Non-Serotonergic Neurotoxicity by MDMA (Ecstasy) in Neurons Derived from Mouse P19 Embryonal Carcinoma Cells

Abstract

3,4-methylenedioxymethamphetamine (MDMA; ecstasy) is a commonly abused recreational drug that causes neurotoxic effects in both humans and animals. The mechanism behind MDMA-induced neurotoxicity is suggested to be species-dependent and needs to be further investigated on the cellular level. In this study, the effects of MDMA in neuronally differentiated P19 mouse embryonal carcinoma cells have been examined. MDMA produces a concentration-, time- and temperature-dependent toxicity in differentiated P19 neurons, as measured by intracellular MTT reduction and extracellular LDH activity assays. The P19-derived neurons express both the serotonin reuptake transporter (SERT), that is functionally active, and the serotonin metabolizing enzyme monoamine oxidase A (MAO-A). The involvement of these proteins in the MDMA-induced toxicity was investigated by a pharmacological approach. The MAO inhibitors clorgyline and deprenyl, and the SERT inhibitor fluoxetine, per se or in combination, were not able to mimic the toxic effects of MDMA in the P19-derived neurons or block the MDMA-induced cell toxicity. Oxidative stress has been implicated in MDMA-induced neurotoxicity, but pre-treatment with the antioxidants α-tocopherol or N-acetylcysteine did not reveal any protective effects in the P19 neurons. Involvement of mitochondria in the MDMA-induced cytotoxicity was also examined, but MDMA did not alter the mitochondrial membrane potential (ΔΨm) in the P19 neurons. We conclude that MDMA produce a concentration-, time- and temperature-dependent neurotoxicity and our results suggest that the mechanism behind MDMA-induced toxicity in mouse-derived neurons do not involve the serotonergic system, oxidative stress or mitochondrial dysfunction.

In silico peptide-directed ligand design complements experimental peptide-directed binding for protein–protein interaction modulator discovery

“…2C). 40,41 BxPC-3 cells were incubated with compound 1-4 or positive control, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), followed by TMRE. TMRE readily accumulates in the mitochondria resulting in a fluorescent signal.…”

In Silico Peptide-Directed Ligand Design Complements Experimental Peptide-Directed Binding for Protein-Protein Interaction Modulator Discovery

“…A tetramethylrhodamine ethyl ester (TMRE) assay was employed to evaluate if compounds generated mitochondrial membrane depolarization, one of the first signs of Mcl-1 inhibition ( Figure 2C). 38,39 BxPC-3 cells were incubated with compounds 1-4 or positive control, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), followed by TMRE. TMRE readily accumulates in the mitochondria resulting in a fluorescent signal.…”

Protective Effects of Vitamin E on Heart and Testis Histology Following MDMA (Ecstasy) Exposure in Mice

“…Some tissues such as the brain, heart, liver, kidney, and testis may also be damaged, leading to dangerous consequences depending on the damage intensity (2). A general damage of these factors is production of oxidative stress that plays the main role in MDMA pathogenesis (3,4). By increasing the heart rate and blood pressure, MDMA can potentially stimulate cardiovascular action (5).…”