We induced apoptosis in primary cultures of cerebellar granule neurons by switching the growing medium into a medium containing lower concentrations of K+(5 or 10 mM instead of 25 mM) or, alternatively, by addition of staurosporine. The apoptotic phenotype was always preceded by an early increase in the intracellular levels of the disialoganglioside GD3, which peaked at 2-6 h and returned back to normal at 12 h. GD3 synthase, the enzyme that forms GD3 from the monosialoganglioside GM3, was also induced at early times after the induction of apoptosis in granule cells. Immunofluorescent staining showed that GD3 increased in neuronal cell bodies and neurites, but was never localized in cell nuclei. In cultures switched into a low K+-containing medium, exogenously applied GD3, but not the disialoganglioside GD1a, accelerated the development of neuronal apoptosis. In contrast, the antisense-induced knock-down of GD3 synthase was protective against granule cell death induced by lowering extracellular K+from 25 to 10 - but not 5 - mM. These results demonstrate that an early and transient increase in GD3 synthesis is one of the factors that contribute to the induction of neuronal apoptosis in culture.

Melchiorri, D., Martini, F., Lococo, E., Gradini, R., Barletta, E., De Maria Marchiano, R., Caricasole, A., Nicoletti, F., Lenti, L., An early increase in the disialoganglioside GD3 contributes to the development of neuronal apoptosis in culture, <<CELL DEATH AND DIFFERENTIATION>>, 2002; 9 (6): 609-615. [doi:10.1038/sj.cdd.4401020] [http://hdl.handle.net/10807/113862]

An early increase in the disialoganglioside GD3 contributes to the development of neuronal apoptosis in culture

De Maria Marchiano, Ruggero;
2002

Abstract

We induced apoptosis in primary cultures of cerebellar granule neurons by switching the growing medium into a medium containing lower concentrations of K+(5 or 10 mM instead of 25 mM) or, alternatively, by addition of staurosporine. The apoptotic phenotype was always preceded by an early increase in the intracellular levels of the disialoganglioside GD3, which peaked at 2-6 h and returned back to normal at 12 h. GD3 synthase, the enzyme that forms GD3 from the monosialoganglioside GM3, was also induced at early times after the induction of apoptosis in granule cells. Immunofluorescent staining showed that GD3 increased in neuronal cell bodies and neurites, but was never localized in cell nuclei. In cultures switched into a low K+-containing medium, exogenously applied GD3, but not the disialoganglioside GD1a, accelerated the development of neuronal apoptosis. In contrast, the antisense-induced knock-down of GD3 synthase was protective against granule cell death induced by lowering extracellular K+from 25 to 10 - but not 5 - mM. These results demonstrate that an early and transient increase in GD3 synthesis is one of the factors that contribute to the induction of neuronal apoptosis in culture.
2002
Inglese
Melchiorri, D., Martini, F., Lococo, E., Gradini, R., Barletta, E., De Maria Marchiano, R., Caricasole, A., Nicoletti, F., Lenti, L., An early increase in the disialoganglioside GD3 contributes to the development of neuronal apoptosis in culture, <<CELL DEATH AND DIFFERENTIATION>>, 2002; 9 (6): 609-615. [doi:10.1038/sj.cdd.4401020] [http://hdl.handle.net/10807/113862]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10807/113862
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