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. 1994 Jun 1;125(5):1119–1125. doi: 10.1083/jcb.125.5.1119

On the localization of voltage-sensitive calcium channels in the flagella of Chlamydomonas reinhardtii

PMCID: PMC2120057  PMID: 8195293

Abstract

This study was undertaken to prove that voltage-sensitive calcium channels controlling the photophobic stop response of the unicellular green alga Chlamydomonas reinhardtii are exclusively found in the flagellar region of the cell and to answer the question as to their exact localization within the flagellar membrane. The strategy used was to amputate flagella to a variable degree without perturbing the electrical properties of the cell and measure flagellar currents shortly after amputation and during the subsequent regeneration process. Under all conditions, a close correlation was found between current size and flagellar length, strongly suggesting that the channels that mediate increases in intraflagellar calcium concentration are confined to and distributed over the total flagellar length. Bald mutants yielded tiny flagellar currents, in agreement with the existence of residual flagellar stubs. In the presence of the protein synthesis inhibitor cycloheximide, flagellar length and flagellar currents also recovered in parallel. Recovery came to an earlier end, however, leveling off at a time when in the absence of cycloheximide only half maximal values were achieved. This suggests the existence of a pool of precursors, which permits the maintenance of a constant ratio between voltage-sensitive calcium channels and other intraflagellar proteins.

