Nucleotides-Induced Changes in the Mechanical Properties of Living Endothelial Cells and Astrocytes, Analyzed by Atomic Force Microscopy



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Gil Redondo, Juan Carlos and Iturri, Jagoba and Ortega de la O, Felipe and Pérez Sen, Raquel and Weber, Andreas and Miras Portugal, María Teresa and Toca Herrera, José Luis and García Delicado, Esmerilda (2021) Nucleotides-Induced Changes in the Mechanical Properties of Living Endothelial Cells and Astrocytes, Analyzed by Atomic Force Microscopy. International Journal of Molecular Sciences, 22 (2). p. 624. ISSN 1422-0067

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Endothelial cells and astrocytes preferentially express metabotropic P2Y nucleotide receptors, which are involved in the maintenance of vascular and neural function. Among these, P2Y1 and P2Y2 receptors appear as main actors, since their stimulation induces intracellular calcium mobilization and activates signaling cascades linked to cytoskeletal reorganization. In the present work, we have analyzed, by means of atomic force microscopy (AFM) in force spectroscopy mode, the mechanical response of human umbilical vein endothelial cells (HUVEC) and astrocytes upon 2MeSADP and UTP stimulation. This approach allows for simultaneous measurement of variations in factors such as Young’s modulus, maximum adhesion force and rupture event formation, which reflect the potential changes in both the stiffness and adhesiveness of the plasma membrane. The largest effect was observed in both endothelial cells and astrocytes after P2Y2 receptor stimulation with UTP. Such exposure to UTP doubled the Young’s modulus and reduced both the adhesion force and the number of rupture events. In astrocytes, 2MeSADP stimulation also had a remarkable effect on AFM parameters. Additional studies performed with the selective P2Y1 and P2Y13 receptor antagonists revealed that the 2MeSADP-induced mechanical changes were mediated by the P2Y13 receptor, although they were negatively modulated by P2Y1 receptor stimulation. Hence, our results demonstrate that AFM can be a very useful tool to evaluate functional native nucleotide receptors in living cells.

Item Type:Article
Uncontrolled Keywords:atomic force microscopy; astrocytes; endothelial cells; nucleotide receptor; P2Y nucleotide receptor
Subjects:Medical sciences > Medicine > Biochemistry
Medical sciences > Medicine > Neurosciences
Medical sciences > Biology > Cytology
Medical sciences > Biology > Neurosciences
ID Code:71671
Deposited On:07 Apr 2022 13:35
Last Modified:11 Apr 2022 10:19

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