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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Experimental brain research 63 (1986), S. 321-330 
    ISSN: 1432-1106
    Keywords: Rat neuroblast ; Proliferation ; Maturation ; Meningeal influence ; Culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Neuronal cells were obtained by dissociating cells from the cerebral hemispheres of rat embryos (10 to 17-day-old), either cleaned entirely or only partially of their meningeal membranes. These cells were seeded on poly-lysine-coated Petri dishes in serum-containing medium. The cultures most enriched in neuronal cells were obtained from brains of 13- to 15-day-old embryos and after 2 h, the culture medium was switched to Dulbecco's modified Eagle's medium, without serum, supplemented with the N1 supplements as described by Bottenstein et al. (1980). The proliferation of neuroblasts from 13-day-old embryos in the presence or absence of meningeal cells was studied by using a combination of tritiated thymidine autoradiography and immuno-staining against neurofilament proteins. The neuroblasts seem to proliferate during the first 3 days. The proliferative activity was further enhanced in the presence of meningeal cells. The glioblasts multiply only after a period of one week in culture conditions as observed here. The subsequent development of the neuroblasts was followed over a period of 4 weeks and the ultrastructural appearance of these cells was investigated at 2 and 3 weeks. In the presence of meningeal cells, many neurons, intensely stained for neurofilament proteins, survived for 21 days, while in control cultures they underwent massive degeneration after 2 weeks. Synapses with numerous clear vesicles were abundant in cultures grown under the influence of meningeal cells; they were rare and possessed few vesicles in control cultures. The data indicate that meningeal cells affect the proliferation and maturation of rat neuroblasts in culture.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-6903
    Keywords: Ion fluxes ; growth factor ; astrocytes ; rat ; primary cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Primary cultures of rat astroglial cells were maintained in a serum-free medium. After 8–10 days of cultivation the cells were exposed to an astroglial growth factor (AGF2) for short periods (1–120 min). Subsequently, uptake of22Na+ and42K+ into control and AGF2-pretreated cells was studied. Assay of the Na+ and K+ values in the cells was also performed by atomic absorption spectrometry. Treatment of rat astroglial cells with AGF2 resulted in a significant increase of the uptake of both Na+ and K+ depending on the duration of the exposure period. To reach the maximum increase of cation uptake, 6–10 min and 30 min of AGF2 pretreatment were needed for Na+ and K+, respectively. Amiloride blocked this increase of Na+ and K+ uptake elicited by AGF2 pretreatment, but the control cells were amiloride resistant. Treatment with AGF2 increased the ouabain sensitivity of the K+ uptake as that: 10−4 M ouabain inhibited K+ uptake of the AGF2-treated cells to the same degree as 5×10−3 M ouabain with the control cells. The Na+ uptake of AGF2-treated cells, however, exhibited no relevant changes in the presence of ouabain. A significant part of the AGF2-induced K+ uptake could be inhibited by both ouabain and amiloride, but a ouabain-resistant and amiloride-sensitive component also was revealed. The furosemide sensitivity of both Na+ and K+ uptake into cultured astroglial cells was also significantly increased by AGF2. Our findings suggest that short-term exposure of cultured glial cells to AGF2 induces these very early ionic events: 1) The appearance of a relevant amiloride-sensitive Na+/H+ exchange, and as a consequence of increased Na+ entry into the cells, secondary activation of the ouabain-sensitive K+ uptake via the Na+,K+-pump. 2) A direct effect of AGF2 on the Na+,K+-pump assembly in the membrane, resulting in increased Na+ sensitivity of the inner pump sites and enhanced ouabain sensitivity of the external K+-binding sites. 3) An increase of ouabain-resistant but amiloride- or furosemide-sensitive Na+ and K+ uptake.
    Type of Medium: Electronic Resource
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