Human neutrophils do not express purinergic P2X7 receptors

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• 作者：Guadalupe Martel-Gallegos ; Mar¨ªa T. Rosales-Saavedra ; Juan P. Reyes ; Griselda Casas-Pruneda ; Carmen Toro-Castillo ; Patricia P¨¦rez-Cornejo and Jorge Arreola
• 关键词：Human ; Neutrophils ; Purinergic receptors ; Protein ; Whole cell current
• 刊名：Purinergic Signalling
• 出版年：2010
• 出版时间：September 2010
• 年：2010
• 卷：6
• 期：3
• 页码：297-306
• 全文大小：389.4 KB

文摘

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.