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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 29 (1979), S. 251-256 
    ISSN: 1432-0827
    Keywords: Cathepsin ; Calcification ; Dentinogenesis ; Proteoglycans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Cathepsin D was purified from rat liver using a new affinity chromatographic method, based on the coupling to the specific inhibitor pepstatin. This preparation was used for the production of specific antibodies from rabbit. The purified IgG fraction was conjugated to horseradish peroxidase in a two-step coupling procedure and used for electron microscopic immunohistochemistry of the odontoblast-predentine region of the rat incisor. Precipitates, indicating the presence of cathepsin D, were seen in the odontoblast, odontoblast process, and in the extracellular unmineralized matrix, the predentine. The observations are discussed in relation to proteoglycan degradation at the mineralization front simultaneous with crystal formation, and in relation to the function of lysosomal enzymes in the turnover of connective tissue.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 159 (1975), S. 233-243 
    ISSN: 1432-0878
    Keywords: Odontoblasts ; Predentine ; Dentine ; Calcification ; Scanning electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A scanning electron microscopic technique was used to investigate the surface structure of dentinogenically active odontoblasts. Thin pieces of rat incisors were fixed, rapidly frozen, freezedried at -70° C and fractured to expose new surfaces prior to examination in the SEM. Differences in the appearance of odontoblastic cell surfaces were seen, with the most extensive ridge formations at the distal part of the sides of the odontoblasts. The predentine area displayed a spongy structure which contrasted to the compact appearance of dentine. Results are discussed in relation to previous studies at the light microscopic and transmission electron microscopic levels.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 168 (1976), S. 277-287 
    ISSN: 1432-0878
    Keywords: Proteoglycans ; Odontoblasts, predentin, dentin ; Calcification ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of proteoglycans in the predentin of the rat incisor was investigated by ultrastructural histochemistry. Ruthenium red stained the cell coat of the odontoblasts as well as intracellular vesicles. There was also a staining of the extracellular matrix, but not of collagen fibers in the predentin. Treatment with the enzyme hyaluronidase prior to staining with ruthenium red abolished the staining of the vesicles and the extracellular matrix but not that of the cell coat. Bismuth nitrate and phosphotungstic acid gave similar staining of odontoblast vesicles and extracellular matrix. It is likely that the stained structures contain proteoglycans. The importance of these proteoglycans and their ultrastructural localization are discussed in relation to intracellular transport and the calcification process.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 129 (1972), S. 92-113 
    ISSN: 1432-0878
    Keywords: Neurosecretion ; Neurohypophysis ; Neurosecretory granule ; Rat ; Membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neurosecretory granules (NSG) of rat posterior pituitary glands were prepared by differential centrifugation techniques mainly according to the procedure as described by Barer, Heller and Lederis (1963). As revealed by electron microscopy, the recovery of neurophysin and the contents of enzymes, purified NSG were obtained in a pellet at 30 000 g/60 min (0.44 M sucrose). Eighteen h after injection of (35S) cysteine into the supraoptic nucleus 60% of the recovered radioactivity in the neural lobe was found in the NSG, whereas 20% was found in the final supernatant (100 000 g/120 min). Sixteen days after injection the NSG and the final supernatant fraction contained fairly equal amount of (35S) cysteine (approximately 40%). It is suggested that after a period of intragranular maturation neurophysin is extruded into an extragranular pool of neurosecretory material. With the use of conventional polyacrylamide-gel electrophoresis it was shown that the predominating proportion of radioactivity in the NSG after a hypothalamic injection of (35S) cysteine was located within the neurophysin fraction A and in fraction B. Fraction B is suggested to be partly bound to the NSG membranes. When the NSG soluble and NSG insoluble proteins, obtained after lysis of NSG, were separated on polyacrylamide gels in the presence of sodium dodecylsulphate, the highly radioactive soluble protein was shown to consist of two components with average molecular weights of 12 300 and 14 600. Most of the proteins in the lysate were found in the NSG membranes, though less radioactive. A component with a mol.wt. of 37 000 was enriched in the membrane fraction. At longer times after isotope injection the high mol.wt. proteins, particularly those of the NSG membranes, contained increased amounts of radioactivity.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0878
    Keywords: Supraoptic nucleus ; Neurohypophysis ; Neurosecretion ; Axonal transport ; Colchicine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of colchicine on the transport of proteins in the hypothalamo-neurohypophyseal tract of the rat was studied after injection of (35S) cysteine into the supraoptic nucleus (SON) region. Colchicine, dissolved in distilled water and administered subarachnoidally, inhibited the axonal transport of labelled proteins into the neurohypophysis: the radioactivity that was recovered in neurohypophyseal TCA precipitable material was markedly decreased and hardly any radioactivity was found in the neurohypophyseal proteins which were separated by polyacrylamide gel disc electrophoresis. As revealed by electron microscopy the SON cell bodies showed marked changes after treatment with colchicine: a deeply folded nucleolemma; a pronounced, granular nucleolus; a dispersed chromatin; a zonal distribution of cell organelles with mitochondria and lysosomes accumulated at the periphery, crowded ribosomes, often arranged as polyribosomes and richly branching short profiles of endoplasmic reticulum filled with filamentous material forming an inner perinuclear zone separated by enlarged Golgi complexes. The profiles of elongated Herring bodies in the infundibulum were increased. The axon terminals were filled with heavily osmiophilic neurosecretory granules. The neurofilaments were slightly or moderately increased in number. No apparent changes were observed with regard to the neurotubuli in the SON neurons. The glial cells of the supraopticoneurohypophyseal tract showed reactive changes with a proliferation of filamentous elements. The biochemical and ultrastructural findings are discussed especially with respect to the mechanisms of transport and release of neurosecretory granules.
    Type of Medium: Electronic Resource
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