Human neutrophils do not express purinergic P2X7 receptors |
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Authors: | Guadalupe Martel-Gallegos María T Rosales-Saavedra Juan P Reyes Griselda Casas-Pruneda Carmen Toro-Castillo Patricia Pérez-Cornejo Jorge Arreola |
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Institution: | 1.School of Medicine,Universidad Autónoma de San Luis Potosí,San Luis Potosí,México;2.Institute of Physics,Universidad Autónoma de San Luis Potosí,San Luis Potosí,México |
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Abstract: | It has been reported that in human neutrophils, external ATP activates plasma membrane purinergic P2X7 receptors (P2X7R) to elicit Ca2+ entry, production of reactive oxygen species (ROS), processing and release of pro-inflammatory cytokines, shedding of adhesion
molecules and uptake of large molecules. However, the expression of P2X7R at the plasma membrane of neutrophils has also been questioned since these putative responses are not always reproduced.
In this work, we used electrophysiological recordings to measure functional responses associated with the activation of membrane
receptors, spectrofluorometric measurements of ROS production and ethidium bromide uptake to asses coupling of P2X7R activation to downstream effectors, immune-labelling of P2X7R using a fluorescein isothiocyanate-conjugated antibody to detect the receptors at the plasma membrane, RT-PCR to determine
mRNA expression of P2X7R and Western blot to determine protein expression in neutrophils and HL-60 cells. None of these assays reported the presence
of P2X7R in the plasma membrane of neutrophils and non-differentiated or differentiated HL-60 cells—a model cell for human neutrophils.
We concluded that P2X7R are not present at plasma membrane of human neutrophils and that the putative physiological responses triggered by external
ATP should be reconsidered. |
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Keywords: | Human Neutrophils Purinergic receptors Protein Whole cell current |
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