Summary
We used microspectrophotometry, together with some associated electrophysiological measurements, to characterize the photopigments of R1–6 receptors in living, white-eyedDrosophila. Measurements were from the deep pseudopupil, an optical visualization of photopigment-containing rhabdomeres. Transmission changes associated with photointerconversion of rhodopsin (R480) and its stable metarhodopsin (M570) are easily seen (Fig. 1). Such transmission changes were measured to produce a difference specrum (Fig. 2); the isosbestic wavelength is about 502 nm. A photoequilibrium spectrum (Fig. 3), analyzed together with sensitivity data (Fig. 4a), was used to determine the fraction of M570 in photoequilibria established with various monochromatic wavelengths. From this, the quantum efficiency for M570 to R480 conversion relative to R480 to M570 efficiency was determined to be about 0.71. No dark regeneration of M570 to R480 occurred within a period of 60min (Fig. 4a, b).
The extent of conversion related to incident energy was estimated as a function of wavelength (Figs. 5 and 6). These experiments yielded the photosensitivity spectrum of the visual pigment (Fig. 6). Assuming that the absorption spectrum of rhodopsin (R480) is the same as the sensitivity spectrum as determined by electrophysiology, this photosensitivity spectrum was used to derive the spectrum of metarhodopsin (M570) (Fig. 7). The maximal extinction of M570 is about 1.43 times the maximal extinction of R480. The spectra of both states of the photopigment fit the Ebrey-Honig nomogram in the long wavelength band. Both of the photosensitivity spectra, measured in the living eye, associated with R480 and M570 respectively have a major UV maximum.
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Abbreviations
- ERG :
-
electroretinogram
- M orM570 :
-
metarhodopsin of R1–6
- NA :
-
numerical aperture
- PDA :
-
prolonged depolarizing afterpotential
- R orR480 :
-
rhodopsin of R1–6
- R1–6 :
-
retinula cells with peripheral rhabdomeres
- R7 andR8 :
-
retinula cells with central rhabdomeres
- SE :
-
standard error
- UV :
-
ultraviolet
- A, a, c, D, e, f, I, l. m, Q, S, t, α, γ, λ., Φ :
-
symbols used in quantitative treatment, see Methods
- bw, cn, norpA, sev, w :
-
Drosophila mutants, see Methods
References
Burkhardt, D.: Spectral sensitivity and other response characteristics of single cells in the arthropod eye. Symp. Soc. Exp. Biol.16, 86–109 (1962)
Burkhardt, D.: On the vision of insects. J. Comp. Physiol.120, 33–50 (1977)
Dartnall, H.J.A.: The interpretation of spectral sensitivity curves. Br. Med. Bull.9, 24–30 (1953)
Dartnall, H.J.A.: Photosensitivity. In: Handbook of sensory physiology, Vol. VII/I. Dartnall, H.J.A. (ed.), pp. 122–145. Berlin, Heidelberg, New York: Springer 1972
Dartnall, H.J.A., Goodeve, C.F., Lythgoe, R.J.: The quantitative analysis of the photochemical bleaching of visual purple solutions in monochromatic light. Proc. R. Soc. (London) Ser. A156, 158–170 (1936)
Doane, W.W.: Drosophila. In: Methods in developmental biology. Wilt, F.H., Wessells, N.K. (eds.). New York: Crowell 1967
Ebrey, T.G., Honig, B.: New wavelength dependent visual pigment nomograms. Vision Res.17, 147–151 (1977)
Eguchi, E., Waterman, T.H.: Freeze-etch and histochemical evidence for cycling in crayfish photoreceptor membranes. Cell Tissue Res.169, 419–434 (1976)
Fernandez, H.R., Nickel, E.E.: Ultrastructural and molecular characteristics of crayfish photoreceptor membranes. J. Cell Biol.69, 721–732 (1976)
Franceschini, N.: Sampling of the visual environment by the compound eye of the fly: Fundamentals and applications. In: Photoreceptor optics. Synder, A.W., Menzel, R. (eds.), pp. 98–125. Berlin, Heidelberg, New York: Springer 1975
Franceschini, N., Kirschfeld, K.: Les phénomènes de pseudopupille dans l'oeil composé deDrosophila. Kybernetik9, 159–182 (1971)
Franceschini, N., Kirschfeld, K.: Le contrôle automatique du flux lumineaux dans l'oeil composé des Diptères: Propriétés spectrales, statiques et dynamiques du mécanisme. Biol. Cybern.21, 181–203 (1976)
Hagins, W.A.: The visual process: Excitatory processes in primary receptor cells. Annu. Rev. Biophys. Bioeng.1, 131–158 (1972)
