Comparison of the Effects of Ecstasy and Methamphetamine on the RB1 Mouse Embryonic Stem Cells Differentiation into Neural Cells

Background: N-Methyl-3,4-Methylenedioxymethamphetamine (MDMA) and methamphetamine (MA) from the amphetamine family, which are respectively known as ecstasy and ice, have well-established neurotoxic effects on the serotonergic and dopaminergic systems. Regarding the fact that MDMA and MA are primarily used as recreational drugs and are commonly used during child bearing period, there is a major concern on the embryonic and fetal toxicity of these drugs. We have used different methods for screening models but recently embryonic stem cells give us new method for easier and faster way in evaluation of potential toxicity of drug. Methods: We used embryonic stem cells-derived neuronal cells to determine 50% infectious does (ID50) in both groups. ID50C reflects 50% inhibition of embryonic stem cells on neural differentiation. The effect of MDMA and MA on neural differentiation was assessed during two periods: group 1 (throughout the process of neural differentiation), group 2 (during post-plating). Afterwards, the cells were evaluated for neuronal markers by reverse transcription- polymerase chain reaction (RT-PCR). A concentration range of MDMA (0.1, 1, 10, 100, 200, 500, 750, 1000µM) and of MA (10, 50, 70, 100, 200,500, 750, 1000 µM) was applied to the cells from day 0 onwards throughout the entire culture duration as described. Findings: Considering the fact that MDMA and MA are potent neurotoxic drugs, we evaluated their effects during neuronal differentiation and on the survival of embryonic derived neurons. The ID50 for neural differentiation was 50 µM and 130 µM, while for neuronal survival was 120 µM and 400 µM, respectively in MDMA and MA groups. At 7 days post-plating, RT-PCR analysis revealed that unlike the nestin, the expression of MAP2 at concentrations higher than 100 and 200 µM of MDMA and 200 and 500 µM of MA in groups 1 and 2, respectively, was significantly reduced. Conclusion: Our findings have documented that early phase of neural development which coincides with neural tube formation is more sensitive than the later phase when neural precursor cells differentiate into mature neurons. Regarding the embryonic stem cells capabilities such as tissue-specific properties especially in the formation of neuroectodermal and mesodermal cells, they could be served as a system to evaluate inhibiting or inducing effects on differentiation processes in early embryonic stages. Keywords: Stem cells, Cell differentiation, Toxicity, N-Methyl-3,4-methylenedioxyamphetamine, Methamphetamine