Cardiac Myosin and Thin Filament as a Target for Lead and Cadmium Divalent Cations

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Abstract

Lead and cadmium, which are heavy metals widely distributed in the environment, significantly contribute to cardiovascular morbidity and mortality. Using Leadmium Green dye, we have shown that lead and cadmium enter the cardiomyocytes, distributing throughout the cell. Using an in vitro motility assay, we have shown that the sliding velocity of actin and native thin filaments over myosin decreases with increasing concentrations of Pb2+ and Cd2+. Significantly lower concentrations of Pb2+ and Cd2+ (0.6 mM) were required to stop the movement of thin filaments over myosin compared to stopping actin movement over the same myosin (1.1-1.6 mM). A lower concentration of Cd2+ (1.1 mM) needed to stop actin movement over myosin compared to the Pb2++Cd2+ combination (1.3 mM) and lead alone (1.6 mM). There were no differences found in the lead and cadmium cations’ effects on the relative force developed by myosin heads or the number of actin filaments bound to myosin. The sliding velocity of actin over myosin in the left atrium, right and left ventricles changed equally when exposed to the same dose of the same metal. Thus, we have demonstrated for the first time that Pb2+ and Cd2+ can directly affect myosin and thin filament function, with Cd2+ exerting a more toxic influence on myosin function compared to Pb2+.

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O. P. Gerzen

Institute of Immunology and Physiology of the Ural Branch of the Russian Academy of Sciences

Author for correspondence.
Email: o.p.gerzen@gmail.com
Russian Federation, Ekaterinburg

Yu. K. Potoskueva

Institute of Immunology and Physiology of the Ural Branch of the Russian Academy of Sciences

Email: o.p.gerzen@gmail.com
Russian Federation, Ekaterinburg

A. E. Tzybina

Institute of Immunology and Physiology of the Ural Branch of the Russian Academy of Sciences

Email: o.p.gerzen@gmail.com
Russian Federation, Ekaterinburg

T. A. Myachina

Institute of Immunology and Physiology of the Ural Branch of the Russian Academy of Sciences

Email: o.p.gerzen@gmail.com
Russian Federation, Ekaterinburg

L. V. Nikitina

Institute of Immunology and Physiology of the Ural Branch of the Russian Academy of Sciences

Email: o.p.gerzen@gmail.com
Russian Federation, Ekaterinburg

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2. Fig. 1. Rat left ventricular cardiomyocyte stained with Leadmium Green in light-field mode (a), at the 0th (b) and 10th minute (c) of PbCl2 treatment in fluorescence mode; normalised fluorescence intensity at the 0th and 10th minute of PbCl2 treatment (d), data are presented as mean ± standard deviation. Rat left ventricular cardiomyocyte stained with Leadmium Green in light-field mode (e), at the 0th (f) and 10th minute (g) of CdCl2 treatment in fluorescence mode; normalised fluorescence intensity at the 0th and 10th minute of CdCl2 treatment (h), data are presented as mean ± standard deviation

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3. Fig. 2. Dependence of actin sliding velocity on myosin on the concentration of PbCl2 (a), CdCl2 (b) and combination of PbCl2 and CdCl2 (1 : 1) (c); dependence of mobile filament fraction on the concentration of PbCl2 (d), CdCl2 (e) and combination of PbCl2 and CdCl2 (1 : 1) (f)

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4. Fig. 3. Dependence of actin myosin sliding velocity on the fraction of NEM-modified myosin in the control group (K), upon addition of 0.5 mM PbCl2 (a), 0.5 mM CdCl2 (b) and 0.5 mM combination of PbCl2 and CdCl2 (1 : 1) (c). Number of filaments in the field of view of the experimental recording when PbCl2 (d), CdCl2 (e) and the combination of PbCl2 and CdCl2 (1 : 1) (f) were added

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5. Fig. 4. Normalised actin sliding velocity on myosin of pig atria (a), right (b) and left (c) ventricle in the presence of 0.1 mM and 0.5 mM PbCl2 or CdCl2. * Statistically significant differences from the control group (p < 0.05)

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6. Fig. 5. Normalised sliding velocity of pig left ventricular actin (a) and NTP (b) on pig left ventricular myosin in the presence of 0.1 mM and 0.5 mM PbCl2, CdCl2 or their combination. * Statistically significant differences from the control group (p < 0.05)

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