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  • Articles: DFG German National Licenses  (4)
  • Autoradiography  (2)
  • Opsin immunoreactivity  (2)
  • 1
    ISSN: 1432-0878
    Keywords: Pineal organ ; Retina ; Photoreceptors ; Photopigment ; Immunocytochemistry ; HPLC ; Autoradiography ; Mouse (C57BL)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: 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.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0878
    Keywords: Pineal organ (Meriones unguiculatus) ; Vasoactive intestinal polypeptide (VIP)-immunoreactive nervefibres ; VIP receptors ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of the indirect peroxidase-antiperoxidase immunohistochemical technique, nerve terminals exhibiting vasoactive intestinal polypeptide (VIP)-like-immunoreactivity were demonstrated in the pineal gland of the Mongolian gerbil (Meriones unguiculatus). Incubation of the superficial pineal gland of the gerbil with 60 pM of 125I-VIP showed that the gland exhibited saturation kinetics, and about 80% of the bound 125I-VIP could be displaced by adding a surplus of cold VIP. Incubation of unfixed, 0.1% and 4% paraformaldehyde-fixed cryostat sections of the gerbil forebrain with 125I-VIP also exhibited saturation kinetics, and displacement was possible by adding a surplus of the cold tracer. Receptor autoradiography on cryostat sections that had been incubated for 60 min with 125I-VIP showed a large number of grains over cortical areas, especially over the pyramidal layer of the hippocampus and the granular layer of the dentate gyrus. A prominent labelling of the pineal gland was also observed. The presence of VIP-like-immunoreactive nerve terminals and receptors for this molecule in the pineal gland of the Mongolian gerbil supports the biochemical studies demonstrating a stimulatory function of this molecule in the synthesis of melatonin.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0878
    Keywords: Pineal organ ; Parapineal organ ; Opsin immunoreactivity ; Cyclostome (Lampetra fluviatilis) ; Teleosts (Anguilla anguilla, Salmo gairdneri)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pineal complex of Lampetra fluviatilis, Anguilla anguilla and Salmo gairdneri was studied by means of the indirect immunohistochemical antiopsin reaction. Opsin-immunoreactive material was demonstrated in the outer segments of the photoreceptor cells in the pineal organ of all three species investigated. In the lamprey, the opsin-positive outer segments were located in the lumen of the pineal vesicle and atrium. In the two teleost species, the immunoreactive outer segments were observed in abundance in the pineal end-vesicle and stalk. These structures were found to accumulate in the prominent initial portion of the pineal stalk of the eel. In the rainbow trout, immunoreactive outer segments occurred in the wide orifice of the pineal recess at the roof of the third ventricle. In addition, outer segments of photoreceptor cells of the parapineal organ (“parapinealocytes”) displayed opsin immunoreactivity. In the lamprey, opsin immunoreactivity was restricted to the central portion of the ventral parapineal retina, while the parapinealocytes in the lateral portions did not bind the antibody. In the two teleosts, immunoreactive outer segments displayed a scattered pattern. These immunocytochemical results provide direct evidence that the photosensitivity of the pineal demonstrated electrophysiologically in lampreys and teleosts (cf. Dodt 1973) is based on an opsin-containing photopigment. The presence of opsin in cells of the parapineal organ strengthens the view that also this organ may be capable of direct light perception. In the lamprey, the exclusive opsin immunoreactivity of a circumscribed group of parapineal cells suggests the existence of two types of parapinealocytes. The significance of opsin-containing photoreceptor outer segments occurring in the most proximal portion of the teleost pineal stalk is discussed, especially with regard to the interpretation of results obtained from pinealectomy experiments.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0878
    Keywords: Pineal photoreceptors ; Opsin immunoreactivity ; Pineal neurons ; Acetylcholinesterase reaction ; Parapineal organ ; Teleosts (Phoxinus phoxinus L.)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pineal complex of the teleost, Phoxinus phoxinus L., was studied light-microscopically by the use of the indirect immunocytochemical antiopsin reaction and the histochemical acetylcholinersterase (AChE) method. Opsin-immunoreactive outer segments of photoreceptor cells were demonstrated in large numbers in all divisions of the pineal end-vesicle and in the pineal stalk. Moreover, they were found in the roof of the third ventricle, adjacent to the orifice of the pineal recess as well as scattered in the parapineal organ. These immunocytochemical observations provide direct evidence of the presence of an opsin associated with a photopigment in the photosensory cells of the pineal and parapineal organs of Phoxinus. By means of the AChE reaction (Karnovsky and Roots 1964) inner segments of pineal photoreceptors, intrinsic nerve cells, several intrapineal bundles of nerve fibers, and a prominent pineal tract were specifically marked. The pineal neurons can be divided into two types: one is located near the pineal lumen, the other near the basal lamina. The latter perikarya bear stained processes directed toward the photoreceptor layer. A rostral aggregation of two types of AChE-positive nerve cells occurs in the ventral wall of the pineal end-vesicle. The main portion of the AChE-positive pineal tract, which lies within the dorsal wall of the pineal stalk, can be followed to the posterior commissure where some of the nerve fibers course laterally. A few AChE-positive pineal nerve fibers run toward the lateral habenular nucleus via the habenular commissure. In the region of the subcommissural organ single AChE-positive neurons accompany the pineal tract. The nerve cells of the parapineal organ exhibit a moderate AChE activity. These findings extend the structural basis for the remarkable light-dependent activity of the pineal organ of Phoxinus phoxinus.
    Type of Medium: Electronic Resource
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