Journal of Comparative Physiology A

, Volume 156, Issue 2, pp 213–222 | Cite as

An electron microprobe analysis of photoreceptors and outer pigment cells in the retina of the honeybee drone

  • J. A. Coles
  • R. Rick
Article

Summary

Intracellular concentrations of elements were measured in the retina of the honeybee drone,Apis mellifera ♂ by electron microprobe X-ray analysis of frozen dried sections (Table 2). Before shock-freezing, slices of retina were superfused with Ringer solution, as in other work in which intracellular activities of Na+, K+ and Cl were measured with ion-selective microelectrodes. The results give no evidence for any binding or sequestering of these elements in the cells, with the possible exception of K in photoreceptors (Table 3). In the special case of Na in outer pigment cells,aNa and [Na] were measured in the same piece of tissue: Na was present at a high concentration (55 mmol/l) but, again, we calculate that it was all freely dissolved in the cell water.

It was estimated that the subrhabdomeric cisternae of the photoreceptors contained 2–3 mmol/l Ca; otherwise, their electrolyte composition was similar to that of the cytoplasm. [Na], [K] and [Cl] in the rhabdom were what would be expected if the spaces between the microvilli were filled with Ringer solution

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References

  1. Bader CR, Baumann F, Bertrand D (1976) Role of intracellular calcium and sodium in light adaptation in the retina of the honeybee drone (Apis mellifera L). J Gen Physiol 67:475–491Google Scholar
  2. Bauer R, Rick R (1978) Computer analysis of X-ray spectra (EDS) from thin biological specimens. X-ray Spectrometry 7:63–69Google Scholar
  3. Brown JE, Blinks JR (1974) Changes in intracellular free calcium concentration during illumination of invertebrate photoreceptors. Detection with aequorin. J Gen Physiol 64:643–665Google Scholar
  4. Brown JE, Coles JA (1979) Saturation of the response to light inLimulus ventral photoreceptor. J Physiol (Lond) 296:373–392Google Scholar
  5. Brown JE, Lisman JE (1975) Intracellular Ca modulates sensitivity and time scale inLimulus ventral photoreceptors. Nature 258:252–254Google Scholar
  6. Brown JE, Brown PK, Pinto LH (1977) Detection of light-induced changes of intracellular ionized calcium concentration inLimulus ventral photoreceptors using Arsenazo III. J Physiol (Lond) 267:299–320Google Scholar
  7. Coles JA, Orkand RK (1982) Sodium activity in drone photoreceptors. J Physiol (Lond) 332:16P-17PGoogle Scholar
  8. Coles JA, Orkand RK (1983) Modification of potassium movement through the retina of the drone (Apis mellifera ♂) by glial uptake. J Physiol (Lond) 340:157–174Google Scholar
  9. Coles JA, Orkand RK (1985) Changes in sodium activity during light stimulation in photoreceptors, glia and extracellular space in drone retina. J Physiol (Lond) (in press)Google Scholar
  10. Coles JA, Tsacopoulos M (1979) K+ activity in photoreceptors, glial cells and extracellular space in the drone retina: changes during photostimulation. J Physiol (Lond) 290:525–549Google Scholar
  11. Coles JA, Tsacopoulos M (1981) Ionic and possible metabolic interactions between sensory neurones and glial cells in the retina of the honeybee drone. J Exp Biol 95:75–92Google Scholar
  12. Coles JA, Orkand RK, Munoz JL (1983) When the photoreceptors in the retina of the honeybee drone are stimulated, K+ activity in the glial cells rises more than Na+ activity falls. Experientia 39:630Google Scholar
  13. Dörge A, Rick R, Gehring K, Thurau K (1978) Preparation of freeze-dried cryosections for quantitative X-ray microanalysis of electrolytes in biological soft tissues. Pflügers Arch 373:85–97Google Scholar
  14. Elder HY, Gray CC, Jardine AG, Chapman JN, Biddlecombe WH (1982) Optimum conditions for cryoquenching of small tissue blocks in liquid coolants. J Microsc 126:45–61Google Scholar
  15. Galvan M, Dörge A, Beck F, Rick R (1984) Intracellular electrolyte concentrations in rat sympathetic neurones measured with an electron microprobe. Pflügers Arch 400:274–279Google Scholar
