Rat kidneys subjected to urinary obstruction or dehydration in vivo both develop marked increases in the area under a peak resonating in the phosphodiester region on their P-31 nuclear magnetic resonance spectra. The chemical species responsible for these changes were assessed using physiologic manipulations which altered the concentration of phosphate in the urine or increased the urine pH. Obstructed kidneys of rats fed a normal diet had a 140 +/- 50% increase in a peak resonating at 3.31 +/- 0.05 ppm after three hours of obstruction (P less than 0.01). Low phosphate diet which decreased urine phosphate concentration by 98% virtually eliminated this increase in peak area, where saline diuresis which decreased urine phosphate concentration by 50% markedly attenuated it. Acute phosphate loading which doubled urine phosphate concentration markedly accentuated the increase in peak area. Alkalinizing the urine with acetazolamide (changing urine pH from 6.2 +/- 0.2 to 8.0 +/- 0.1) shifted the resonance frequency of this increasing peak from 3.31 +/- 0.06 to 5.45 +/- 0.11 ppm (P less than 0.01). Rats fed a normal diet developed increases (57 +/- 15%, P less than 0.05) in a peak resonating at 2.84 +/- 0.03 ppm following 48 hours of dehydration. Rats fed a low phosphate diet had a comparable increase in the relative area of this peak (46 +/- 16%, P less than 0.05). Alkalinization of the urine did not affect the position or intensity of this peak under conditions of dehydration.(ABSTRACT TRUNCATED AT 250 WORDS)