We confirmed endogenous 2 AG to become active in the complex process of oligodendrocyte differentiation, also demonstrating that oligodendroglial cells express DAGLa, DAGLb natural product librariesand monoacylglycerol lipase, two enzymes responsible for the synthesis and degradation of 2 AG. Oligodendrocyte progenitor differentiation is impaired by the inhibition of DAGL activity with specific pharmacological inhibitors, or disruption of 2 AG synthesis with specific siRNAs against DAG lipases, clearly indicating that 2 AG is important for oligodendrocyte maturation. Here, we confirm and expand on these previous studies showing the importance of basal cannabinoid activity on the differentiation of oligodendrocytes. Indeed, we now show that the service of CB1 or CB2 Papillary thyroid cancer receptors by selective exogenous agonists accelerates oligodendrocyte difference via the PI3K/Akt and mammalian target of rapamycin signalling pathways. Techniques Purification and culture of oligodendrocyte progenitor cells All animal care and experimental procedures complied with Eu legislation and current Spanish. Main blended glial cultures were prepared as described previously and in line with the modified manner of McCarthy and de Vellis. Shortly, the forebrain of new-born Wistar rats was dissociated in 0. 25 percent trypsin by trituration. The cell suspension was filtered through the filtrate centrifuged at 190 and a 150 mmnylon mesh? g for 10 min. The cells were then re-suspended in Dulbeccos modified Eagle medium containing one hundred thousand FCS and plated on poly Lornithine sprayed 75 cm2 flasks. After 10 days in culture, the flasks were shaken at 225 rpm at 37 C for 2 h to remove the loosely adherent microglia, and the remaining OPCs present on top of the confluent monolayer of astrocytes were dislodged by Canagliflozin dissolve solubility shaking overnight at 260 r. G. m. The cell suspension was filtered through a 30 mm nylon mesh and then pre coated on bacterial grade Petri dishes for just two h. The non adherent OPCs that remained in suspension were restored and further purified by immunopanning. Shortly, two 100 mm Petri dishes were incubated over night at 4 C in 10 mL Tris containing affinity purified goat anti mouse IgM. A day later, each dish was washed three times with PBS, and 10 mL of the primary A2B5 antibody was added for 1 h at room temperature. Following a further three washes with PBS, 10 mL of DMEM plus 10% goat serum was put into block non specific binding to the dishes, and it was removed just before the addition of the cell suspension. Cells were put into the plates and after 1 h at room temperature, and the plates were washed again and again with Hanks balanced salt solution. Finally, the adherent cells were released by incubating them in a 0. 125,000-square trypsin solution and then by hand pipetting DMEM plus 10 percent FCS onto the surface of the dish.