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. 1982 Jun 1;93(3):849–859. doi: 10.1083/jcb.93.3.849

Ca2+-sequestering smooth endoplasmic reticulum in an invertebrate photoreceptor. II. Its properties as revealed by microphotometric measurements

PMCID: PMC2112163  PMID: 6288735

Abstract

Microphotometric measurements are used to investigate the functional properties of Ca2+-sequestering smooth endoplasmic reticulum (SER) in leech photoreceptors. 10-30 intact cells are mounted in a perfusion chamber, placed between crossed polarizers in a microphotometer, and permeabilized by saponin treatment. Subsequent perfusion with solutions containing Ca2+, MgATP, and oxalate leads to Ca uptake by SER. When the solubility product of Ca-oxalate is exceeded in the SER, birefringent Ca-oxalate precipitates form in the cisternae, leading to a large increase in the optical signal recorded from the preparation. The rate of increase in light intensity is used to measure the rate of Ca uptake. Ca uptake rate is linear with time over much of its course, can be switched on/off by the addition/withdrawal of Ca2+, ATP, or oxalate to/from the medium, and is inhibited by mersalyl and tetracaine. The Ca uptake mechanism has a high specificity for MgATP (KM,MgATP is approximately 0.8 mM). Uptake rates observed with dATP, GTP, UTP, ITP, and CTP are only 20-30% of the rate measured in ATP. The Ca pump has a high affinity for Ca2+ ions: the threshold for activation of the pump is approximately 5 x 10(-8) M, the apparent KM,Ca is approximately 4 x 10(-7) M. When Na+ or Li+ is substituted for K+, Ca uptake rate is decreased by 40-50%. The results show that the Ca2+-sequestering SER in leech photoreceptors shares some basic properties with skeletal muscle sarcoplasmic reticulum and supports the idea that certain subregions of the SER in invertebrate photoreceptors function as effective Ca2+ sinks/buffers close to the plasmalemma.

