Effects of aconitine on [Ca²⁺] oscillation in cultured myocytes of neonatal rats

Abstract

In order to investigate the effects of aconitine on [Ca²⁺] oscillation patterns in cultured myocytes of neonatal rats, fluorescent Ca²⁺ indicator Fluo-4 NW and laser scanning confocal microscope (LSCM) were used to detect the real-time changes of [Ca²⁺] oscillation patterns in the cultured myocytes before and after aconitine (1.0 μmol/L) incubation or antiarrhythmic peptide (AAP) and aconitine co-incubation. The results showed under control conditions, [Ca²⁺] oscillations were irregular but relatively stable, occasionally accompanied by small calcium sparks. After incubation of the cultures with aconitine, high frequency [Ca²⁺] oscillations emerged in both nuclear and cytoplasmic regions, whereas typical calcium sparks disappeared and the average [Ca²⁺] in the cytoplasm of the cardiomyocyte did not change significantly. In AAP-treated cultures, intracellular [Ca²⁺] oscillation also changed, with periodic frequency, increased amplitudes and prolonged duration of calcium sparks. These patterns were not altered significantly by subsequent aconitine incubation. The basal value of [Ca²⁺] in nuclear region was higher than that in the cytoplasmic region. In the presence or absence of drugs, the [Ca²⁺] oscillated synchronously in both the nuclear and cytoplasmic regions of the same cardiomyocyte. It was concluded that although oscillating strenuously at high frequency, the average [Ca²⁺] in the cytoplasm of cardiomyocyte did not change significantly after aconitine incubation, compared to the controls. The observations indicate that aconitine induces the changes in [Ca²⁺] oscillation frequency other than the Ca²⁺ overload.

Keywords

forensic toxicology / aconitine / [Ca²⁺] oscillation / primary cell culture