An NMR spectroscopic characterization of a new epithelial cell line, TALH-SVE, with properties of the renal medullary thick ascending limb of Henle's 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 31P-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 (pHi) was determined at pHo = 7.40. In addition, the 31P-NMR spectrum indicated rather high levels of UDPG, a finding confirmed by 1H-NMR spectra of perchloric acid extracts. The 1H-NMR data also demonstrate the presence of 'organic osmolytes' such as inositol, sorbitol, choline and glycerophosphoryl choline (GPC). 13C-NMR spectra of perchloric acid extracts of TALH-SVE cells incubated with [2-13C]- and [3-13 C]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. 13C-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.
von Recklinghausen
I R
IR
Max-Planck-Institut für Systemphysiologie, Dortmund F.R.G.
Scott
D M
DM
Jans
A W
AW
D000409:Alanine / Q000494:pharmacology; D000818:Animals; D002460:Cell Line / Q000737:chemistry; D002952:Citric Acid Cycle; D004848:Epithelium / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007679:Kidney Medulla / Q000737:chemistry*; D008138:Loop of Henle / Q000737:chemistry* / Q000201:enzymology / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008954:Models, Biological; D010472:Perchlorates; D010710:Phosphates; D011766:Pyruvate Carboxylase / Q000378:metabolism; D011768:Pyruvate Dehydrogenase Complex / Q000378:metabolism; D011817:Rabbits; D014532:Uridine Diphosphate Glucose / Q000378:metabolism
D010472:Perchlorates; D010710:Phosphates; D011768:Pyruvate Dehydrogenase Complex; D011766:Pyruvate Carboxylase; D000409:Alanine; D014532:Uridine Diphosphate Glucose