Evaluation of 6-Hydroxydopamine and Rotenone In Vitro Neurotoxicity on Differentiated SH-SY5Y Cells Using Applied Computational Statistics

Abstract

With the increase in life expectancy and consequent aging of the world’s population, the prevalence of many neurodegenerative diseases is increasing, without concomitant improvement in diagnostics and therapeutics. These diseases share neuropathological hallmarks, including mitochondrial dysfunction. In fact, as mitochondrial alterations appear prior to neuronal cell death at an early phase of a disease’s onset, the study and modulation of mitochondrial alterations have emerged as promising strategies to predict and prevent neurotoxicity and neuronal cell death before the onset of cell viability alterations. In this work, differentiated SH-SY5Y cells were treated with the mitochondrial-targeted neurotoxicants 6-hydroxydopamine and rotenone. These compounds were used at different concentrations and for different time points to understand the similarities and differences in their mechanisms of action. To accomplish this, data on mitochondrial parameters were acquired and analyzed using unsupervised (hierarchical clustering) and supervised (decision tree) machine learning methods. Both biochemical and computational analyses resulted in an evident distinction between the neurotoxic effects of 6-hydroxydopamine and rotenone, specifically for the highest concentrations of both compounds.

Novel Use of Cell Profiling Technology to Visualize Mitochondrial Responses of Humpback Whale Fibroblasts to Chemical Exposure

“…No clear dose-dependent response was observed for any chemical, with the exception of the assay positive control, rotenone. Interestingly, significant mitochondrial texture modulation of HuWa TERT to rotenone (approximately 17.5%, Figure a) at 4 nM is 10 to 100 times lower than the in vitro mitochondrial morphology and function responses studies previously reported on different terrestrial mammals, such as human HepG2, SH-SY5Y, and rat cortical astrocytes cells. − Comparable findings have also been reported on medical research council cell strain 5, developed from human fetal lung tissue …”