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Selected References

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  1. Bader C., Baumann F., Bertrand D. Role of intracellular calcium and sodium in light adaptation in the retina of the honey bee drone (Apis mellifera, L). J Gen Physiol. 1976 Apr;67(4):475–491. doi: 10.1085/jgp.67.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beil F. U., von Chak D., Hasselbach W., Weber H. H. Competition between oxalate and phosphate during active calcium accumulation by sarcoplasmic vesicles. Z Naturforsch C. 1977 Mar-Apr;32(3-4):281–287. doi: 10.1515/znc-1977-3-421. [DOI] [PubMed] [Google Scholar]
  3. Blaustein M. P., Ratzlaff R. W., Kendrick N. C., Schweitzer E. S. Calcium buffering in presynaptic nerve terminals. I. Evidence for involvement of a nonmitochondrial ATP-dependent sequestration mechanism. J Gen Physiol. 1978 Jul;72(1):15–41. doi: 10.1085/jgp.72.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Brown J. E., Blinks J. R. Changes in intracellular free calcium concentration during illumination of invertebrate photoreceptors. Detection with aequorin. J Gen Physiol. 1974 Dec;64(6):643–665. doi: 10.1085/jgp.64.6.643. [DOI] [PMC free article] [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. Calcium ion, a putative intracellular messenger for light-adaptation in Limulus ventral photoreceptors. Biophys Struct Mech. 1977 Jun 29;3(2):141–143. doi: 10.1007/BF00535809. [DOI] [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. Endo M., Iino M. Specific perforation of muscle cell membranes with preserved SR functions by saponin treatment. J Muscle Res Cell Motil. 1980 Mar;1(1):89–100. doi: 10.1007/BF00711927. [DOI] [PubMed] [Google Scholar]
  10. Fabiato A., Fabiato F. Calcium-induced release of calcium from the sarcoplasmic reticulum of skinned cells from adult human, dog, cat, rabbit, rat, and frog hearts and from fetal and new-born rat ventricles. Ann N Y Acad Sci. 1978 Apr 28;307:491–522. doi: 10.1111/j.1749-6632.1978.tb41979.x. [DOI] [PubMed] [Google Scholar]
  11. Fein A., Charlton J. S. A quantitative comparison of the effects of intracellular calcium injection and light adaptation on the photoresponse of Limulus ventral photoreceptors. J Gen Physiol. 1977 Nov;70(5):591–600. doi: 10.1085/jgp.70.5.591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ford G. D., Hess M. L. Calcium-accumulating properties of subcellular fractions of bovine vascular smooth muscle. Circ Res. 1975 Nov;37(5):580–587. doi: 10.1161/01.res.37.5.580. [DOI] [PubMed] [Google Scholar]
  13. HASSELBACH W., MAKINOSE M. [The calcium pump of the "relaxing granules" of muscle and its dependence on ATP-splitting]. Biochem Z. 1961;333:518–528. [PubMed] [Google Scholar]
  14. Hanani M., Shaw C. A potassium contribution to the response of the barnacle photoreceptor. J Physiol. 1977 Aug;270(1):151–163. doi: 10.1113/jphysiol.1977.sp011943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Johnson P. N., Inesi G. The effect of methylxanthines and local anesthetics on fragmented sarcoplasmic reticulum. J Pharmacol Exp Ther. 1969 Oct;169(2):308–314. [PubMed] [Google Scholar]
  16. Krebs W., Schaten B. The lateral photoreceptor of the barnacle, Balanus eburneus: quantitative morphology and fine structure. Cell Tissue Res. 1976 May 6;168(2):193–207. doi: 10.1007/BF00215877. [DOI] [PubMed] [Google Scholar]
  17. Lisman J. E., Brown J. E. Effects of intracellular injection of calcium buffers on light adaptation in Limulus ventral photoreceptors. J Gen Physiol. 1975 Oct;66(4):489–506. doi: 10.1085/jgp.66.4.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. MARTONOSI A., FERETOS R. SARCOPLASMIC RETICULUM. I. THE UPTAKE OF CA++ BY SARCOPLASMIC RETICULUM FRAGMENTS. J Biol Chem. 1964 Feb;239:648–658. [PubMed] [Google Scholar]
  20. Maaz G., Stieve H. The correlation of the receptor potential with the light induced transient increase in intracellular calcium-concentration measured by absorption change of arsenazo III injected into Limulus ventral nerve photoreceptor cell. Biophys Struct Mech. 1980;6(3):191–208. doi: 10.1007/BF00537293. [DOI] [PubMed] [Google Scholar]
