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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 146 (1975), S. 225-243 
    ISSN: 1432-0568
    Keywords: Differentiation ; Sympathetic ganglion ; Culture ; Electron microscopy ; Histochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Immature sympathetic ganglia prepared from 51/2- or 6-day-old chick embryos were cultured up to one month. The in vitro development was followed by phase microscopy, electron microscopy and using histochemistry for catecholamines, monoamine oxidase and cholinesterases. During the first week of culture extensive plexuses of nerve fibres were formed between and around the clusters of nerve cells. Mature-looking neurons were observed in the cultures by phase microscopy after three weeks, at which age the mean diameter of the perikarya was more than doubled. Varying catecholamine fluorescence was observed in the perikarya during the entire culture period. The nerve fibres showed usually only weak fluorescence, but, in the older cultures, bright varicosities were regularly found in the fibres. Monoamine oxidase activity was demonstrated already at three days of culture and the reaction was maintained positive. Weak or moderate acetyl-cholinesterase activity was demonstrated in the sympathicoblasts and young sympathetic neurons and their processes. The axolemma showed acetylcholinesterase activity also around the nerve terminals containing small dense cored vesicles. Reactions for the non-specific cholinesterases were negative. Electron microscopy of the 30-day-old cultures revealed that the clusters of nerve cells consisted of mature sympathetic neurons, which contained large (60–200 nm) and small (35–60 nm) granular catecholamine-storing vesicles. Glial cells were almost totally lacking. Large numbers of nerve terminals containing both large and small granular vesicles were observed in the clusters, often in synaptic contact with the sympathetic neurons. It is concluded that the primitive sympathicoblasts are, in favourable conditions, capable of differentiation in culture up to mature sympathetic neurons.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 39 (1974), S. 129-142 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cholinesterases were demonstrated light and electron microscopically completed with fluorescence microscopic studies in the cultured sympathetic ganglion cells of chick embryos. The sympathetic neurons were cultured for various periods in organotypic cultures with Nerve Growth Factor. The cholinesterase reactions were performed according to the Koelle-Gomori, Karnovsky-Roots and Lewis-Shute modifications. Successive demonstrations of amine fluorescence and cholinesterases were carried out. Variable intensity of enzymatic activity was found in the nuclear and cytoplasmic membranes, in the endoplasmic reticulum and in the Golgi apparatus. Nerve fibers were also surrounded with the reaction product. In the specimens processed according to the Karnovsky-Roots modification, which gave a good ultrastructural preservation, reaction product was observed in addition to the neurons also in the cytoplasmic membranes of the cells containing large dense cored vesicles and in the outer membranes of the fibres containing small dense cored vesicles. It is concluded that the cells populating the sympathetic ganglion of chick embryo maintain acetylcholinesterase activity in the organotypic culture without preganglionic innervation or other exogenous stimulation.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 48 (1976), S. 43-50 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The catecholamine-stimulated adenylate cyclase activity was histochemically demonstrated at ultrastructural level in the cultured neurons of the sympathetic ganglia of the chick embryo. Adenylate cyclase activity was demonstrated in the axolemma of the axons and axon terminals, however not inside the axons. Heavy precipitates of the enzymatic reaction product was observed intracellularly in some cell processes identified as dendrites of the sympathetic neurons. In contrast to the extensive reaction in the neuropil, the reaction for the catecholamine-stimulated adenylate cyclase was scarse in the perikarya of the neurons. Only occasional spots of adenylate cyclase activity could be demonstrated in the plasmalemma of some neurons, and no reaction product was observed in the cytoplasm. No difference in the localization of the adenylate cyclase activity was observed when either dopamine or noradrenaline were used for stimulation. The possible roles of the catecholamine-stimulated adenylate cyclase in the function of the interneuronal connnections between the aminergic neurons of the sympathetic ganglion are discussed.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 40 (1974), S. 371-376 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Lumbar spinal ganglia of the 12 days incubated chick embryo were cultured in organotypic cultures for up to one month. Cholinesterases were demonstrated histochemically using the thiocholine method according to the modification by Lewis and Shute at light and electron microscopical level. The acetylcholinesterase activity: At light microscopical level the neuron perikarya showed variable intensity of the reaction. In the nerve fibres moderate activity was observed with heavy precipitates of the enzymatic reaction product in the tips of the fibres. At electron microscopical level the reaction product was found in the endoplasmic reticulum and in the nuclear envelope. The plasmalemma of the nerve fibres showed enzymatic activity while the cytoplasmic membranes of the perikarya were devoid of the reaction product. The non-specific cholinesterase activity: No reaction product was observed in the cultures at light microscopical level. The results are compared with the earlier in vivo and in vitro studies and the differences in the enzymatic activity of the spinal and sympathetic ganglia are discussed.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-7381
