An elevated consumption of cocaine (benzoylmethylecgonine), which causes anesthetic and
stimulant effects on the central nervous system, may be associated with several
neurodegenerative conditions affecting dopaminergic neurons, such as Parkinson's disease (PD).
To investigate the impact of cocaine on cell viability and morphology, dopaminergic neurons from
the substantia nigra (SN4741) were cultured. Analysis involved assessing cell death (LDH levels)
and cell morphology (GIEMSA staining) after a 24-hour treatment period. Additionally, the
effects on reactive oxygen species (ROS) generation (DH2), membrane potential (JC-1), oxygen
consumption rate (OCR), and mitochondrial stress (Seahorse) were evaluated after a 6-hour
treatment. The optimal concentration of cocaine for experimental use (2 mM) was identified,
inducing a substantial 39.75% neuronal death. Examination of neuronal death (LDH) revealed a
remarkable 280% increase following cocaine treatment. Optical analysis demonstrated
heightened mortality and detrimental changes in neuronal morphology post-cocaine treatment,
including a globose shape, loss of synapses, extremely thin membrane, and cell aggregation. In
the "short time" experiments, mitochondrial oxidative damage was evident in SN cells treated
with cocaine, leading to the demise of 75% of the cells. Furthermore, a significant 173.6%
increase in reactive oxygen species (ROS) production and a 20% reduction in mitochondrial
membrane potential (JC-1 assay) were observed. Cocaine treatment also resulted in a notable
60% decrease in mitochondrial oxygen consumption. In summary, a concentration of 2 mM
cocaine induces a considerable rise in mitochondrial oxidative damage, subsequently causing
morphological alterations and progressive death of dopaminergic neurons due to the
accumulation of reactive oxygen species (ROS).
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