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Rod-opsin immunoreaction in the pineal organ of the pigmented mouse does not indicate the presence of a functional photopigment

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Abstract

The aim of the present study was to characterize the rod-opsin immunoreaction in the mammalian pineal organ. Pigmented mice (strain C57BL) were selected as the animal model. Immunocytochemical investigations involving the use of highly specific polyclonal and monoclonal antibodies against bovine rod-opsin (the apoprotein of the photopigment rhodopsin) showed that approximately 25% of all pinealocytes were rod-opsin immunoreactive. Immunoblotting techniques revealed three protein bands of approximately 40, 75, and 110 kDa; these were detected by the monoclonal antibody and the polyclonal antiserum in retinal and pineal extracts. These protein bands presumably represented the monomeric, dimeric and trimeric forms of rod-opsin. The amount of rod-opsin in retina and pineal organ was quantified by means of an enzyme-linked immunosorbent assay. This yielded 570±30 pmoles rod-opsin per eye and 0.3±0.05 pmoles rod-opsin per pineal organ. High pressure liquid chromatography analysis of whole eye extracts demonstrated the chromophoric group of the photopigment rhodopsin, 11-cis retinal, and its isomer, all-trans-retinal. A shift from 11-cis retinal to all-trans-retinal was found upon light adaptation. No retinals were detected in the pineal organ. Autoradiographic investigations showed that 3H-retinol, intraperitoneally injected into the animals, was incorporated into the outer and inner segments of retinal photoreceptors, but not into the pineal organ. It is concluded that the mouse pineal organ contains the authentic apoprotein of rhodopsin but that it lacks retinal derivatives as essential components of all known vertebrate photopigments. Consequently, the “photoreceptor-specific” proteins of the mammalian pineal organ are not involved in photoreception and phototransduction, but may serve other functions to be explored in future studies.

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References

  • Abe T, Shinohara T (1990) S-antigen from the rat retina and pineal gland have identical sequences. Exp Eye Res 51:111

    Google Scholar 

  • Adamus G, Zam ZS, Arendt A, Palczewski K, McDowell JH, Hargrave PA (1991) Anti-rhodopsin monoclonal antibodies of defined specificity; characterization and application. Vision Res 31:17–31

    Google Scholar 

  • Bridges CDB (1976) Vitamin A and the role of the pigment epithelium during bleaching and regeneration of rhodopsin in the frog eye. Exp Eye Res 22:435–455

    Google Scholar 

  • Bridges CDB (1977) Rhodopsin regeneration in rod outer segments: utilization of 11-cis retinal and retinol. Exp Eye Res 24:571–580

    Google Scholar 

  • Bridges CDB, Yoshikami S (1969) Uptake of tritiated retinaldehyde by the visual pigment of dark-adapted rats. Nature 221:275–276

    Google Scholar 

  • Bridges CDB, Landers RA, Fong SL, Liou GI (1986) Interstitial retinol-binding protein (IRBP) in rat and bovine pineal organs: evolutionary vestige or functional molecule? In: O'Brien P, Klein DC (eds) Pineal and retinal relationships. Academic Press, New York, pp 383–400

    Google Scholar 

  • Bridges CDB, Foster RG, Landers RA, Liou GI, Fong SL (1987) Interstitial retinol-binding protein and cellular retinal-binding protein in the mammalian pineal. Vision Res 27:2049–2060

    Google Scholar 

  • Burnette WN (1981) Electrophoresis transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203

    Google Scholar 

  • Chabre M, Applebury ML (1986) Interaction of photoactivated rhodopsin with photoreceptor proteins: the cGMP cascade. In: Stieve H (ed) The molecular mechanism of photoreception. Dahlem Konferenzen 1986. Springer, Berlin Heidelberg New York, pp 51–66

    Google Scholar 

  • Chader GJ, Wiggert B, Lai YL, Lee L, Fletcher RT (1983) Interphotoreceptor retinol-binding protein: a possible role in retinoid transport to the retina. In: Osborne NN, Chader GJ (eds) Progress in retinal research, vol 2. Pergamon Press, Oxford New York, pp 163–189

    Google Scholar 

  • Defoe DM, Bok D (1983) Rhodopsin chromophore exchanges among opsin molecules in the dark. Invest Ophthalmol Vis Sci 24:1211

    Google Scholar 

  • Dohlman HG, Caron MG, Lefkowitz RJ (1987) Structure and function of the β2-adrenergic receptor-homology with rhodopsin. Kidney Int 23 [Suppl]:S2–13

    Google Scholar 

  • Flight WFG, Donselaar E van (1975) Ultrastructural aspects of the incorporation of 3H-vitamin A in the pineal organ of the urodele, Diemictylus viridescens viridescens. Proc Kon Ned Akad Wet Ser C 78:130–142

    Google Scholar 

  • Foster RG, Schalken JJ, Timmers AM, Grip WJ de (1989a) A comparison of some photoreceptor characteristics in the pineal and retina. I. The Japanese quail (Coturnix coturnix). J Comp Physiol [A] 165:553–563

