A quantitative analysis of purinoceptor expression in the bladders of patients with symptomatic outlet obstruction

Objective To compare the expression of the seven known P2X receptors in hum an bladder from male patients with detrusor instability caused by symptomat ic bladder outlet obstruction with that from control bladders, using a quan titative reverse transcription-polymerase chain reaction (RT-PCR) method. Patients and methods Real-time quantitative RT-PCR provides a system for de tecting and analysing RNA. Bladder biopsies were obtained from nine patient s undergoing prostate surgery and control biopsies were obtained From eight age-matched men undergoing routine bladder endoscopy studies, and who were asymptomatic. Total RNA was extracted from each sample and 10 ng of this u sed for individual PCR reactions. The expression levels of the seven P2X ge nes in the total RNA were then determined. Results In the control bladder, P2X(1) was by far the predominant purinergi c receptor at the RNA level, the remainder consistently present in the orde r P2X(1) >> P2X(4) > P2X(2) >P2X(7) > P2X(5) >> P2X(3) = P2X(6) = 0. Calpon in, a smooth muscle-specific protein, was used as a marker for smooth muscl e content. In bladder from symptomatic patients. the P2X(1)/ calponin ratio was greater than that in controls (P=0.016). There appeared to be no diffe rence in P2X(2), but P2X(4), P2X(5), and P2X(7) were all greater in the sym ptomatic bladder than in the controls. although these differences were not significant. Conclusion P2X(1) is the predominant purinoceptor subtype in the human male bladder, consistent with pharmacological evidence. The amount of P2X(1) re ceptor per smooth muscle cell is greater in the obstructed than in control bladder, suggesting an increase in purinergic function in the unstable blad der arising from bladder outlet obstruction.