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Src tyrosine kinases contribute to serotonin-mediated contraction by regulating calcium-dependent pathways in rat skeletal muscle arteries

  • Muscle Physiology
  • Published:
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Abstract

The Src tyrosine kinase family contributes to the signalling mechanism mediating serotonin (5-hydroxytryptamine (5-HT))-induced vasoconstriction. These kinases were reported to influence the calcium sensitivity of the contractile apparatus. Whether Src kinases affect also the intracellular calcium concentration during constriction of intact arteries is unknown. Thus, we tested the hypothesis that constriction of arteries is associated with a Src kinase-dependent alteration of the intracellular calcium concentration. Contractility of gracilis arteries of Wistar rats was studied using isometric and isobaric myography. The intracellular calcium concentration was measured simultaneously with tension by FURA-2 fluorimetry. Inhibition of Src kinases with 10 μM PP2, 30 μM dasatinib and 100 μM AZM 475271 resulted in a strong attenuation of 5-HT-induced contractions. Vessel incubation with 10 μM PP3, an inactive analogue of PP2, had no effect. Removal of the endothelium did not alter vessel contractile responses to 5-HT nor the action of the Src-kinase inhibitor PP2. The PP2-mediated inhibition of 5-HT-induced contraction was associated with a reduced response of [Ca2+]i to 5-HT. In particular, inhibition of Src kinases attenuates 5-HT-induced calcium influx as well as calcium release from intracellular stores. In contrast, the calcium sensitivity of the contractile apparatus and the filling state of the sarcoplasmic reticulum were not influenced by Src kinases during 5-HT-induced contractions. We conclude that Src kinase activation is a powerful mechanism to produce vasoconstriction of small skeletal muscle arteries of rats. This effect is endothelium-independent. The data further suggest that the action of c-Src kinases is associated with a change in the intracellular calcium concentration that involves Ca2+ entry and Ca2+ release pathways.

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Author information

Authors and Affiliations

  1. Centre for Biomedicine and Medical Technology Mannheim (CBTM), Research Division Cardiovascular Physiology, Medical Faculty Mannheim, Heidelberg University, 68167, Mannheim, Germany

    Olga Zavaritskaya, Serdar Altay & Rudolf Schubert

  2. Institute of Vegetative Physiology, University of Cologne, 50931, Cologne, Germany

    Lubomir T. Lubomirov

Authors
  1. Olga Zavaritskaya
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  2. Lubomir T. Lubomirov
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  3. Serdar Altay
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  4. Rudolf Schubert
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Corresponding author

Correspondence to Rudolf Schubert.

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Approval for the use of laboratory animals in this study was granted by a governmental committee on animal welfare.

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The authors declare that they have no conflict of interest.

Electronic supplementary material

Supplemental Fig. 1

a 5-HT-induced diameter change of isobaric vessel preparations in the absence of any additional influence (control) and in the presence of 10 μM PP2 (repeated measures ANOVA: n = 6, p < 0.05). b 5-HT-induced diameter change of isobaric vessel preparations in the absence (control) and presence of 10 μM PP3 (repeated measures ANOVA: n = 6, p = 0.13). c 5-HT-induced change of vessel tension of isometric vessel preparations in the absence (control) and in the presence of 10 μM PP2 (repeated measures ANOVA: n = 6, p < 0.05) d 5-HT-induced change of vessel tension in the absence (control) and in the presence of 10 μM PP3 (repeated measures ANOVA: n = 6, p = 0.39); * - significant difference; n.s. – no significant effect (DOCX 61 kb)

Supplemental Fig. 2

a 5-HT-induced change of vessel tension of isometric vessel preparations in the absence of any additional influence (control) and presence of DMSO, the solvent of PP2 (repeated measures ANOVA: n = 7, p = 0.17). b 5-HT-induced diameter change of isobaric vessel preparations in the absence (control) and presence of DMSO, the solvent of PP2 (repeated measures ANOVA: n = 6, p = 0.56) c 5-HT-induced change of vessel tension of isometric vessel preparations in the absence (control) and presence of 0.3 μM PP2 (repeated measures ANOVA: n = 4, p = 0.56). d 5-HT-induced diameter change of isobaric vessel preparations in the absence (control) and presence of 20 μM PP3 (repeated measures ANOVA: n = 6, p < 0.05); * - significant difference; n.s. – no significant effect (DOCX 60 kb)

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Zavaritskaya, O., Lubomirov, L.T., Altay, S. et al. Src tyrosine kinases contribute to serotonin-mediated contraction by regulating calcium-dependent pathways in rat skeletal muscle arteries. Pflugers Arch - Eur J Physiol 469, 767–777 (2017). https://doi.org/10.1007/s00424-017-1949-3

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  • DOI: https://doi.org/10.1007/s00424-017-1949-3

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