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Selected References

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

  1. Baylor D. A., Lamb T. D., Yau K. W. The membrane current of single rod outer segments. J Physiol. 1979 Mar;288:589–611. [PMC free article] [PubMed] [Google Scholar]
  2. Beckmann M., Hegemann P. In vitro identification of rhodopsin in the green alga Chlamydomonas. Biochemistry. 1991 Apr 16;30(15):3692–3697. doi: 10.1021/bi00229a014. [DOI] [PubMed] [Google Scholar]
  3. Bessen M., Fay R. B., Witman G. B. Calcium control of waveform in isolated flagellar axonemes of Chlamydomonas. J Cell Biol. 1980 Aug;86(2):446–455. doi: 10.1083/jcb.86.2.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cheshire J. L., Keller L. R. Uncoupling of Chlamydomonas flagellar gene expression and outgrowth from flagellar excision by manipulation of Ca2+. J Cell Biol. 1991 Dec;115(6):1651–1659. doi: 10.1083/jcb.115.6.1651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dunlap K. Localization of calcium channels in Paramecium caudatum. J Physiol. 1977 Sep;271(1):119–133. doi: 10.1113/jphysiol.1977.sp011993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fisher G., Kaneshiro E. S., Peters P. D. Divalent cation affinity sites in Paramecium aurelia. J Cell Biol. 1976 May;69(2):429–442. doi: 10.1083/jcb.69.2.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Foster K. W., Saranak J., Patel N., Zarilli G., Okabe M., Kline T., Nakanishi K. A rhodopsin is the functional photoreceptor for phototaxis in the unicellular eukaryote Chlamydomonas. Nature. 1984 Oct 25;311(5988):756–759. doi: 10.1038/311756a0. [DOI] [PubMed] [Google Scholar]
  8. Foster K. W., Smyth R. D. Light Antennas in phototactic algae. Microbiol Rev. 1980 Dec;44(4):572–630. doi: 10.1128/mr.44.4.572-630.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gilula N. B., Satir P. The ciliary necklace. A ciliary membrane specialization. J Cell Biol. 1972 May;53(2):494–509. doi: 10.1083/jcb.53.2.494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hegemann P., Hegemann U., Foster K. W. Reversible bleaching of Chlamydomonas reinhardtii rhodopsin in vivo. Photochem Photobiol. 1988 Jul;48(1):123–128. doi: 10.1111/j.1751-1097.1988.tb02796.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Imam S. H., Snell W. J. The Chlamydomonas cell wall degrading enzyme, lysin, acts on two substrates within the framework of the wall. J Cell Biol. 1988 Jun;106(6):2211–2221. doi: 10.1083/jcb.106.6.2211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jaenicke L., Kuhne W., Spessert R., Wahle U., Waffenschmidt S. Cell-wall lytic enzymes (autolysins) of Chlamydomonas reinhardtii are (hydroxy)proline-specific proteases. Eur J Biochem. 1987 Dec 30;170(1-2):485–491. doi: 10.1111/j.1432-1033.1987.tb13725.x. [DOI] [PubMed] [Google Scholar]
  14. Johnson K. A., Rosenbaum J. L. Flagellar regeneration in Chlamydomonas: a model system for studying organelle assembly. Trends Cell Biol. 1993 May;3(5):156–161. doi: 10.1016/0962-8924(93)90136-o. [DOI] [PubMed] [Google Scholar]
  15. Kamiya R., Hasegawa E. Intrinsic difference in beat frequency between the two flagella of Chlamydomonas reinhardtii. Exp Cell Res. 1987 Nov;173(1):299–304. doi: 10.1016/0014-4827(87)90357-0. [DOI] [PubMed] [Google Scholar]
  16. Kamiya R., Witman G. B. Submicromolar levels of calcium control the balance of beating between the two flagella in demembranated models of Chlamydomonas. J Cell Biol. 1984 Jan;98(1):97–107. doi: 10.1083/jcb.98.1.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lefebvre P. A., Nordstrom S. A., Moulder J. E., Rosenbaum J. L. Flagellar elongation and shortening in Chlamydomonas. IV. Effects of flagellar detachment, regeneration, and resorption on the induction of flagellar protein synthesis. J Cell Biol. 1978 Jul;78(1):8–27. doi: 10.1083/jcb.78.1.8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Litvin F. F., Sineshchekov O. A., Sineshchekov V. A. Photoreceptor electric potential in the phototaxis of the alga Haematococcus pluvialis. Nature. 1978 Feb 2;271(5644):476–478. doi: 10.1038/271476a0. [DOI] [PubMed] [Google Scholar]
  19. Machemer H., Ogura A. Ionic conductances of membranes in ciliated and deciliated Paramecium. J Physiol. 1979 Nov;296:49–60. doi: 10.1113/jphysiol.1979.sp012990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Matsuda Y., Saito T., Yamaguchi T., Kawase H. Cell wall lytic enzyme released by mating gametes of Chlamydomonas reinhardtii is a metalloprotease and digests the sodium perchlorate-insoluble component of cell wall. J Biol Chem. 1985 May 25;260(10):6373–6377. [PubMed] [Google Scholar]
  21. Morris C. E., Sigurdson W. J. Stretch-inactivated ion channels coexist with stretch-activated ion channels. Science. 1989 Feb 10;243(4892):807–809. doi: 10.1126/science.2536958. [DOI] [PubMed] [Google Scholar]
  22. Moss A. G., Tamm S. L. A calcium regenerative potential controlling ciliary reversal is propagated along the length of ctenophore comb plates. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6476–6480. doi: 10.1073/pnas.84.18.6476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nichols K. M., Rikmenspoel R. Control of flagellar motion in Chlamydomonas and Euglena by mechanical microinjection of Mg2+ and Ca2+ and by electric current injection. J Cell Sci. 1978 Feb;29:233–247. doi: 10.1242/jcs.29.1.233. [DOI] [PubMed] [Google Scholar]
  24. Olesen S. P., Clapham D. E., Davies P. F. Haemodynamic shear stress activates a K+ current in vascular endothelial cells. Nature. 1988 Jan 14;331(6152):168–170. doi: 10.1038/331168a0. [DOI] [PubMed] [Google Scholar]
  25. Roberts W. M. Spatial calcium buffering in saccular hair cells. Nature. 1993 May 6;363(6424):74–76. doi: 10.1038/363074a0. [DOI] [PubMed] [Google Scholar]
  26. Rosenbaum J. L., Moulder J. E., Ringo D. L. Flagellar elongation and shortening in Chlamydomonas. The use of cycloheximide and colchicine to study the synthesis and assembly of flagellar proteins. J Cell Biol. 1969 May;41(2):600–619. doi: 10.1083/jcb.41.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanders M. A., Salisbury J. L. Centrin-mediated microtubule severing during flagellar excision in Chlamydomonas reinhardtii. J Cell Biol. 1989 May;108(5):1751–1760. doi: 10.1083/jcb.108.5.1751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schmidt J. A., Eckert R. Calcium couples flagellar reversal to photostimulation in Chlamydomonas reinhardtii. Nature. 1976 Aug 19;262(5570):713–715. doi: 10.1038/262713a0. [DOI] [PubMed] [Google Scholar]
  29. Sueoka N., Chiang K. S., Kates J. R. Deoxyribonucleic acid replication in meiosis of Chlamydomonas reinhardi. I. Isotopic transfer experiments with a strain producing eight zoospores. J Mol Biol. 1967 Apr 14;25(1):47–66. doi: 10.1016/0022-2836(67)90278-1. [DOI] [PubMed] [Google Scholar]
  30. Tamm S. L. Iontophoretic localization of Ca-sensitive sites controlling activation of ciliary beating in macrocilia of Beroë: the ciliary rete. Cell Motil Cytoskeleton. 1988;11(2):126–138. doi: 10.1002/cm.970110206. [DOI] [PubMed] [Google Scholar]
  31. Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]
  32. Uhl R., Hegemann P. Probing visual transduction in a plant cell: Optical recording of rhodopsin-induced structural changes from Chlamydomonas reinhardtii. Biophys J. 1990 Nov;58(5):1295–1302. doi: 10.1016/S0006-3495(90)82469-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Watson P. A. Direct stimulation of adenylate cyclase by mechanical forces in S49 mouse lymphoma cells during hyposmotic swelling. J Biol Chem. 1990 Apr 25;265(12):6569–6575. [PubMed] [Google Scholar]

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