  21. Mabuchi K., Sréter F. A. Use of cryostat sections for measurement of Ca2+ uptake by sarcoplasmic reticulum. Anal Biochem. 1978 Jun 1;86(2):733–742. doi: 10.1016/0003-2697(78)90801-1. [DOI] [PubMed] [Google Scholar]
  22. MacLennan D. H., Holland P. C. Calcium transport in sarcoplasmic reticulum. Annu Rev Biophys Bioeng. 1975;4(00):377–404. doi: 10.1146/annurev.bb.04.060175.002113. [DOI] [PubMed] [Google Scholar]
  23. Makinose M., Hasselbach W. Der Einfluss von Oxalat auf den Calcium-Transport isolierter Vesikel des sarkoplasmatischen Reticulum. Biochem Z. 1965 Dec 31;343(4):360–382. [PubMed] [Google Scholar]
  24. McGraw C. F., Somlyo A. V., Blaustein M. P. Localization of calcium in presynaptic nerve terminals. An ultrastructural and electron microprobe analysis. J Cell Biol. 1980 May;85(2):228–241. doi: 10.1083/jcb.85.2.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Moore L., Chen T., Knapp H. R., Jr, Landon E. J. Energy-dependent calcium sequestration activity in rat liver microsomes. J Biol Chem. 1975 Jun 25;250(12):4562–4568. [PubMed] [Google Scholar]
  26. NANNINGA L. B. The association constant of the complexes of adenosine triphosphate with magnesium, calcium, strontium, and barium ions. Biochim Biophys Acta. 1961 Dec 9;54:330–338. doi: 10.1016/0006-3002(61)90373-0. [DOI] [PubMed] [Google Scholar]
  27. PORTZEHL H., CALDWELL P. C., RUEEGG J. C. THE DEPENDENCE OF CONTRACTION AND RELAXATION OF MUSCLE FIBRES FROM THE CRAB MAIA SQUINADO ON THE INTERNAL CONCENTRATION OF FREE CALCIUM IONS. Biochim Biophys Acta. 1964 May 25;79:581–591. doi: 10.1016/0926-6577(64)90224-4. [DOI] [PubMed] [Google Scholar]
  28. Perrelet A., Bader C. R. Morphological evidence for calcium stores in photoreceptors of the honeybee drone retina. J Ultrastruct Res. 1978 Jun;63(3):237–243. doi: 10.1016/s0022-5320(78)80048-3. [DOI] [PubMed] [Google Scholar]
  29. Reuben J. P., Brandt P. W., Berman M., Grundfest H. Regulation of tension in the skinned crayfish muscle fiber. I. Contraction and relaxation in the absence of Ca (pCa is greater than 9). J Gen Physiol. 1971 Apr;57(4):385–407. doi: 10.1085/jgp.57.4.385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schmidt J. A., Fein A. Effects of calcium-blocking agents and phosphodiesterase inhibitors on voltage-dependent conductances in Limulus photoreceptors. Brain Res. 1979 Nov 2;176(2):369–374. doi: 10.1016/0006-8993(79)90991-0. [DOI] [PubMed] [Google Scholar]
  31. Sehilin J. Calcium uptake by subcellular fractions of pancreatic islets. Effects of nucleotides and theophylline. Biochem J. 1976 Apr 15;156(1):63–69. doi: 10.1042/bj1560063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sorenson M. M., Reuben J. P., Eastwood A. B., Orentlicher M., Katz G. M. Functional heterogeneity of the sarcoplasmic reticulum within sarcomeres of skinned muscle fibers. J Membr Biol. 1980 Mar 31;53(1):1–17. doi: 10.1007/BF01871168. [DOI] [PubMed] [Google Scholar]
  33. Tada M., Yamamoto T., Tonomura Y. Molecular mechanism of active calcium transport by sarcoplasmic reticulum. Physiol Rev. 1978 Jan;58(1):1–79. doi: 10.1152/physrev.1978.58.1.1. [DOI] [PubMed] [Google Scholar]
  34. Trotta E. E., de Meis L. ATP-dependent calcium accumulation in brain microsomes. Enhancement by phosphate and oxalate. Biochim Biophys Acta. 1975 Jun 25;394(2):239–247. doi: 10.1016/0005-2736(75)90262-x. [DOI] [PubMed] [Google Scholar]
  35. Walz B. Ca2+-sequestering smooth endoplasmic reticulum in an invertebrate photoreceptor. I. Intracellular topography as revealed by OsFeCN staining and in situ Ca accumulation. J Cell Biol. 1982 Jun;93(3):839–848. doi: 10.1083/jcb.93.3.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Walz B. Subcellular calcium localization and AT0-dependent Ca2+-uptake by smooth endoplasmic reticulum in an invertebrate photoreceptor cell. An ultrastrucutral, cytochemical and X-ray microanalytical study. Eur J Cell Biol. 1979 Oct;20(1):83–91. [PubMed] [Google Scholar]
  37. de Meis L., Rubin-Altschul M., Machado R. D. Comparative data of Ca2+ transport in brain and skeletal muscle microsomes. J Biol Chem. 1970 Apr 25;245(8):1883–1889. [PubMed] [Google Scholar]

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