title: Expression profiling of G-protein-coupled receptors in human urothelium and related cell lines.
abstract: 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.
  
  
  OBJECTIVE
  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.
  
  
  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.
authors.affiliation: Department of Pharmacology and Pharmacotherapy, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands. p.ochodnicky@amc.uva.nl
authors.forename: Peter
authors.initials: P
authors.lastname: Ochodnický
authors.forename: Sian
authors.initials: S
authors.lastname: Humphreys
authors.forename: Rachel
authors.initials: R
authors.lastname: Eccles
authors.forename: Mirjana
authors.initials: M
authors.lastname: Poljakovic
authors.forename: Peter
authors.initials: P
authors.lastname: Wiklund
authors.forename: Martin C
authors.initials: MC
authors.lastname: Michel
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002460:Cell Line; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D043562:Receptors, G-Protein-Coupled / Q000096:biosynthesis*; D019459:Urothelium / Q000166:cytology* / Q000378:metabolism*; D055815:Young Adult
chemical_list: D043562:Receptors, G-Protein-Coupled