Human Ntera2/cl.D1 (NT2) cells treated with retinoic acid (RA) differentiate towards a well characterized neuronal phenotype sharing many features with human fetal neurons. In view of the emerging role of caspases in murine stem cell/neural precursor differentiation, caspases activity was evaluated during RA differentiation. Caspase-2, -3 and -9 activity was transiently and selectively increased in differentiating and non-apoptotic NT2-cells. SiRNA-mediated selective silencing of either caspase-2 (si-Casp2) or -9 (si-Casp9) was implemented in order to dissect the role of distinct caspases. The RA-induced expression of neuronal markers, i.e. neural cell adhesion molecule (NCAM), microtubule associated protein-2 (MAP2) and tyrosine hydroxylase (TH) mRNAs and proteins, was decreased in si-Casp9, but markedly increased in si-Casp2 cells. During RA-induced NT2 differentiation, the class III histone deacetylase Sirt1, a putative caspase substrate implicated in the regulation of the proneural bHLH MASH1 gene expression, was cleaved to a ∼100 kDa fragment. Sirt1 cleavage was markedly reduced in si-Casp9 cells, even though caspase-3 was normally activated, but was not affected (still cleaved) in si-Casp2 cells, despite a marked reduction of caspase-3 activity. The expression of MASH1 mRNA was higher and occurred earlier in si-Casp2 cells, while was reduced at early time points during differentiation in si-Casp9 cells. Thus, caspase-2 and -9 may perform opposite functions during RA-induced NT2 neuronal differentiation. While caspase-9 activation is relevant for proper neuronal differentiation, likely through the fine tuning of Sirt1 function, caspase-2 activation appears to hinder the RA-induced neuronal differentiation of NT2 cells.
Pistritto, G., Papaleo, V., Sanchez, P., Ceci, C., & Barbaccia, M.L. (2012). Divergent modulation of neuronal differentiation by caspase-2 and -9. PLOS ONE, e36002.
|Tipologia:||Articolo su rivista|
|Citazione:||Pistritto, G., Papaleo, V., Sanchez, P., Ceci, C., & Barbaccia, M.L. (2012). Divergent modulation of neuronal differentiation by caspase-2 and -9. PLOS ONE, e36002.|
|IF:||Con Impact Factor ISI|
|Settore Scientifico Disciplinare:||Settore BIO/14|
|Revisione (peer review):||Esperti anonimi|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1371/journal.pone.0036002.|
|Stato di pubblicazione:||Pubblicato|
|Data di pubblicazione:||mag-2012|
|Titolo:||Divergent modulation of neuronal differentiation by caspase-2 and -9.|
|Autori:||Pistritto, G; Papaleo, V; Sanchez, P; Ceci, C; Barbaccia, ML|
|Appare nelle tipologie:||01 - Articolo su rivista|