Recent data from several laboratories have pointed to several roles of endocannabinoids in embryonic and adult neurogenesis. However, definitive conclusions are still lacking due to conflicting results originating from different cellular models or experimental timeframes. In the present work we employed ST14A cells, a line of immortalised neural progenitors derived from embryonic day 14 rat striatum primordia. Due to their characteristics, these cells represent a valuable tool to investigate cannabinoid functions on neural precursors undergoing proliferation and differentiation. We found that ST14 cells express both cannabinoid receptor 1 (CB1) and 2 (CB2) by RT-PCR and ICC. CB2 expression increases with time in culture, both under proliferating (10% FBS) and differentiating (serum free) conditions. We began to study the effects of CB2 activation by incubating ST14A cells for 48 hrs in increasing concentrations of a CB2-specific agonist (JWH133). We found a decrement in the number of cells with 10 nM JWH133. This decrement was specifically reversed by co-incubation with 1 mM AM630, a specific CB2 antagonist. BrdU incorporation in the presence of JWH133 showed a moderate decrease in the proliferation rate compared to controls, indicating a possible role of CB2 in cell-cycle regulation. Morphological evaluation did not show cell sufferance, suggesting that CB2 stimulation does not induce cell death at the applied drug concentrations. However, we observed a change in cell morphology and in the pattern of staining of cytoskeletal components, such as the intermediate filament nestin. Stimulated cells showed reduced contact area between cells and a tendency to elongation, suggesting a possible involvement of CB2 in cell migration. In summary, in this work we provide preliminary evidence for multiple roles of the cannabinergic system in ST14A progenitors, indicating them as a promising system to characterize cellular and molecular mechanisms of endocannabinoid functions in neurogenesis.
Characterization of the endocannabinoid system in ST14A neural precursors
POMATTO, VALENTINA;COTTONE, Erika;BOVOLIN, Patrizia;FRANZONI, Maria Fosca
2011-01-01
Abstract
Recent data from several laboratories have pointed to several roles of endocannabinoids in embryonic and adult neurogenesis. However, definitive conclusions are still lacking due to conflicting results originating from different cellular models or experimental timeframes. In the present work we employed ST14A cells, a line of immortalised neural progenitors derived from embryonic day 14 rat striatum primordia. Due to their characteristics, these cells represent a valuable tool to investigate cannabinoid functions on neural precursors undergoing proliferation and differentiation. We found that ST14 cells express both cannabinoid receptor 1 (CB1) and 2 (CB2) by RT-PCR and ICC. CB2 expression increases with time in culture, both under proliferating (10% FBS) and differentiating (serum free) conditions. We began to study the effects of CB2 activation by incubating ST14A cells for 48 hrs in increasing concentrations of a CB2-specific agonist (JWH133). We found a decrement in the number of cells with 10 nM JWH133. This decrement was specifically reversed by co-incubation with 1 mM AM630, a specific CB2 antagonist. BrdU incorporation in the presence of JWH133 showed a moderate decrease in the proliferation rate compared to controls, indicating a possible role of CB2 in cell-cycle regulation. Morphological evaluation did not show cell sufferance, suggesting that CB2 stimulation does not induce cell death at the applied drug concentrations. However, we observed a change in cell morphology and in the pattern of staining of cytoskeletal components, such as the intermediate filament nestin. Stimulated cells showed reduced contact area between cells and a tendency to elongation, suggesting a possible involvement of CB2 in cell migration. In summary, in this work we provide preliminary evidence for multiple roles of the cannabinergic system in ST14A progenitors, indicating them as a promising system to characterize cellular and molecular mechanisms of endocannabinoid functions in neurogenesis.
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