  16. Jehl B, Bauer R, Dörge A, Rick R (1981) The use of propane/isopentane mixtures for rapid freezing of biological specimens. J Microsc 123:307–309Google Scholar
  17. Levy S, Fein A (1983) Light-evoked Ca2+ increase measured near the threshold of adaptation inLimulus ventral photoreceptors. Invest Ophthalmol Visual Sci [Suppl], 24:177Google Scholar
  18. Meryman HT (1966) Review of biological freezing. In: Meryman HT (ed) Cryobiology. Academic Press, New York, pp 1–114Google Scholar
  19. Munoz JL, Deyhimi F, Coles JA (1983) Silanization of glass in the making of ion-sensitive microelectrodes. J Neurosci Methods 8:231–247Google Scholar
  20. Orkand RK, Coles JA, Tsacopoulos M (1985) The role of glial cells in ion homeostasis in the retina of the honeybee drone (Apis mellifera ♂). In: Roitbak A (ed) Functions of neuroglia. Proc Symposium Tiflis, 20–23 Nov. 1984 (in press)Google Scholar
  21. Perrelet A (1970) The fine structure of the retina of the honeybee drone. Z Zellforsch Mikrosk Anat 108:530–562Google Scholar
  22. Perrelet A, Bader CR (1978) Morphological evidence for calcium stores in photoreceptors of the honeybee drone retina. J Ultrastruct Res 63:237–243Google Scholar
  23. Perrelet A, Baumann F (1969) Evidence for extracellular space in the rhabdome of the honeybee drone eye. J Cell Biol 40:825–830Google Scholar
  24. Raggenbass M (1983) Effects of extracellular calcium and of light adaptation on the response to dim light in honeybee drone photoreceptors. J Physiol (Lond) 344:525–548Google Scholar
  25. Reynolds ES (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol 17:208–212Google Scholar
  26. Rick R, Dörge A, Thurau K (1982) Quantitative analysis of electrolytes in frozen dried sections. J Microsc 125:239–247Google Scholar
  27. Schlue WR, Wuttke W (1983) Potassium activity in leech neuropile glial cells changes with external potassium concentration. Brain Res 270:368–372Google Scholar
  28. Shaw SR (1977) Restricted diffusion and extracellular space in the insect retina. J Comp Physiol 113:257–282Google Scholar
  29. Skalska-Rakowska JM, Baumgartner B (1985) Longitudinal continuity of the subrhabdomeric cisternae in the photoreceptors of the compound eye of the drone,Apis mellifera. Experientia (in press)Google Scholar
  30. Somlyo AP, Walz B (1985) Elemental distribution inRana pipiens retinal rods: quantitative electron probe analysis. J Physiol (Lond) 258:188–195Google Scholar
  31. Somlyo AP, Somlyo AV, Shuman H (1979) Electron probe analysis of vascular smooth muscle. J Cell Biol 81:316–335Google Scholar
  32. Spurr AR (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–45Google Scholar
  33. Staples BR (1971) Certificate for standard reference material 2201, sodium chloride. Office of Standard Reference Materials, US National Bureau of StandardsGoogle Scholar
  34. Steiner RA, Oehme M, Ammann D, Simon W (1979) Neutral carrier sodium ion-selective microelectrode for intracellular studies. Analyt Chem 51:351–353Google Scholar
  35. Taylor PS, Thomas RC (1984) The effect of leakage on microelectrode measurements of intracellular sodium activity in crab muscle fibres. J Physiol (Lond) 352:539–550Google Scholar
  36. Walz B (1982a) Calcium-sequestering smooth endnoplasmic reticulum in retinula cells of the blowfly. J Ultrastruct Res 81:240–248Google Scholar
  37. Walz B (1982b) 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 93:839–848Google Scholar
  38. Walz B (1982c) Ca2+-sequestering smooth endoplasmic reticulum in an invertebrate photoreceptor. II. Its properties as revealed by microphotometric measurements. J Cell Biol 93:849–859Google Scholar
  39. Walz B, Somlyo AP (1984) Quantitative electron probe microanalysis of leech photoreceptors. J Comp Physiol A 154:81–87Google Scholar
  40. White RH, Michaud NA (1980) Calcium is a component of ommochrome pigment granules in insect eyes. Comp Biochem Physiol 65A:239–242Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • J. A. Coles
    • 1
  • R. Rick
    • 2
  1. 1.Laboratoire d'ophtalmologie expérimentaleUniversité de GenèveGenève 4Switzerland
  2. 2.Physiologisches InstitutUniversität MünchenMünchen 2Federal Republic of Germany

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