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Ajay Ashok, Wai Lydia Tai, Anton Lennikov, Kin-Sang Cho, Tor Utheim, Dongfeng Chen; Epigenetic modulation following electrical stimulation in neurons. Invest. Ophthalmol. Vis. Sci. 2022;63(7):2969 – F0210.
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© ARVO (1962-2015); The Authors (2016-present)
Epigenetic factors are known to influence neural development, functionality, and their response to pathophysiology. In recent times, electrical stimulation (ES) of cell and animal models have shown promising results in neural regeneration and recovery. However, the underlying mechanisms remains elusive. Our study will investigate if ES holds the promise to regulate DNA demethylation, a vital epigenetic event known to influence neuronal regeneration, using an in-vitro cell model.
PC12 cells acquired from ATCC were cultured and the effect of ES on DNA demethylases (ten-eleven translocation (TETs) proteins (TET1, TET2 and TET3)) was studied using qPCR. PC12 cells were incubated with 0, 5 and 100 ng/ml of nerve growth factor (NGF), a known inducer of neuronal differentiation, for 5 days and were also electrically stimulated for 20 min at 100 µA. Non-electrically stimulated cells were maintained as control. RNAs were collected 16 h post procedure and the changes in TETs gene expression were analyzed using qPCR. Additionally, lactate dehydrogenase (LDH) estimation assay was carried out to evaluate cytotoxicity in the experimental groups. Two-tailed Student's t-test was used for statistical analysis.
qPCR revealed increased TET1 gene expression in cells exposed to ES relative to control cells. TET2 and TET3 amplification did not change significantly. LDH assay demonstrated no significant cytotoxicity in any treatment groups. Interestingly, increasing NGF concentration in PC12 cells exhibited similar induction of TET1 but not TET2 or TET3 expression, suggesting a possibly shared epigenetic event in ES and NGF-induced neuronal differentiation process.
Our results point to a possible regulation of TET1, a DNA demethylase, by ES in neurons. A comprehensive understanding of epigenetic regulation following ES in neuronal differentiation, development and homeostasis will help us unravel novel molecular pathways and pave the way to design blueprints for effective therapeutics to address neuronal protection, repair, and regeneration.
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.
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