Quantitative Tc-99m-dimercaptosuccinic acid (DMSA) renal uptake was studied in unilateral reflux-related pyelonephritis in pigs. The changes to absolute % dose uptake and differential uptake occurring with induction and after treatment of pyelonephritis were correlated with the DMSA images and renal pathology. Methods: Quantitative uptake in 53 young pigs was obtained from planar images acquired 6 h after injecting the dose. Baseline studies were made (Q1), and studies were made again after urinary infection was established (Q2), when 8 pigs had normal (no defect) renal images (group A), 23 had photon-deficient (reversible) focal defects (group B) and 22 had photon absent (irreversible) focal defects (group C). Q3 studies were made in 21 animals from groups B and C after 3-wk antimicrobial treatment. Results: At Q2 the affected kidney differential uptake was unchanged for group A and reduced for groups B and C (respective mean changes -1.7%, P < 0.01; and -5.5%, P < 0.01). The absolute % dose uptake was unchanged in pyelonephritic kidneys, but increased in the contralateral nondiseased kidneys in groups B and C (respective mean increases +1.4%, P < 0.05; and +5.4%, P < 0.01), while remaining unchanged for group A, in group C, global renal accumulation was actually increased above the Q1 values. After treatment (Q3) the reduced pyelonephritic kidney differential uptake persisted in groups B and C. In group C, however, the increased absolute % dose uptake by the contralateral kidney was less marked and not significantly different from Q1 values in this small group. Conclusion: Induction of unilateral pyelonephritis produced a small reduction in diseased kidney differential uptake that was greatest in the group with irreversible imaging defects. The method did not discriminate individuals with reversible and irreversible imaging defects. The decrease in pyelonephritic kidney differential uptake resulted from increased DMSA accumulation (absolute % dose uptake) by the nondiseased contralateral kidney, while that in pyelonephritic kidneys remained unchanged. After treatment, the reduced pyelonephritic kidney differential uptake persisted, but the elevated global DMSA accumulation seen for group C (with irreversible imaging defects) was not sustained and was variable.