    Google Scholar 

  • Foster RG, Timmers AM, Schalken JJ, Grip WJ de (1989b) A comparison of some photoreceptor characteristics in the pineal and retinae. II. The Djungarian hamster (Phodopus sungorus). J Comp Physiol [A] 65:565–572

    Google Scholar 

  • Goodman de Witt S (1980) Plasma retinol-binding protein. Ann NY Acad Sci 348:378–388

    Google Scholar 

  • Goodman de Witt S (1981) Retinoid-binding proteins in plasma and in cells. Ann NY Acad Sci 359:69–78

    Google Scholar 

  • Grip WJ de (1985) Immunochemistry of rhodopsin. In: Osborne NN, Chader GJ (eds) Progress in retinal research, vol 4. Pergamon Press, Oxford New York, pp 137–180

    Google Scholar 

  • Grip WJ de, Daemen FJM, Bonting SL (1980) Isolation and purification of bovine rhodopsin. Methods Enzymol 67:301–320

    Google Scholar 

  • Groenendijk GWI, Grip WJ de, Daemen FJM (1979) Identification and characterization of syn- and anti-isomers of retinaloximes. Anal Biochem 99:304–310

    Google Scholar 

  • Groenendijk GWI, Grip WJ de, Daemen FJM (1980) Quantitative determination of retinals with complete retention of their geometric configuration. Biochim Biophys Acta 617:430–438

    Google Scholar 

  • Hall MO, Bok D (1974) Incorporation of (3H) vitamin A into rhodopsin in light- and dark-adapted frogs. Exp Eye Res 18:105–117

    Google Scholar 

  • Hirosawa K, Yamada E (1973) The localization of the vitamin A in the mouse liver as revealed by electron microscope radiography. J Electron Microsc 22:337–346

    Google Scholar 

  • Ito T, Nemoto M (1952) Über die Kupfferschen Sternzellen und die “Fettspeicherungszellen” (“fat-storing cells”) in der Blutkapillarenwand der menschlichen Leber. Okajima Folia Anat Jpn 24:243–258

    Google Scholar 

  • Korf HW, Ekström P (1987) Photoreceptor differentiation and neuronal organization of the pineal organ. In: Trentini GP, Gaetani C de, Pévet P (eds) Fundamentals and clinics in pineal research. Raven Press, New York, pp 35–49

    Google Scholar 

  • Korf HW, Møller M, Gery I, Zigler JS, Klein DC (1985a) Immunocytochemical demonstration of retinal S-antigen in the pineal organ of four mammalian species. Cell Tissue Res 239:81–85

    Google Scholar 

  • Korf HW, Foster RG, Ekström P, Schalken JJ (1985b) Opsin-like immunoreaction in the retinae and pineal organs of four mammalian species. Cell Tissue Res 242:645–648

    Google Scholar 

  • Korf HW, Oksche A, Ekström P, Veen T van, Zigler JS, Gery I, Stein P, Klein DC (1986a) S-antigen immunocytochemistry. In: O'Brien P, Klein DC (eds) Pineal and retinal relationships. Academic Press, New York, pp 343–355

    Google Scholar 

  • Korf HW, Oksche A, Ekström P, Gery I, Zigler JS, Klein DC (1986b) Pinealocyte projections into the mammalian brain revealed with S-antigen antiserum. Science 231:735–737

    Google Scholar 

  • Korf HW, Sato T, Oksche A (1990) Complex relationships between the pineal organ and the medial habenular nucleus-pretectal region of the mouse as revealed by S-antigen immunocytochemistry. Cell Tissue Res 261:493–500

    Google Scholar 

  • Korf HW, Korf B, Schachenmayr W, Chader GJ, Wiggert B (1992) Immunocytochemical demonstration of interphotoreceptor retinoid-binding protein in cerebellar medulloblastoma. Acta Neuropathol (Berl) 83:482–487

    Google Scholar 

  • Kramm CM, Korf H-W, Czerwionka M, Schachenmayr W, Grip WJ de (1991) Photoreceptor differentiation in cerebellar medulloblastoma: evidence for a functional photopigment and authentic S-antigen (arrestin). Acta Neuropathol (Berl) 81:296–302

    Google Scholar 

  • Müller B, Peichl L, Grip WJ de, Gery I, Korf HW (1989) Opsin-and S-antigen-like immunoreactions in photoreceptors of the tree shrew retina. Invest Ophthalmol Vis Sci 30:530–535

    Google Scholar 

  • Palczewski K, Carruth ME, Adamus G, McDowell JH, Hargrave PA (1990) Molecular, enzymatic and functional properties of rhodopsin kinase from rat pineal gland. Vision Res 30:1129–1137

    Google Scholar 

  • Papermaster DS (1982) Preparation of antibodies to rhodopsin and the large protein of rod outer segments. Methods Enzymol 81:240–246

    Google Scholar 

  • Papermaster DS, Converse CA, Zorn M (1976) Biosynthesis and immunocytochemical characterization of a large protein in frog and cattle rod outer segment membranes. Exp Eye Res 23:105–116

