Kinetics of the efflux of 3H-norepinephrine from rabbit irides

The pigment cell dependent accumulation of 3H(-)-norepinephrine (1 mumol/l) was studied in rabbit irides. Albino irides served as controls. Neuronal uptake, deamination and O-methylation of 3H(-)-norepinephrine were reduced by desipramine, pargyline and U-0521, respectively. Both albino and pigmented irides accumulated the catecholamine from the incubation medium. At 37.5 degrees C and 120 min of incubation, tissue/medium ratios for pigmented and albino irides were 10.70 and 4.30 ml/g respectively. At 0 degrees C, the accumulation of 3H(-)-norepinephrine in the albino iris was abolished, while the pigmented iris accumulated significant amounts. The tissue/medium ratio was approximately 3 ml/g. When irides were loaded with 1 mumol/l of 3H(-)-norepinephrine for 60 min the drug distributed into four different compartments. Most of the pigment cell dependent binding was associated with compartments III and the "bound fraction". The amount bound in these compartments was 73% of total accumulation. In pigmented irides obtained from reserpine-pretreated rabbits the total binding was reduced to 64% of that in pigmented control irides. The half times for efflux from pigmented iris were 1.2 min, 10.7 min and 94.8 min, respectively, for compartments I, II, and III. Irides were loaded with 3H(-)-norepinephrine in normal physiological salt solution and rates of efflux were studied in Ca2+-free medium. In either albino or pigmented iris, the late efflux declined more slowly compared to that from the control irides. When pigmented irides were loaded with a low concentration of 3H(-)-norepinephrine, the displacement of the labeled amine was equal by each stereoisomer of norepinephrine. Results are discussed in relationship to the pigment cell-dependent uptake of the catecholamine.