Figure 1

The CB₁ receptor protects cultured striatal cells from NMDA-induced excitoxicity via PI3K/Akt/mTORC1/BDNF. (a) STHdh^Q7/Q7 cells were incubated for the times indicated with vehicle, 0.5-μM THC or 10-nM HU-210. Cells were lysed and western blot analyses were conducted. Quantification of mean optical density (O.D.) values relative to those of loading controls (respective total proteins, or α-tubulin in the case of PKA phosphorylated substrates) as well as representative blots are shown (n=3–4 experiments). (b, c) STHdh^Q7/Q7 cells were preincubated for 5 h in Locke’s solution with or without 1-mM NMDA together with vehicle, 0.5-μM THC, 10-nM HU-210, 0.2-μM wortmanin, 0.1-μM Akti-1/2, 30-nM rapamycin and/or 25-nM K252a, and subsequently incubated for 24 h in NMDA-free medium. Relative cell viability is shown (n=6–8 experiments). (d) STHdh^Q7/Q7 cells were transfected with a non-targeted siRNA or with siRNAs directed against BDNF or TrkB, and subsequently incubated for 5 h with or without NMDA, THC and/or HU-210 as in b. Relative cell viability is shown (n=4–6 experiments). (e) Primary mouse striatal neurons were incubated for 30 min in Locke’s medium with or without 50-μM NMDA, together with vehicle, 0.3-μM THC, 10-nM HU-210, 0.25-μM SR141716, 0.1-μM Akti-1/2, 20-nM rapamycin and/or 10-nM K252a, and subsequently incubated for 2 h in NMDA-free medium. Relative cell viability is shown (n=4–6 experiments). Data were analyzed using ANOVA with post hoc Student-Newman-Keuls test. *P<0.05 and **P<0.01 from the corresponding vehicle-treated cells