Functional expression of P2X1, P2X4 and P2X7 purinergic receptors in human monocyte-derived macrophages

• Published in: European journal of pharmacology 2020-12, Vol.888, p.173460-173460, Article 173460
• Main Authors: Vargas-Martínez, Eydie M., Gómez-Coronado, Karen S., Espinosa-Luna, Rosa, Valdez-Morales, Eduardo E., Barrios-García, Tonatiuh, Barajas-Espinosa, Alma, Ochoa-Cortes, Fernando, Montaño, Luis M., Barajas-López, Carlos, Guerrero-Alba, Raquel
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - pharmacology; Animals; ATP; Blood cells; Cells, Cultured; Female; Gene Expression; Human macrophages; Humans; Macrophages - drug effects; Macrophages - metabolism; P2X receptors; Patch-clamp; Purinergic P2X Receptor Agonists - pharmacology; Receptors, Purinergic P2X1 - biosynthesis; Receptors, Purinergic P2X1 - genetics; Receptors, Purinergic P2X4 - biosynthesis; Receptors, Purinergic P2X4 - genetics; Receptors, Purinergic P2X7 - biosynthesis; Receptors, Purinergic P2X7 - genetics; Xenopus laevis
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2020.173460

This study sought to examine the co-expression of the following purinergic receptor subunits: P2X1, P2X1del, P2X4, and P2X7 and characterize the P2X response in human monocyte-derived macrophages (MDMs). Single-cell RT-PCR shows the presence of P2X1, P2X1del, P2X4, and P2X7 mRNA in 40%, 5%, 20%, and 90% of human MDMs, respectively. Of the studied human MDMs, 25% co-expressed P2X1 and P2X7 mRNA; 5% co-expressed P2X4 and P2X7; and 15% co-expressed P2X1, P2X4, and P2X7 mRNA. In whole-cell patch clamp recordings of human MDMs, rapid application of ATP (0.01 mM) evoked fast current activation and two different desensitization kinetics: 1. a rapid desensitizing current antagonized by PPADS (1 μM), reminiscent of the P2X1 receptor's current; 2. a slow desensitizing current, insensitive to PPADS but potentiated by ivermectin (3 μM), similar to the P2X4 receptor's current. Application of 5 mM ATP induced three current modalities: 1. slow current activation with no desensitization, similar to the P2X7 receptor current, present in 69% of human macrophages and antagonized by A-804598 (0.1 μM); 2. fast current activation and fast desensitization, present in 15% of human MDMs; 3. fast activation current followed by biphasic desensitization, observed in 15% of human MDMs. Both rapid and biphasic desensitization kinetics resemble those observed for the recombinant human P2X1 receptor expressed in oocytes. These data demonstrate, for the first time, the co-expression of P2X1, P2X4, and P2X7 transcripts and confirm the presence of functional P2X1, P2X4, and P2X7 receptors in human macrophages.