title: The extraneuronal O-methylation of 3H-(+)isoprenaline by guinea-pig tracheal rings in vitro.
abstract: Tracheal rings isolated from male guinea-pigs and incubated in Krebs' solution at 37 degrees C O-methylated 3H-(+/-)-isoprenaline by a saturable, high affinity mechanism. 1. With 3H-isoprenaline at 1 mumol . 1-1, O-methyl 3H-(+/-)isoprenaline (3H-OMI) appeared in the tissue with a half-time for approach to steady state of approximately 10 min and was measured in the incubation medium after about 5 min, its concentration increasing linearly thereafter. With 3H-isoprenaline concentrations ranging from 1 to 200 mumol . 1-1, the total formation of 3H-OMI (estimated from that contained in the tissue and the medium) was maintained at steady state rates for up to 60 min, after initial lag times of between 1 and 3 min. O-methylation obeyed Michaelis-Menten saturation kinetics; Km = 6.14 +/- 0.13 mumol . 1-1, Vmax = 0.31 +/- 0.01 nmol . g-1 . min-1. 2. The catechol-O-methyl transferase (COMT) inhibitor U-O521 and the extraneuronal uptake inhibitor corticosterone both reduced O-methylation of 3H-isoprenaline (0.1 mumol . 1-1) by tracheal rings. However, U-O521 was fully inhibitory (IC50 = 2.6 mumol . 1-1), but corticosterone inhibited by only 46% at concentrations up to 1 mmol . 1-1. 3. The O-methylating activity of the "smooth muscle-rich" component of the trachea was approximately three-times greater than for complete tracheal rings. However, considerable activity was also associated with "cartilage-rich", "smooth muscle-poor" sections. This activity did not seem to be associated with endothelial cells. 4. Histamine strongly inhibited O-methylation (IC50 = 30 mumol . 1-1), but two other contractile agonists, 5-hydroxytryptamine and bethanechol, were weakly active and inactive, respectively.
mesh_terms: D000818:Animals; D001626:Bethanechol Compounds / Q000494:pharmacology; D006168:Guinea Pigs; D006632:Histamine / Q000494:pharmacology; D066298:In Vitro Techniques; D007545:Isoproterenol / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008745:Methylation; D009130:Muscle, Smooth / Q000378:metabolism*; D011427:Propiophenones / Q000494:pharmacology; D014132:Trachea / Q000378:metabolism*
chemical_list: D001626:Bethanechol Compounds; D011427:Propiophenones; C006104:U 0521; D006632:Histamine; D007545:Isoproterenol