The sidedness (apical vs basolateral) of the inhibitory of phosphate (Pi) transport by parathyroid hormone (PTH) was investigated in opossum kidney (OK)-cell monolayers grown on permeant support. PTH was found to regulate the activity of only the apical Na Pi cotransporter, having no effect on the basolateral transport systems. Transport inhibition was approximately 100-fold more sensitive to apical PTH application (Kd: 5 x 10(-12) M) than to basolateral application (Kd: 5 x 10(-10) M). The time-course of the inhibitory response was identical from the two cell surfaces, with half-maximum inhibition occurring at about 20 min and almost full inhibition by 90 min. Experiments on diffusion and degradation demonstrated that the difference in Kd at the two cell surfaces was not due to differential metabolism or diffusion. Tests of cooperativity between the apical and basolateral regulatory events at intermediate concentrations suggested that the presence of PTH on one side of the monolayer reduced the scope of response from the other side. At maximum doses of PTH (10(-7)-10(-8) M) the transport inhibition from either side was equal and not additive. We conclude that in OK-cell monolayers grown on permeant support only apical Na/Pi co-transport is sensitive to PTH inhibition and that PTH receptor properties may be different on the apical and basolateral surfaces.
Functional asymmetry in phosphate transport and its regulation in opossum kidney cells: parathyroid hormone inhibition.
RESHKIN, Stephan Joel;
1990-01-01
Abstract
The sidedness (apical vs basolateral) of the inhibitory of phosphate (Pi) transport by parathyroid hormone (PTH) was investigated in opossum kidney (OK)-cell monolayers grown on permeant support. PTH was found to regulate the activity of only the apical Na Pi cotransporter, having no effect on the basolateral transport systems. Transport inhibition was approximately 100-fold more sensitive to apical PTH application (Kd: 5 x 10(-12) M) than to basolateral application (Kd: 5 x 10(-10) M). The time-course of the inhibitory response was identical from the two cell surfaces, with half-maximum inhibition occurring at about 20 min and almost full inhibition by 90 min. Experiments on diffusion and degradation demonstrated that the difference in Kd at the two cell surfaces was not due to differential metabolism or diffusion. Tests of cooperativity between the apical and basolateral regulatory events at intermediate concentrations suggested that the presence of PTH on one side of the monolayer reduced the scope of response from the other side. At maximum doses of PTH (10(-7)-10(-8) M) the transport inhibition from either side was equal and not additive. We conclude that in OK-cell monolayers grown on permeant support only apical Na/Pi co-transport is sensitive to PTH inhibition and that PTH receptor properties may be different on the apical and basolateral surfaces.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
