Cell-matrix interactions modulate transepithelial phosphate transport in Pi-deprived OK cells

In opossum kidney (OK) cells as well as in kidney proximal tubules, Pi depletion increases apical (A) and basolateral (B) Na+- dependent Pi cell influxes. In OK cells' monolayers in contrast to proximal tubules, there is no increase in transepithelial Pi transport. This limitation may be due to altered cell-matrix interactions. A and B cell (32)Pi uptakes and transepithelial 32Pi and [ C-14] mannitol fluxes were measured in OK cells grown on uncoated or on Matrigel-coated filter inserts. Cells were exposed overnight to solution of either low (0.25 mM) or high (2.5 mM) Pi. When grown on Matrigel, immunofluorescence of apical NaPi4 (an isoform of the sodium-phosphate cotransporter) transporters increased and A and B 32Pi uptakes into Pi depleted cells were five and threefold higher than in Pi replete cells (P < 0.001). Pi deprivation resulted in larger increase in A to B (4.6 x, P < 0.001) than in B to A (3.5 x, P < 0.001) Pi flux and net Pi transport from A to B increased 10-fold (P < 0.001). With Pi depletion increases in B to A (3.4 x) and A to B (3.3 x) paracellular [C-14] mannitol fluxes were similar, and its net flux was opposite to that of Pi. In cells grown on uncoated filters, transepithelial and paracellular unidirectional and net Pi fluxes decreased or did not change with Pi depletion, despite twofold increases in apical and basolateral Pi cell influxes. In summary, Matrigel-OK cell interactions, particularly in Pi-depleted cells, led to enhanced expression of apical NaPi4 transporters resulting in higher Pi transport rates across cell boundaries; apical Pi readily entered the transcellular transport pool and paracellular fluxes were smaller fractions of transepithelial Pi fluxes. These Matrigel-induced changes led to an increase in net transepithelial apical to basolateral Pi transport.