Impaired Bioenergetics in Mutant Mitochondrial DNA Determines Cell Fate During Seizure-Like Activity

Abstract

Mutations in genes affecting mitochondrial proteins are increasingly recognised in patients with epilepsy, but the factors determining cell fate during seizure activity in these mutations remain unknown. Fluorescent dye imaging techniques were applied to fibroblast cell lines from patients suffering from common mitochondrial mutations and to age-matched controls. Using live cell imaging techniques in fibroblasts, we show that fibroblasts with mutations in the mitochondrial genome had reduced mitochondrial membrane potential and NADH pools and higher redox indices, indicative of respiratory chain dysfunction. Increasing concentrations of ferutinin, a Ca²⁺ ionophore, led to oscillatory Ca²⁺ signals in fibroblasts resembling dynamic Ca²⁺ changes that occur during seizure-like activity. Co-monitoring of mitochondrial membrane potential (ΔΨ_m) changes induced by ferutinin showed accelerated membrane depolarisation and cell collapse in fibroblasts with mutations in the mitochondrial genome when compared to controls. Ca²⁺ flash photolysis using caged Ca²⁺ confirmed impaired Ca²⁺ handling in fibroblasts with mitochondrial mutations. Findings indicate that intracellular Ca²⁺ levels cannot be compensated during periods of hyperexcitability, leading to Ca²⁺ overload and subsequent cell death in mitochondrial diseases.

Keywords: Ca2+; Epilepsy; Fibroblasts; Mitochondrial mutation; PTP opening.