Development of a Radioligand Binding Assay for Detection of Gastrin/CCKB Receptors in the Human Gastrointestinal Tract

[摘要] The initial strategy of the thesis (Chapter 3) examined the presence and characterisation of gastrin/CCKB receptors in the rat pancreatic cell line, AR42J. This cell line was chosen due to its continuous expression of high affinity gastrin/CCKB receptors even after repeated cell culture. Following optimisation of the radioligand binding assay, gastrin/CCKB receptors were characterised using a panel of receptor agonists and antagonists. The AR42J whole cell assay demonstrated that AR42J cells express high affinity gastrin/CCKB receptors with a dissociation constant of 0.3nM and maximal binding capacity of 24fmols/106 cells. These results were similar to those found in the literature by several different groups. Inhibitory dissociation constants (Ki) for the receptor agonists and antagonists used in displacement experiments were also found to correlate closely to literature values thereby confirming the validity of the gastrin/CCKB receptor properties of AR42J cells as measured using the assay developed. The second series of experiments (Chapter 4) examined the preparation of crude membranes from AR42J cells and also the effect of membrane storage. Crude membrane fractions were found to retain the receptor characteristics and properties of receptors on AR42J whole cells. Storage of crude membranes for a limited period at -70

[发布机构] University:University of Glasgow

[关键词] Cellular biology, Oncology