Adult neurogenesis occurs in the subventricular zone (SVZ) and in the dentate gyrus of the hippocampus. Once isolated, cells from the SVZ can grow in vitro as clonal clusters of proliferating cells, called neurospheres (NS), which can either self-renew or differentiate to astrocytes, neurons and oligodendrocytes1. There is increasing evidence that nucleotides can play a role in controlling both adult and embryonic neurogenesis2. The aim of our study was to investigate the role of purinergic signaling in controlling the proliferation, the self-renewal capability and the differentiative potential of neural stem cells derived from adult murine SVZ by the NS assay. To this aim, we evaluated if and how the P2Y1,11,12,13 non-selective agonist ADPβS can affect the proliferation and the self-renewal capability of NS. When analyzing the ability of cells deriving from primary NS to constitute new NS, we observed that ADPβS application increased the number of secondary NS, while it had no effect on the total number of viable cells and on the NS size. When the cells derived from the dissociation of secondary NS were used, ADPβS increased the number of tertiary NS whereas a reduction of the total number of viable cells and of the size of the NS was obtained. Since progenitor cells have limited proliferative and self-renewal capacity, and give rise to small-sized neurospheres3, it can be envisaged that ADPβS acts on this specific cell population and promotes the formation of a higher number of NS with a smaller size. We also tested if the exposure to ADPβS changed the differentiative potential of 7-day-old dissociated primary NS, cultured as adherent cells for 7 additional days in presence of 2% FBS. No differences in the proportion among the various cell types derived from neurospheres between treated and untreated cultures were observed: the majority of the cells resulted to be GFAP-positive astrocytes, while few βIII-tubulin-positive neurons and O4-positive oligodendrocytes were detected. We are now evaluating the effect of ADPβS on the differentiative potential of 7-days-old secondary NS plated without dissociation and grown for 7 additional days without FBS and GFs. Preliminary results obtained by a densitometric analysis, suggest an increased presence of both βIII-tubulin+ neuron and GFAP+ astrocytes in ADPβS-treated cultures. References 1. Doetsch F, Caille I, Lim DA, Garcia-Verdugo JM, Alvarez-Buylla A (1999) Subventricular zone astrocytes are neural stem cells in the adult mammalian brain. Cell, 97:703-716. 2. Neary JT and Zimmermann H (2009) Trophic functions of nucleotides in the central nervous system. Trends Neurosci, 32(4):189-98. 3. Gritti A, Bonfanti L, Doetsch F, Caille I, Alvarez-Buylla A, Lim DA, Galli R, Verdugo JM, Herrera DG, Vescovi AL (2002) Multipotent neural stem cells reside into the rostral extension and olfactory bulb of adult rodents. J Neurosci, 22(2):437-45.
|Titolo:||Adult neurogenesis in the subventricular zone: studies on the role of purinergic signalling by the neurosphere assay|
BOCCAZZI, MARTA (Primo)
|Data di pubblicazione:||2011|
|Settore Scientifico Disciplinare:||Settore BIO/14 - Farmacologia|
|Citazione:||Adult neurogenesis in the subventricular zone: studies on the role of purinergic signalling by the neurosphere assay / M. Boccazzi, C. Rubino, C. Rolando, A. Buffo, M. Abbracchio, S. Ceruti. ((Intervento presentato al convegno Next Step2: la giovane ricerca avanza tenutosi a Milano nel 2011.|
|Appare nelle tipologie:||14 - Intervento a convegno non pubblicato|