Anat. 1973 Dec 31;147(1):127–148. doi: 10.1007/BF00306604. [DOI] [PubMed] [Google Scholar]
  34. Schliwa M. Permeabilized cell models for the study of granule transport in pigment cells. Pigment Cell Res. 1987;1(2):65–68. doi: 10.1111/j.1600-0749.1987.tb00391.x. [DOI] [PubMed] [Google Scholar]
  35. Stearns M. E., Binder L. I. Evidence that MAP-2 may be involved in pigment granule transport in squirrel fish erythrophores. Cell Motil Cytoskeleton. 1987;7(3):221–234. doi: 10.1002/cm.970070305. [DOI] [PubMed] [Google Scholar]
  36. Swanson J. A. Pure thoughts with impure proteins: permeabilized cell models of organelle motility. Bioessays. 1993 Nov;15(11):715–722. doi: 10.1002/bies.950151104. [DOI] [PubMed] [Google Scholar]
  37. Tan J. L., Ravid S., Spudich J. A. Control of nonmuscle myosins by phosphorylation. Annu Rev Biochem. 1992;61:721–759. doi: 10.1146/annurev.bi.61.070192.003445. [DOI] [PubMed] [Google Scholar]
  38. Thaler C. D., Haimo L. T. Control of organelle transport in melanophores: regulation of Ca2+ and cAMP levels. Cell Motil Cytoskeleton. 1992;22(3):175–184. doi: 10.1002/cm.970220305. [DOI] [PubMed] [Google Scholar]
  39. Thaler C. D., Haimo L. T. Regulation of organelle transport in melanophores by calcineurin. J Cell Biol. 1990 Nov;111(5 Pt 1):1939–1948. doi: 10.1083/jcb.111.5.1939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Theurkauf W. E., Vallee R. B. Molecular characterization of the cAMP-dependent protein kinase bound to microtubule-associated protein 2. J Biol Chem. 1982 Mar 25;257(6):3284–3290. [PubMed] [Google Scholar]
  41. Trinczek B., Schwoch G. Immunoelectron microscopical localization of the catalytic subunit of cAMP-dependent protein kinases in brain microtubules and neurofilaments. FEBS Lett. 1990 Dec 17;277(1-2):167–170. doi: 10.1016/0014-5793(90)80835-7. [DOI] [PubMed] [Google Scholar]

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

RESOURCES