Immunofluorescence Staining of P2X7 Receptors in Whole-Mount Myenteric Plexus Preparations

Abstract

P2X7 receptors play an important role in cytokine release and immune cell regulation. Their upregulation has been described in inflammatory and degenerative processes and P2X7 blockade or deletion has been shown to reduce tissue damage and severity of symptoms in animal models of inflammatory bowel disease (IBD). Several studies have found that P2X7 receptors are present on enteric neurons and glia and it was proposed that they mediate neuronal death during IBD. However, the cell type–specific localization of P2X7 receptors has been a matter of debate, since some antibodies have been found to be unspecific. Here we describe the preparation of whole-mount myenteric plexus from the colon of BAC transgenic P2X7-EGFP reporter mice and subsequent immunofluorescence staining of P2X7 receptors together with cell type-specific marker proteins.