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Expression profiling of G‐protein‐coupled receptors in human urothelium and related cell lines
Author(s) -
Ochodnický Peter,
Humphreys Sian,
Eccles Rachel,
Poljakovic Mirjana,
Wiklund Peter,
Michel Martin C.
Publication year - 2012
Publication title -
bju international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.773
H-Index - 148
eISSN - 1464-410X
pISSN - 1464-4096
DOI - 10.1111/j.1464-410x.2012.011145.x
Subject(s) - urothelium , urothelial cell , biology , g protein coupled receptor , receptor , microbiology and biotechnology , cancer research , signal transduction , endocrinology , genetics , urinary system
What's known on the subject? and What does the study add? Urothelium emerged as a crucial integrator of sensory inputs and outputs in the bladder wall, and urothelial G‐protein‐coupled receptors (GPCRs) may represent plausible targets for treatment of various bladder pathologies. Urothelial cell lines provide a useful tool to study urothelial receptor function, but their validity as models for native human urothelium remains unclear. We characterize the mRNA expression of genes coding for GPCRs in human freshly isolated urothelium and compare the expression pattern with those in human urothelial cell lines. OBJECTIVES• To characterize the mRNA expression pattern of genes coding for G‐protein‐coupled receptors (GPCRs) in human freshly isolated urothelium. • To compare GPCR expression in human urothelium‐derived cell lines to explore the suitability of these cell lines as model systems to study urothelial function.MATERIALS AND METHODS• Native human urothelium (commercially sourced) and human urothelium‐derived non‐cancer (UROtsa and TERT‐NHUC) and cancer (J82) cell lines were used. • For mRNA expression profiling we used custom‐designed real‐time polymerase chain reaction array for 40 receptors and several related genes.RESULTS• Native urothelium expressed a wide variety of GPCRs, including α 1A , α 1D and all subtypes of α 2 and β adrenoceptors. In addition, M 2 and M 3 cholinergic muscarinic receptors, angiotensin II AT 1 receptor, serotonin 5‐HT 2A receptor and all subtypes of bradykinin, endothelin, cannabinoid, tachykinin and sphingosine‐1‐phosphate receptors were detected. Nerve growth factor and both its low‐ and high‐affinity receptors were also expressed in urothelium. • In all cell lines expression of most GPCRs was markedly downregulated, with few exceptions. • In UROtsa cells, but much less in other cell lines, the expression of β 2 adrenoceptors, M 3 muscarinic receptors, B 1 and B 2 bradykinin receptors, ET B endothelin receptors and several subtypes of sphingosine‐1‐phosphate receptors was largely retained.CONCLUSIONS• Human urothelium expresses a wide range of receptors which enables sensing and integration of various extracellular signals. • Human urothelium‐derived cell lines, especially UROtsa cells, show comparable mRNA expression to native tissue for several physiologically relevant GPCRs, but lose expression of many other receptors. • The use of cell lines as model systems of human urothelium requires careful validation of suitability for the genes of interest.