    Google Scholar 

  • Papermaster DS, Schneider BG, Zorn MA, Kraehenbuhl JP (1978) Immunocytochemical localization of opsin in outer segments and Golgi zones of frog photoreceptor cells. An electron microscope analysis of cross-linked albumin-embedded retinas. J Cell Biol 77:196–210

    Google Scholar 

  • Pepperberg DR (1982) Generation of rhodopsin and “artificial” visual pigments in electrophysically active photoreceptors. Methods Enzymol 81:452–459

    Google Scholar 

  • Pepperberg DR, Masland RH (1978) Retinal-induced sensitization of light-adapted rabbit photoreceptors. Brain Res 151:194–200

    Google Scholar 

  • Pepperberg DR, Lurie M, Brown PK, Dowling JE (1976) Visual adaptation: effects of externally applied retinal on the lightadapted, isolated skate retina. Science 191:394–396

    Google Scholar 

  • Pepperberg DR, Brown PK, Lurie M, Dowling JE (1978) Visual pigment and photoreceptor sensitivity in the isolated skate retina. J Gen Physiol 71:369–396

    Google Scholar 

  • Phillips TS, Tsin ATC, Reiter RJ, Malsbury DW (1989) Retinoids in the bovine pineal gland. Brain Res Bull 22:259–261

    Google Scholar 

  • Randerath K (1970) An evaluation of film detection methods for weak β-emitters, particularly tritium. Anal Biochem 34:188–205

    Google Scholar 

  • Redmond TM, Wiggert B, Robey FA, Nguyen NY, Lewis MS, Lee L, Chader GJ (1986) Isolation and characterization of monkey interphotoreceptor retinoid-binding protein, a unique extracellular matrix component of the retina. Biochemistry 24:787–793

    Google Scholar 

  • Rodrigues MM, Hackett J, Gaskins R, Wiggert B, Lee L, Redmond M, Chader GJ (1986) Interphotoreceptor retinoid-binding protein in retinal rod cells and pineal gland. Invest Ophthalmol Vis Sci 27:844–850

    Google Scholar 

  • Rogers AW (1979) Practical autoradiography. Review No. 20, Amersham, Arlington Heights, Ill

    Google Scholar 

  • Schalken JJ (1987) The visual pigment rhodopsin: immunochemical aspects and induction of experimental autoimmune uveoretinitis. Thesis, University of Nijmegen

  • Sherman BS (1970) Autoradiographic localization of (3H)-retinol and derivatives in rat retina. Exp Eye Res 10:53–57

    Google Scholar 

  • Shinohara T, Craft CM, Stein P, Zigler JS, Wistow G, Katial A, Gery I, Klein DC (1986) Isolation of cDNAs for bovine S-antigen. In: O'Brien P, Klein DC (eds) Pineal and retinal relationships. Academic Press, Orlando, pp 331–342

    Google Scholar 

  • Sidman RL, Dowling JE (1963) Autoradiographic localization of 14C-vitamin A in dark-and light-adapted rat eyes. Anat Rec 145:286

    Google Scholar 

  • Somers RL, Klein DC (1985) Rhodopsin kinase activity in the mammalian pineal gland and other tissues. Science 226:182–184

    Google Scholar 

  • Sternberger LA (1979) Immunocytochemistry. Wiley, New York

    Google Scholar 

  • Tabata M, Suzuki T, Niwa H (1985) Chromophores in the extraretinal photoreceptor (pineal organ) of teleosts. Brain Res 338:173–176

    Google Scholar 

  • Ueck M (1968) Ultrastruktur des pinealen Sinnesapparates bei einigen Pipidae und Discoglossidae. Z Zellforsch 92:452–476

    Google Scholar 

  • Wald G (1968) The molecular basis of visual excitation. Nature 219:800–807

    Google Scholar 

  • Yamaki K, Tsuda M, Kikuchi T, Chen K-H, Huang K-P, Shinohara T (1990) Structural organization of the human S-antigen. J Biol Chem 265:20757–20762

    Google Scholar 

  • Yau KW, Baylor DA (1989) Cyclic GMP-activated conductance of retinal photoreceptor cells. Annu Rev Neurosci 12:289–327

    Google Scholar 

  • Yoshikami S, Nöll GN (1978) Isolated retinas synthesize visual pigments from retinol congeners delivered by liposomes. Science 200:1393–1395

    Google Scholar 

  • Zimmerman WF, Lion R, Daemen FJM, Bonting SL (1975) Distribution of specific retinol dehydrogenase activities in sub-cellular fractions of bovine retina and pigment epithelium. Exp Eye Res 21:325–332

    Google Scholar 

  • Zimmerman WF, Daemen FJM, Bonting SL (1976) Distribution of enzyme activities in sub-cellular fractions of bovine retina. J Biol Chem 251:4700–4705

    Google Scholar 

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Kramm, C.M., de Grip, W.J. & Korf, H.W. Rod-opsin immunoreaction in the pineal organ of the pigmented mouse does not indicate the presence of a functional photopigment. Cell Tissue Res 274, 71–78 (1993). https://doi.org/10.1007/BF00327987

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  • DOI: https://doi.org/10.1007/BF00327987

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