Kiwamoto H, Ma F H, Higashira H, Park Y C, Kurita T
Department of Urology, Kinki University School of Medicine, Osaka, Japan.
Int J Urol. 2001 Oct;8(10):557-63. doi: 10.1046/j.1442-2042.2001.00370.x.
Muscarinic receptor subtypes of cultured smooth muscle cells from the human bladder body were investigated by the receptor binding assay method. The result was compared with that obtained from the human bladder body tissue to confirm whether the receptor subtypes of the cells are not changed after several passages of cell culture.
Inhibitory effects of various muscarinic antagonists on the binding of [3H]-N-methylscopolamine ([3H]-NMS) to membrane preparations obtained from cultured smooth muscle cells from the fourth subculture of the human bladder body were compared with those prepared from the human bladder body tissue and cells expressing human muscarinic receptor subtypes.
Binding-inhibition constants (pKi) for atropine, pirenzepine, methoctramine, 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP), oxybutynin and propiverine obtained from membrane preparations of cultured smooth muscle cells were 8.91, 6.35, 8.24, 8.53, 7.29 and 5.61, respectively. pKi values of these muscarinic receptor antagonists against the membrane preparation of human bladder body tissue were 9.08, 6.66, 8.05, 8.79, 7.53 and 6.04, respectively. pKi values of cultured smooth muscle cells and tissue from human bladder body were correlated closely with those of insect cells expressing the cloned human M2 receptor subtype.
The binding affinities for various muscarinic receptor antagonists of cultured human smooth muscle cells were maintained through the fourth subculture and it was suggested that the M2 receptor subtype is predominantly expressed in cultured smooth muscle cells of human bladder body as well as in tissue of the human bladder body.
