Cellular viability effects of fatty acid amide hydrolase inhibition on cerebellar neurons

Abstract

The endocannabinoid anandamide (ANA) participates in the control of cell death inducing the formation of apoptotic bodies and DNA fragmentation. The aim of this study was to evaluate whether the ANA degrading enzyme, the fatty acid amide hydrolase (FAAH), would induce cellular death. Experiments were performed in cerebellar granule neurons cultured with the FAAH inhibitor, URB597 (25, 50 or 100 nM) as well as endogenous lipids such as oleoylethanolamide (OEA) or palmitoylethanolamide (PEA) and cellular viability was determined by MTT test. Neurons cultured with URB597 (25, 50 or 100 nM) displayed a decrease in cellular viability. In addition, if cultured with OEA (25 nM) or PEA (100 nM), cellular death was found. These results further suggest that URB597, OEA or PEA promote cellular death.

Figure 1

Photomicrograph of cerebellar granule cells incubated only with culture media (control) or treated with URB597 at 10, 25, 50 or 100 nM (Panels A-E, respectively). The cellular viability (Panel F) was determined by MTT and data is presented as mean ± SEM (%). Scale bar, 100 μm (* vs control/vehicle, p < 0.05).

Figure 2

Photomicrograph of cerebellar granule cells incubated only with culture media (control) or treated with OEA at 10, 25, 50 or 100 nM (Panels A-E, respectively), the remaining cells revealed swollen soma and fragmented extensions. The cellular viability (Panel F) was determined by MTT and data is presented as mean ± SEM (%). Scale bar, 100 μm (* vs control/vehicle, p < 0.05).

Figure 3

Photomicrograph of cerebellar granule cells incubated only with culture media (control) or treated with PEA at 10, 25, 50 or 100 nM (Panels A-E, respectively). The cellular viability (Panel F) was determined by MTT and data is presented as mean ± SEM (%). Scale bar, 100 μm (* vs control/vehicle, p < 0.05).