Hyperfiltration subjects podocytes to increased tensile stress and fluid flow shear stress (FFSS). We showed a 1.5- to 2.0-fold increase in FFSS in uninephrectomized animals and altered podocyte actin cytoskeleton and increased synthesis of prostaglandin [E.sub.2] (PG[E.sub.2]) following in vitro application of FFSS. We hypothesized that increased FFSS mediates cellular changes through specific receptors of PG[E.sub.2]. Presently, we studied the effect of FFSS on cultured podocytes and decapsulated isolated glomeruli in vitro, and on solitary kidney in uninephrectomized sv129 mice. In cultured podocytes, FFSS resulted in increased gene and protein expression of cyclooxygenase (COX)-2 but not COX-1, prostanoid receptor EP2 but not EP4, and increased synthesis and secretion of PG[E.sub.2], which were effectively blocked by indomethacin. Next, we developed a special flow chamber for applying FFSS to isolated glomeruli to determine its effect on an intact glomerular filtration barrier by measuring change in albumin permeability ([P.sub.alb]) in vitro. FFSS caused an increase in [P.sub.alb], that was blocked by indomethacin (P podocytes; fluid flow shear stress; glomerular hemodynamics; hyperfiltration; glomerular filtration barrier doi: 10.1152/ajprenal.00335.2014.