Functional hepatocyte cation compartmentation demonstrated with 133Cs NMR.

Abstract

This study utilized the large intrinsic chemical shift range of (133)Cs, a potassium congener, in an NMR study of intracellular cation distribution. It demonstrates two distinct intracellular environments in isolated perfused hepatocytes from cesium-fed rats, evident as compartments with different (133)Cs chemical shifts and containing different proportions of total detected cesium. The chemical shifts of the two intracellular compartments were 2.44 +/- 0.07 and 1.21 +/- 0.18 ppm, relative to the cesium signal from the perfusate. The observation of two distinct intracellular cesium signals suggests slow exchange on an NMR chemical shift time-scale (k exchange > 0.02 s). The area of the high-frequency component represented 62 +/- 10% (N = 12) of the total intracellular cesium signal. Manipulation of the intracellular environment using anoxia with aglycemia or digitonin produced changes in the distribution between the two intracellular compartments, showing their dynamic nature. Changes measured in association with metabolic manipulation suggest cytoplasm and mitochondria as the origin of the high and low-frequency intracellular peaks, respectively.