AN NMR SPECTROSCOPIC CHARACTERIZATION OF A NEW EPITHELIAL-CELL LINE, TALH-SVE, WITH PROPERTIES OF THE RENAL MEDULLARY THICK ASCENDING LIMB OF HENLES LOOP

NMR spectroscopy has been used to characterize a new renal cell line. TALH-SVE.1, which is derived from the medullary thick ascending limb of Henle's loop. From the P-31-NMR spectrum of a suspension of TALH-SVE cells using the chemical shift of the intracellular inorganic phosphate a value of 7.24 +/- 0.04 for the steady-state intracellular pH (pH(i)) was determined at pH(o) = 7.40. In addition, the P-31-NMR spectrum indicated rather high levels of UDPG, a finding confirmed by H-1-NMR spectra of perchloric acid extracts. The H-1-NMR data also demonstrate the presence of 'organic osmolytes' such as inositol, sorbitol, choline and glycerophosphoryl choline (GPC). C-13-NMR spectra of perchloric acid extracts of TALH-SVE cells incubated with [2-C-13]- and [3-C-13]alanine were used to determine the relative influx in the Krebs cycle via pyruvate carboxylase (PCB) versus the influx via pyruvate dehydrogenase (PDH). The ratio was 0.41, while about 52% of all acetyl-CoA entering the Krebs cycle was unlabeled. C-13-NMR experiments also indicated that TALH-SVE cells lack gluconeogenic activity. The NMR study presented indicates that TALH-SVE cells possess metabolic pathways similar to those of the parental cells.