<i>Tityus serrulatus</i> venom and its toxins Ts1 and Ts5 increase cytosolic Ca<sup>2+</sup> concentration in isolated vascular smooth muscle cells — Oak Academic Publishing
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<i>Tityus serrulatus</i> venom and its toxins Ts1 and Ts5 increase cytosolic Ca<sup>2+</sup> concentration in isolated vascular smooth muscle cells
Faculty of Ingá-UNINGá, Maringá, Brazil
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Laboratory of Animal Toxins, Department of Physics and Chemistry, Faculty of Pharmaceutical Sciences of Ribeir?o Preto, University of S?o Paulo, Ribeir?o Preto, Brazil
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Laboratory of Pharmacology, Department of Physics and Chemistry, Faculty of Pharmaceutical Sciences of Ribeir?o Preto, University of S?o Paulo, Ribeir?o Preto, Brazil
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Laboratory of Animal Toxins, Department of Physics and Chemistry, Faculty of Pharmaceutical Sciences of Ribeir?o Preto, University of S?o Paulo, Ribeir?o Preto, Brazil
1 Faculty of Ingá-UNINGá, Maringá, Brazil
2 Laboratory of Animal Toxins, Department of Physics and Chemistry, Faculty of Pharmaceutical Sciences of Ribeir?o Preto, University of S?o Paulo, Ribeir?o Preto, Brazil
3 Laboratory of Pharmacology, Department of Physics and Chemistry, Faculty of Pharmaceutical Sciences of Ribeir?o Preto, University of S?o Paulo, Ribeir?o Preto, Brazil
4 Laboratory of Animal Toxins, Department of Physics and Chemistry, Faculty of Pharmaceutical Sciences of Ribeir?o Preto, University of S?o Paulo, Ribeir?o Preto, Brazil
Voltage-gated Na + channel (Nav channel) scorpion toxins are classified as α- and β -neurotoxins. Ts5 ( α -neurotoxin) and Ts1 ( β -neurotoxin) from Tityus serrulatus venom (TsV) interact with Nav channels, increasing Na + influx and activating voltage-dependent Ca 2+ channels. This study aimed to investigate the effect of TsV, Ts1 and Ts5 on the cytosolic Ca 2+ concentration ([Ca 2+ ] C ) in rat aortic smooth muscle cells. Toxins were isolated by ion exchange chromatography (Ts1) followed by RP-HPLC (Ts5). The rat aortic smooth muscle cells were isolated in Hanks buffer pH 7.4 and loaded with 5 μmol/L of Fura-2AM (45 minutes at 37℃), in order to measure [Ca 2+ ] C by fluorescence of Fura-2/AM (ratio 340/380 nm). The fluorescence was measured in one single cell (excitation: 340 and 380 nm; emission: 510 nm). TsV (100 and 500 mg/mL) and its toxins Ts1 and Ts5 (50 and 100 mg/mL each) led to a concentration-dependent increase in [Ca 2+ ] C . Tetrodotoxin (1 mmol/L), a Na v channel blocker, and verapamil (1 mmol/L), a voltage-operated Ca 2+ channel blocker, inhibited the increase in [Ca 2+ ] C induced by TsV (500 mg/mL). In conclusion, TsV and its toxins induce a concentration-dependent increase in [Ca 2+ ] C that probably occurs through interaction with Nav channels, thus inducing depolarization and consequent Ca 2+ influx. This assumption is based on the fact that this effect is inhibited by tetrodotoxin and verapamil, showing a direct action of TsV toxins on aorta smooth muscle cells.
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