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
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
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
Bridges CDB (1977) Rhodopsin regeneration in rod outer segments: utilization of 11-cis retinal and retinol. Exp Eye Res 24:571–580
Bridges CDB, Yoshikami S (1969) Uptake of tritiated retinaldehyde by the visual pigment of dark-adapted rats. Nature 221:275–276
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
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
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
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
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
Defoe DM, Bok D (1983) Rhodopsin chromophore exchanges among opsin molecules in the dark. Invest Ophthalmol Vis Sci 24:1211
Dohlman HG, Caron MG, Lefkowitz RJ (1987) Structure and function of the β2-adrenergic receptor-homology with rhodopsin. Kidney Int 23 [Suppl]:S2–13
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
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
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
Goodman de Witt S (1980) Plasma retinol-binding protein. Ann NY Acad Sci 348:378–388
Goodman de Witt S (1981) Retinoid-binding proteins in plasma and in cells. Ann NY Acad Sci 359:69–78
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
Grip WJ de, Daemen FJM, Bonting SL (1980) Isolation and purification of bovine rhodopsin. Methods Enzymol 67:301–320
Groenendijk GWI, Grip WJ de, Daemen FJM (1979) Identification and characterization of syn- and anti-isomers of retinaloximes. Anal Biochem 99:304–310
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
Hall MO, Bok D (1974) Incorporation of (3H) vitamin A into rhodopsin in light- and dark-adapted frogs. Exp Eye Res 18:105–117
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
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
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
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
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
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
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
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
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
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
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
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
Papermaster DS (1982) Preparation of antibodies to rhodopsin and the large protein of rod outer segments. Methods Enzymol 81:240–246
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
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
Pepperberg DR (1982) Generation of rhodopsin and “artificial” visual pigments in electrophysically active photoreceptors. Methods Enzymol 81:452–459
Pepperberg DR, Masland RH (1978) Retinal-induced sensitization of light-adapted rabbit photoreceptors. Brain Res 151:194–200
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
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
Phillips TS, Tsin ATC, Reiter RJ, Malsbury DW (1989) Retinoids in the bovine pineal gland. Brain Res Bull 22:259–261
Randerath K (1970) An evaluation of film detection methods for weak β-emitters, particularly tritium. Anal Biochem 34:188–205
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
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
Rogers AW (1979) Practical autoradiography. Review No. 20, Amersham, Arlington Heights, Ill
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
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
Sidman RL, Dowling JE (1963) Autoradiographic localization of 14C-vitamin A in dark-and light-adapted rat eyes. Anat Rec 145:286
Somers RL, Klein DC (1985) Rhodopsin kinase activity in the mammalian pineal gland and other tissues. Science 226:182–184
Sternberger LA (1979) Immunocytochemistry. Wiley, New York
Tabata M, Suzuki T, Niwa H (1985) Chromophores in the extraretinal photoreceptor (pineal organ) of teleosts. Brain Res 338:173–176
Ueck M (1968) Ultrastruktur des pinealen Sinnesapparates bei einigen Pipidae und Discoglossidae. Z Zellforsch 92:452–476
Wald G (1968) The molecular basis of visual excitation. Nature 219:800–807
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
Yau KW, Baylor DA (1989) Cyclic GMP-activated conductance of retinal photoreceptor cells. Annu Rev Neurosci 12:289–327
Yoshikami S, Nöll GN (1978) Isolated retinas synthesize visual pigments from retinol congeners delivered by liposomes. Science 200:1393–1395
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
Zimmerman WF, Daemen FJM, Bonting SL (1976) Distribution of enzyme activities in sub-cellular fractions of bovine retina. J Biol Chem 251:4700–4705
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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