Hamdorf, K.: The physiology of invertebrate visual pigments. In: Handbook of sensory physiology, Vol. VII/6A. Autrum, H. (ed.), pp. 145–224. Berlin, Heidelberg, New York: Springer 1979
Hamdorf, K., Rosner, G.: Adaptation und Photoregeneration im Fliegenauge. J. Comp. Physiol.86, 281–292 (1973)
Hamdorf, K., Schwemer, J.: Photoregeneration and the adaptation process in insect photoreceptors. In: Photoreceptor optics. Snyder, A.W., Menzel, R. (eds.), pp. 263–289. Berlin, Heidelberg, New York: Springer 1975
Hamdorf, K., Paulsen, R., Schwemer, J.: Photoregeneration and sensitivity control of photoreceptors of invertebrates. In: Biochemistry and physiology of visual pigments. Langer, H. (ed.). Berlin, Heidelberg, New York: Springer 1973
Harris, W.A.: Color vision inDrosophila. Thesis for Ph.D. at California Institute of Technology (1976)
Harris, W.A., Ready, D.F., Lipson, E.D., Hudspeth, A.J., Stark, W.S.: Vitamin A deprivation andDrosophila photopigments. Nature (London)266, 648–650 (1977)
Harris, W.A., Stark, W.S.: Hereditary retinal degeneration inDrosophila melanogaster: a mutant defect associated with the phototransduction process. J. Gen. Physiol.69, 261–291 (1977)
Harris, W.A., Stark, W.S., Walker, J.A.: Genetic dissection of the photoreceptor system in the compound eye ofDrosophila melanogaster. J. Physiol. (London)256, 415–439 (1976)
Hecht, S., Shlaer, S., Pirenne, M.H.: Energy, quanta, and vision. J. Gen. Physiol.25, 819–840 (1942)
Hochstein, S., Minke, B., Hillman, P., Knight, B.W.: The kinetics of visual pigment systems I. Mathematical analysis. Biol. Cybern.30, 23–32 (1978)
Hoffmann, C., Langer, H.: Die spektrale Augenempfindlichkeit der Mutante “chalky” vonCalliphora erythrocephala. Naturwissenschaften48, 105 (1961)
Horridge, G.A., Mimura, K., Hardie, R.C.: Fly photoreceptors III. Angular sensitivity as a function of wavelength and the limits of resolution. Proc. R. Soc. (London) Ser. B194, 151–177 (1976)
Kirschfeld, K.: The function of photostable pigments in fly photoreceptors. Biophys. Struct. Mech.5, 177–128 (1979)
Kirschfeld, K., Franceschini, N.: Photostable pigments within the membrane of photoreceptors and their possible role. Biophys. Struct. Mech.3, 191–194 (1977)
Kirschfeld, K., Franceschini, N., Minke, B.: Evidence for a sensitizing pigment in fly photoreceptors. Nature (London)269, 386–390 (1977)
Kirschfeld, K., Feiler, R., Minke, B.: The kinetics of formation of metarhodopsin in intact photoreceptors of the fly. Z. Naturforsch., Teil C33, 1009–1010 (1978)
Lindsley, D.L., Grell, E.H.: Genetic variations ofDrosophila melanogaster. Oak Ridge, Tennessee: Oak Ridge National Laboratory 1968
Lo, M.-V.C.: Darkening of deep pseudopupil inDrosophila: An optical indication of inactivation of the peripheral photoreceptors. Ph.D. Thesis, Purdue University (1977)
Lo, M.-V.C., Pak, W.L.: Desensitization of peripheral photoreceptors shown by blue-induced decrease in transmittance ofDrosophila rhabdomeres. Nature (London)273, 772–774 (1978)
Minke, B., Kirschfeld, K.: The contribution of a sensitizing pigment to the photosensitivity spectra of fly rhodopsin and metarhodopsin. J. Gen. Physiol.73, 517–540 (1979)
Nickel, E., Menzel, R.: Insect UV- and green-photoreceptor membranes studied by the freeze-fracture technique. Cell Tissue Res.175, 357–368 (1976)
Ostroy, S.E.: Characteristics ofDrosophila rhodopsin in wild-type andnorpA vision transduction mutants. J. Gen. Physiol.72, 717–732 (1978)
Ostroy, S.E., Wilson, M., Pak, W.L.:Drosophila rhodopsin: photochemistry extraction and differences in thenorpA p12 phototransduction mutant. Biochem. Biophys. Res. Commun.59, 960–966 (1974)
Pak, W.L., Lidington, K.J.: Fast electrical potential from a long lived, long wavelength photoproduct of fly visual pigment. J. Gen. Physiol.63, 740–756 (1974)
Razmjoo, S., Hamdorf, K.: Visual sensitivity and the variation of total photopigment content in the blowfly photoreceptor membrane. J. Comp. Physiol.105, 279–286 (1976)
Stark, W.S.: Sensitivity and adaptation in R7, an ultraviolet photoreceptor, in theDrosophila retina. J. Comp. Physiol.115, 47–49 (1977)
Stark, W.S.: Spectral selectivity of visual response alterations mediated by interconversions of native and intermediate photopigments inDrosophila. J. Comp. Physiol.96, 343–356 (1975)
