cGMP-Dependent Protein Kinase Modulates the Sensitivity of Mesenchymal Stromal Cells to Purinergic Agonists
Calcium signaling induced by ATP in mesenchymal stromal cells (MSCs) from the human adipose tissue was studied by using the Ca2+ dye Fluo-4 and Ca2+ imaging. Previously, purinergic agonists have been found to mobilize Ca2+ in the MSC cytoplasm by stimulating IP3 production and triggering Ca2+ release from Ca2+ stores via IP3 receptors. Here we demonstrated that the phosphodiesterase inhibitor IBMX increased a lag period of ATP-induced Ca2+ responses and also shifted the dose–response curve to higher concentrations. When stimulated by ATP at doses close to the threshold concentration, MSCs became unresponsive in the presence of 50 μM IBMX. PKA inhibitor H89 did not restore MSC responsiveness to ATP lost in the presence of IBMX, while the application of KT-5823, a PKG inhibitor, canceled the inhibitory effect of IBMX. Yet, the cell treatment with KT-5823 resulted in a marked decrease in the ATP response delay. Altogether, the data obtained suggest that PKG modulates the MSC sensitivity to ATP, presumably by phosphorylating IP3 receptors in a cGMP-dependent manner.
Keywords:mesenchymal stromal cells purinoreceptors Ca2+ signaling cyclic nucleotides
The work was supported by the Russian Science Foundation (project no. 18-14-00347).
COMPLIANCE WITH ETHICAL STANDARDS
Conflict of interests. The authors declare that they have no conflict of interest.
Statement of compliance with standards of research involving humans as subjects. Informed consent was obtained from all individual participants involved in the study. All procedures were performed in accordance with the order of the RF Ministry of Health of August 11, 2017, no. 517n, and according to the recommendations of the WMA Helsinki Declaration (Declaration of Helsinki: Ethical Principles for Medical Research Involving Human Subjects, including amendments made by the 64th Meeting of WMA in Fortaleza, Brazil, October 2013).
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