The fractional clearances (theta) for FITC-Ficoll and albumin were estimated in isolated perfused rat kidneys in which the tubular activity was inhibited by low temperature (8 degrees C) and/or 10 mM NH(4)Cl. The Ficoll data were analyzed according to a two-pore model giving small and large pore radii of 46 A and 80-87 A, respectively. The estimated negative charge density was 35-45 meq/l at 8 degrees C. Perfusion with erythrocyte-free solutions of kidneys at 37 degrees C reduced glomerular size and charge permselectivity. Thus the large pore fraction of the glomerular filtrate (f(L)) was 1.64% at 37 degrees C compared with 0.94% at 8 degrees C. The theta for albumin was four times higher at 37 degrees C than at 8 degrees C (0.86% vs. 0.19%, respectively). NH(4)Cl caused further irreversible damage to the glomerular barrier. We conclude that there are no deleterious effects on the glomerular barrier of a reduction in temperature from 37 degrees C to 8 degrees C. Therefore our data seem to disprove the hypothesis of low glomerular permselectivity and transtubular uptake of intact albumin and support the classic concept of a highly selective glomerular barrier.