Stark, W.S., Zitzmann, W.G.: Isolation of adaptation mechanisms and photopigment spectra by vitamin A deprivation inDrosophila. J. Comp. Physiol.105, 15–27 (1976)
Stark, W.S., Ivanyshyn, A., Hu, K.G.: Spectral sensitivities and photopigments in adaptation of fly visual receptors. Naturwissenschaften63, 513–518 (1976)
Stark, W.S., Ivanyshyn, A.M., Greenberg, R.M.: Sensitivity and photopigments of R1–6, a two peaked photoreceptor, inDrosophila, Calliphora, andMusca. J. Comp. Physiol.121, 289–305 (1977)
Stark, W.S., Frayer, K.L., Johnson, M.A.: Photopigment and receptor properties inDrosophila compound eye and ocellar receptors. Biophys. Struct. Mech.5, 197–209 (1979a)
Stark, W.S., Kruizinga, B., Stavenga, D.G.: Rhabdomere fluorescence and UV vision in fly R1–6 receptors. Invest. Ophthalmol. Visual Sci. (Suppl.) p. 177 (Abstract) (1979b)
Stark, W.S., Stavenga, D.G., Kruizinga, B.: Fly photoreceptor fluorescence is related to UV sensitivity. Nature (London)270, 581–583 (1979c)
Stark, W.S., Sullivan, G., Hansen, K., Garfinkel, F.: Microspectrophotometry of dark revovery and bleaching ofDrosophila photopigments. In: AAAS 1980 Americal Association for the Advancement of Science (Abstract) (1980)
Stavenga, D.G.: Derivation of photochrome absorption spectra from absorbance difference measurements. Photochem. Photobiol.21, 105–110 (1975)
Stavenga, D.G.: Fly visual pigments difference in visual pigments of blowfly and dronefly peripheral retinula cells. J. Comp. Physiol.111, 137–152 (1976)
Stavenga, D.G.: Pseudopupils of compound eyes. In: Handbook of sensory physiology, Vol. VII/6A. Autrum, H. (ed.), pp. 358–439. Berlin, Heidelberg, New York: Springer 1979
Stavenga, D.G., Zantema, A., Kuiper, J.W.: Rhodopsin processes and the function of the pupil mechanism in flies. In: Biochemistry and physiology of visual pigments. Langer, H. (ed.). Berlin, Heidelberg, New York: Springer 1973
Stavenga, D.G., Flokstra, J.H., Kuiper, J.W.: Photopigment conversions expressed in the pupil mechanism of blowfly visual sense cells. Nature (London)253, 740–742 (1975)
Tan, K.E.W.P.: Vision in the ultraviolet. Ph. D. Thesis, Utrecht (1971)
Tsukahara, Y., Horridge, G.A.: Visual pigment spectra from sensitivity of single dronefly retinula cells. J. Comp. Physiol.114, 233–251 (1977)
White, R.H.: Insect visual pigments. Adv. Insect Physiol.13, 35–67 (1978)
Wijngaard, W., Stavenga, D.G.: On optical crosstalk between fly rhabdomeres. Biol. Cybern.18, 61–67 (1975)
Wright, R., Cosens, D.: Blue-adaptation and orange-adaptation in white-eyedDrosophila: Evidence that the prolonged after potential is correlated with the amount of M580 in R1–6. J. Comp. Physiol.113, 105–127 (1977)
Zuidervaart, H., Stavenga, D.G., Stark, W.S., Bernard, G.D.: Pupillary responses revealing receptor characteristics in wild-type and mutantDrosophila. In: Neuroscience abstracts (Society for Neuroscience Ninth Annual Meeting), 2755 (Abstract) (1979)
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Supported by grants BMS-74-12817 and BNS-76-11921 (NSF), 1-R01-EY02487-01A1 and 7-R01-EY-03408-01 (NIH), by locally administered funding at the University of Missouri and Johns Hopkins University and by a research fellowship from Rijksuniversiteit Groningen (the Netherlands) to William S. Stark. We extend special thanks to Drs. D. Stavenga and G. Bernard for comments on this manuscript and throughout this study. We thank Dr. D. Stavenga and Prof. J. Kuiper for their hospitality while working on part of this project in Groningen. We thank R. DeVoe, K. Frayer, F. Garfinkel, W. Green, R. Greenberg, K. Hansen, K. Hu, W. Koch, B. Kruizinga, J. Stackhouse, G. Sullivan, L. Warren, and H. Zuidervaart for technical assistance. We thank P. Ahl, N. Downer, R. Massof, B. Minke, and C.-L. Wey for substantive comments on earlier versions of this work.
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Stark, W.S., Johnson, M.A. Microspectrophotometry ofDrosophila visual pigments: Determinations of conversion efficiency in R1–6 receptors. J. Comp. Physiol. 140, 275–286 (1980). https://doi.org/10.1007/BF00606268
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DOI: https://doi.org/10.1007/BF00606268