Biomedical subjects

Francesca Silvagno

Publications and source records attributed to Francesca Silvagno.

3 recordsLinked to original sources

PubMed2003

Synaptic activity induces signalling to CREB without increasing global levels of cAMP in hippocampal neurons.

Nuclear calcium signals associated with electrical activation of neurons can control the activity of the transcription factor cAMP-response element binding protein (CREB). Yet, cAMP is thought to be the key messenger that links synaptic activity to the regulation of CREB-mediated transcription. It is generally assumed that synaptic activity increases the intracellular levels of cAMP; this causes activation of the cAMP-dependent protein kinase (PKA) that regulates CREB-mediated transcription either directly or through controlling nuclear signalling of the MAP kinases/extracellular signal-regulated kinases (ERK1/2) pathway. Here we show that, in hippocampal neurons, synaptic activity failed to increase global levels of cAMP that would be required for the cAMP-PKA system to induce nuclear events. Even near-continuous bursting of action potentials, giving rise to large nuclear calcium signals and robust CREB-dependent transcription, left global intracellular levels of cAMP unchanged. These results suggest that the cAMP-PKA system does not function as the transducer of synaptic signals to the nucleus. They indicate that the known inhibitory effects of blockers of PKA on gene expression and long-lasting plasticity triggered by calcium entry reflect a gating function of basal activity of PKA that renders neurons permissive for nuclear calcium-regulated, CREB/CBP-dependent gene expression.

Action Potentials↗

PubMed2003

Infectious and inflammatory stimuli decrease endothelial nitric oxide synthase activity in vitro.

BACKGROUND: Perturbation of iron metabolism, especially the increase of serum ferritin levels, is often associated with both inflammation and hypertension. Changes in iron availability can affect an important regulator of vascular tone, the endothelial nitric oxide synthase (eNOS), activated by a heme-dependent dimerization. OBJECTIVE: To study the regulation of the anti-hypertensive eNOS in human endothelial cells, in correlation with iron metabolism alterations and stimuli triggering them in vivo, such as inflammation or infection. DESIGN: Cells were treated with stimuli mimicking infection or inflammation [lipopolysaccharide (LPS) and/or tumor necrosis factor alpha (TNFalpha)]. and iron shortage (succinylacetone and desferrioxamine). The effect on eNOS expression and activation was evaluated, as well as ferritin content. METHODS: eNOS protein expression was evaluated by separating the monomeric from the active dimeric form by low-temperature sodium dodecyl sulphate poly-acrylamide gel electrophoresis (SDS-PAGE), and mRNA was analyzed by semi-quantitative reverse transcriptase (RT)-polymerase chain reaction (PCR). As for LPS and TNFalpha, eNOS monomer decreased already after a 72-h treatment and further at 144 h, whereas the down-regulation of the dimer was slower, peaking at 144 h. Succinylacetone and desferrioxamine were effective only at 144 h. The mRNA levels were increasingly reduced after incubation, more markedly by LPS and TNFalpha together, whereas succinylacetone and desferrioxamine had no effect on transcription. We found that endothelial cells are not the source of increased ferritin production. CONCLUSIONS: The results of this study suggest a down-regulating effect of infectious and inflammatory stimuli on eNOS expression, both at the mRNA level and protein expression or stability and dimerization, enhanced by heme and iron shortage, and indicate eNOS as a possible link between infection and hypertension.

Bacterial Infections↗

PubMed2002

Synergistic effect of retinoic acid and dehydroepiandrosterone on differentiation of human neuroblastoma cells.

Retinoic acid (RA) affects many cell types by either promoting their survival or inducing their differentiation. Dehydroepiandrosterone (DHEA), a precursor for both androgenic and estrogenic steroids and abundantly produced by brain, is known as an inhibitor of cell proliferation. Differentiation of a human neuroblastoma cell line (SK-N-BE) was evaluated measuring growth rate, motility, neurite extension and GAP-43 expression. We report that DHEA enhances the differentiating effect of RA on neuroblastoma cells via a signalling that is not RA receptor-mediated. Instead, we show a differential expression of matrix metalloproteinases: RA enhances the activity of MMP-2, whereas MMP-9 expression is up-regulated by DHEA. The concerted modulation of these proteinases may support the neurite outgrowth observed after co-treatment with the two drugs.

Cell Differentiation↗