PURINERGIC RECEPTOR RESEARCH

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Epithelial cells along the collecting duct regulate free water reabsorption and acid secretion through various hormones and neurotransmitters, including atrial natriuretic peptide, bradykinin, catecholamines, miner-alocorticoids, prostanoids, and vasopressin. Under certain circumstances, P2-purinoceptor agonists (ATP, UTP, ADP, and UDP) can also regulate ion transport in these cells by activating various signaling cascades. Two types of P2-purinoceptors have been described to date: P2X-purinoceptors that correspond to ligand-gated cation channels and P2Y-purinoceptors that are coupled to heterotrimeric G proteins. Both the P2X- and P2Y-purinoceptors are known to occur in several isoforms, most of which exhibit wide tissue distribution. These proteins are also diffusely distributed along various nephron segments. Previous work suggests striking differences in expression of G protein coupled receptors on polarized MDCK cells but information regarding endogenously expressed purinergic P2Y receptor subtypes is not available. We are using the Bio-rad Radiance confocal microscope and Imaris software in order to examine the localization of various purinergic receptors in MDCK D-1 cell in order to detect green fluorescent protein (GFP) labeled P2Y purinergic receptors. Specifically, we are attempting to image sagital and transverse images of confluent cell layers.