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Fig. 6 | BMC Pharmacology and Toxicology

Fig. 6

From: The grass isn’t always greener: The effects of cannabis on embryological development

Fig. 6

Cannabinoid agonists stimulate ERK phosphorylation in rat hippocampal slices. a Rat hippocampal slices were incubated at 35 °C, as described in Materials and Methods, for 50 min before the addition of vehicle (Control), 1 μM anandamide, 1 μM 2-AG, 1 μM CP 55940, 100 μM WIN 55212–2, 0.2 μM LPA, or 0.1 μM Δ9-THC for 5 min, in the absence or in the presence of 100 μM SR 141716A applied 30 min before. Slices were homogenized in SDS; 60 μg of protein per sample were subjected to immunoblot analysis using antibodies specific for the dually phosphorylated (active) forms of ERK1 and ERK2 (Blot P-ERK). After stripping, the membranes were reprobed with anti-ERK (Blot ERK) antibodies. b For quantification the optical densities of P-ERK2-immunoreactive bands were measured, normalized to the optical densities of total ERK2 in the same samples, and expressed as percentages of controls. Data correspond to means ± SEM. Statistical analysis was done with ANOVA (F (13,24) = 22.8; p < 0.0001) followed by t test (treated vs control: ***p < 0.001, **p < 0.01; treated in the presence of SR141716A vs in its absence: °° p < 0.01, ° p < 0.05). c, d Quantification of the effects of 2-AG on ERK2 active form: time course (drug concentration 1 μM) (c); concentration–response curve (treatment for 5 min) (d). Immunoreactivity was quantified by scanning densitometry using NIH image 1.62 software. Values are means ± SEM of four to eight independent experiments and are expressed as percentages of the maximal increase above unstimulated control values [49]. With open permission from the Journal of Neuroscience

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