Development of Cell-Based Assays for Adenine Receptors and Selected Purine Receptor Subtypes

Abstract

The thesis deals with the development of different cell-based assay techniques in order to characterize the receptors under investigation at the protein level, mapping of the distribution of the receptors of interest in different cells and tissues, studying receptor-ligand interactions and investigating signaling pathways inside the cells subsequent to activation or blocking of these receptors. Such information is important for assessing the receptors' potential as drug targets.<br /> The following assays have been employed: <br /> - Radioligand binding assays including saturation, competition and kinetic experiments in order to characterize the receptors at the protein level and for characterizing novel ligands.<br /> - Functional assays have been established via measurement of cAMP accumulation, measurement of intracellular Ca²⁺ levels and quantification of ERK signalling.<br /> The assays were performed in native cells using intact cell lines or membrane preparations from the receptors under investigation, or at receptors that we have successfully expressed in different cell lines using molecular biology techniques.