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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Journal of neuroendocrinology 9 (1997), S. 0 
    ISSN: 1365-2826
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Both pancreatic beta cells (insulin-secreting cells) and neuronal cells express functional receptors for nerve growth factor. However, while the effect of nerve growth factor on neuronal differentiation is well known, its role on pancreatic beta cells is not established. It has been demonstrated that in PC12 cells, a well characterized NGF-responsive cell line, NGF increases the production of nitric oxide by inducing the expression of nitric oxide synthase. Nitric oxide is subsequently responsible for growth arrest, a step necessary for neuronal differentiation, visualized by the extension of neuronal-like processes. In the present study, we studied the effect of nerve growth factor on nitric oxide synthesis in INS-1 cells, an insulin-producing cell line which possesses the machinery necessary to respond to nerve growth factor. It was demonstrated that the expression of none of the three isoforms of nitric oxide was induced by nerve growth factor in INS-1 cells, strongly suggesting that nerve growth factor does not induce an increase in nitric oxide production in this cell line. Finally, we demonstrated that whereas growth arrest occurred in INS-1 cells cultured in the presence of a donor of nitric oxide (SNP), the simultaneous addition of SNP and nerve growth factor is not sufficient to induce the extension of neuronal-like processes in INS-1 cells. These dissimilarities strongly suggest that NGF plays a different role in neuronal and pancreatic beta cells.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1365-2826
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Like neuronal cells, insulin producing cells (beta cells) possess nerve growth factor (NGF) binding sites and express mRNA coding for the low- and high-affinity NGF receptors, p75NGFR and Trk-A respectively. Although the role of NGF on neuronal cells is well documented, its function on beta cells is still unknown. As a first step towards the elucidation of the role of NGF on beta cells, we have characterized both types of NGF receptors on INS-1 cells, a beta cell line derived from a rat insulinorna and studied some early post-receptor events by comparing the signaling pathway of NGF in those cells and in PC12 cells, a well characterized NGF-responsive cell line. By polymerase chain reaction, immunocytochemistry, cross-linking and Western blot analysis, we clearly demonstrated that Trk-A and p75NGFR, the two NGF receptors expressed in INS-1 cells and PC12 cells are similar. Moreover, upon NGF treatment, Trk-A is phosphorylated on tyrosine residues in both cell types in the same dose- and time-dependent manner. These data clearly demonstrate that the first step of NGF signal transduction is similar in PC12 and INS-1 cells. Although early responsive genes like NGFI-A and c-fos are induced in both cell types upon NGF treatment, the induction of c-jun expression is restricted to PC12 cells. Furthermore, the expression of late responsive genes, such as vgf and transin, which are induced by NGF in PC12 cells, are not induced in INS-1 cells. In conclusion, although the initial steps of NGF signal transduction are similar in PC12 and INS-1 cells, some of the later differ. These dissimilarities could suggest that NGF plays different roles in neuronal and pancreatic beta cells.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of neuroendocrinology 7 (1995), S. 0 
    ISSN: 1365-2826
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Insulin producing cells of the pancreas (beta cells) and neuronal cells share a large number of similarities. For example, different molecules, thought to be specific of neuronal cells, are expressed by beta cells. The factors regulating the expression of these molecules in beta cells are poorly understood. In the present work, we have studied the effect of dexamethasone, a synthetic glucocorticoid, on the expression of three different neuronal traits expressed by INS-1 cells, a highly differentiated beta cell line. We demonstrate that dexamethasone treatment decreases the steady state levels of mRNAs coding for both the low-and the high-affinity NGF receptors and of mRNA coding for NF-H, an intermediate neurofilament specific of neurons. This effect was time-dependent, the decrease being detectable after 4–8 h treatment. The decrease in NGF receptors mRNAs steady state levels was paralleled by a decrease in the number of NGF binding sites as demonstrated after Scatchard analysis. We further focused on the mechanisms by which dexamethasone affects the expression of the low affinity NGF receptor. The effect is countered by the glucocorticoid antagonist RU486, indicating that it is mediated by the glucocorticoid receptor. Finally, the decrease in the low-affinity nerve growth factor receptor mRNA steady state level after dexamethasone treatment is not due to mRNA destabilization but can be rather explained through a change in gene transcription.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2013
    Keywords: Key words: pancreatic development ; TGF-β ; differential mRNA display ; glucagon ; B-carboxypeptidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract. The effect of TGF-β on gene activation in embryonic pancreatic rudiments was investigated using differential mRNA display. Several cDNA bands were augmented and some were suppressed in the presence of TGF-β. Differentially expressed cDNAs were re-amplified, sequenced, and sequences compared to the GeneBank database. Glucagone and brain α-tropomyosin cDNAs were identified from the group of augmented cDNAs, and B-carboxypeptidase form the group of suppressed cDNAs. PCR experiments were confirmed with Northern blots. Obtained results are in accordance with immunohistochemical findings and render differential mRNA display a useful technique in identifying differentially expressed genes in embryonic pancreatic rudiments. Several unknown differentially expressed cDNA sequences obtained in our experiments remain to be identified.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Grafts of various types of cells have been performed using bioresorbable polymer matrices. These synthetic fibers are degraded by hydrolysis into normal metabolic intermediates and induce a number of events that are conducive to healing and/or repair, the most important of which may be angiogenesis. The use of biodegradable meshes to deliver genetically altered cells was studied. A β-galactosidase gene was inserted into Long-Evans rat bone marrow stromal (BMS) cells or fibroblasts derived from C57BL/6J mouse embryos using the retroviral vector LNL-SLXβgal. Expression was monitored using X-gal staining. X-gal+ cells from monolayer cultures were seeded onto either polyglycolic acid (PGA) or polyglactin (PGL) biodegradable meshes and grown to confluence. Two types of grafts were performed: (1) embryonic C57BL/6J mouse fibroblasts (EMF) into either nude mice or adult C57BL/6J mice, and (2) Long-Evans rat BMS into Long-Evans rats. β-Galactosidase activity was found for up to 152 days for EMF in nude mice, 123 days for EMF in adult C57BL/6J mice, and 90 days for grafts of syngeneic BMS cells into Long-Evans rats. Noninfected cells grafted using the same methods did not stain with X-gal.
    Type of Medium: Electronic Resource
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