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The sympathetic ganglia of adult and aged humans were obtained during vascular, gynaecological and urological surgery, and studied using the formaldehyde-induced fluorescence (FIF) method for histochemical demonstration of catecholamines. Microspectrofluorimetry was applied to characterize the emission spectra of the fluorophores. The sympathetic ganglia contained two types of cells exhibiting FIF: the adrenergic neurons and the small, intensely fluorescent (SIF) cells. The adrenergic neurons contained pigment granules exhibiting autofluorescence with emission maximum at 530–540 nm. The number of these granules increased with age while the FIF of the neurons decreased. In the oldest patients, most of the neurons in the sympathetic ganglia showed only pigment fluorescence but not FIF. The decrease of the perikaryonal catecholamine stores is interpreted as a degenerative change leading to impaired function of the neurons. The patterns of AChE did not change with age. The SIF cells were sparse and were mostly observed near the ganglia, forming paraganglia of varying size. Because of the rarity of the SIF cells, the effect of ageing could not be judged reliably.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 226 (1982), S. 51-62 
    ISSN: 1432-0878
    Keywords: Neurohypophysis ; Ultrastructure ; Ageing ; Hormone-containing vesicles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The neurohypophyses (neural lobes of the pituitary) of young adult (3–6 months) and aged (12–30 months) male rats were studied by means of electron microscopy. Stereomorphometric analyses were performed to determine the size, number and relative volume of the hormone-containing vesicles. The principal observations included: 1) the conspicuous depletion in aged rats of the granular hormone-containing vesicles from the axon terminals and the Herring bodies, with a decrease in the relative volume fraction of the vesicles from 4.8 Vv % in the control animals to 1.1 Vv % in the aged rats; 2) a change in the size-distribution of the hormone vesicles; 3) an increase in the extracellular space around the nerve terminals, axons and capillaries; and 4) lipid accumulations and signs of activation in the pituicytes. The possible physiological significance of the findings is discussed in the light of several regulatory functions known to be altered during the process of ageing.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-0878
    Keywords: Sympathetic ganglion ; Chick embryo ; Hydrocortisone ; Culture ; Fluorescence microscopy, electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Lumbar sympathetic ganglia of 12-day-old chick embryos were cultured in organ cultures for 14 days with 1, 10 or 100 mg/l of hydrocortisone or without it. Catecholamines were demonstrated by the formaldehyde-induced fluorescence method. For electron microscopy, the cultures were fixed with glutaraldehyde and osmium tetroxide. Two types of cells with catecholamine fluorescence were observed in the control cultures: (1) weakly fluorescent sympathetic neurons and sympathicoblasts with long nerve fibres, which were the most common cell type in the explant, and (2) brightly fluorescent cells with or without fluorescent processes, which were less common and were scattered in the explant. Hydrocortisone caused a great increase in the number of the brightly fluorescent cells. With 10 mg/l of hydrocortisone the increase was about ten-fold as compared with the control cultures. There was no change in the morphology of the cells, nor could any change be observed in the fluorescence intensity by eye. Electron microscopically the mature neurons were the most common cell type on the surface of the culture, while more immature sympathicoblasts were seen in the deeper layers. Cells were also found which contained large numbers of catecholamine-storing granular vesicles 105–275 nm in diameter. These cells were infrequent. They had round vesicular nuclei and resembled also in other respects sympathicoblasts or young nerve cells. One such cell was found in mitotic division by electŕon microscopy. Hydrocortisone caused a marked increase in the number of these granule-containing cells and their processes. Cells which could have been classified as the small intensely fluorescent cells of the mammalian ganglion type or their electron microscopic equivalent, the granule-containing cells were found neither in the control cultures nor in the hydrocortisone-containing cultures. It is concluded that most brightly fluorescent cells in cultured sympathetic ganglia of the chick are nerve cells or sympathicoblasts rich in amine-storing granular vesicles.
    Type of Medium: Electronic Resource
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