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Decreased expression of sarcoplasmic reticulum (SR) Ca2+-ATPase (SERCA), overexpression of Na+/Ca2+ exchanger (NCX) and diastolic SR Ca2+ leak from the ryanodine receptors (RyRs) are believed to contribute to the decrease of myocyte contraction in failing heart. In this work we induced Ca2+ leak through RyRs in isolated myocytes of guinea pig hearts by 20µM FK-506. The SR Ca2+ leak resulted in (1) decreased amplitude of cell shortening and of Ca2+ transients, (2) decreased rate of Ca2+ transients decay (3) enhanced diastolic Ca2+ loss. The effect of FK-506 on amplitude of cell shortening was reversed and that on diastolic Ca2+ loss blocked by partial inhibition of NCX due to lowering Na+ concentration in superfusion solution from 144 mM to 100 mM. The amplitude of cell shortening and Ca2+ transients decreased by FK-506 was significantly increased by 10-7 M thapsigargin. In conclusion, the effect of SR Ca2+ leak induced by FK-506 on myocyte contraction is strictly dependent on activity of SERCA and NCX.
The latest developed classes of synthetic insecticides – phthalic acid diamides and antranilic diamides, acting as ryanodine receptor modulators, are discussed. The mode of their activity, the insecticidal properties and characteristics of their representatives is reviewed.
It is widely accepted that Ca2+ is released from the sarcoplasmic reticulum by a specialized type of calcium channel, i.e., ryanodine receptor, by the process of Ca2+-induced Ca2+ release. This process is triggered mainly by dihydropyridine receptors, i.e., L-type (long lasting) calcium channels, directly or indirectly interacting with ryanodine receptor. In addition, multiple endogenous and exogenous compounds were found to modulate the activity of both types of calcium channels, ryanodine and dihydropyridine receptors. These compounds, by changing the Ca2+ transport activity of these channels, are able to influence intracellular Ca2+ homeostasis. As a result not only the overall Ca2+ concentration becomes affected but also spatial distribution of this ion in the cell. In cardiac and skeletal muscles the release of Ca2+ from internal stores is triggered by the same transport proteins, although by their specific isoforms. Concomitantly, heart and skeletal muscle specific regulatory mechanisms are different.
The RYR1 gene encoding the Ca²⁺ channel of sarcoplasmic reticulum of human skeletal muscle has been cloned and its nucleotide sequence has been determined earlier. We have used the polymerase chain reaction single strand conformation polymorphism (PCR-SSCP), and sequencing analysis for human, porcine (Sus scrofa), and zebrine (Equus grevyi) ryanodine receptor (ryrl) gene. The fragment of exon 17 of the ryr1 gene was characterized by a high homology between all the analysed species (substitution of a nucleotide is underlined): porcine ryr1 ¹⁸³⁴GTG GCC GTG CGC TCC AAC CAA GAT CT¹⁸⁵⁹ human RYR1 ¹⁸³¹GTG GCC GTG CGC TCC AAC CAA GAT CT¹⁸⁵⁶ zebrine ryr1 GTG GCC GTG CGC TCC AAC CAA GAC CT.
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Molecular basis of malignant hyperthermia

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Malignant hyperthermia (MH) is a clinical syndrome in which genetically susceptible individuals respond to the administration of potent inhalation anaesthetics and depolarization skeletal muscle relaxants with skeletal rigidity, unstable blood pressure, tachycardia, arrhythmias, hyperventilation, hypoxia, lactic and respiratory acidosis and high fever. In studies of the genetic basis of MH, a mutation was identified in the porcine (C1843T) and human (C1840T) skeletal muscle ryanodine receptor (RYR1) gene. This gene is mapped on human chromosome 19q13.1. The RYR1 gene contains 106 exons, of which two arc alternatively spliced.
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