Application of the pyroantimonate method and electron probe microanalysis to the study of glycogen metabolism in liver.

Glycogen distribution in the liver of mouse under different metabolic conditions was studied by the pyroantimonate (PA) method combined with semi-quantitative electron probe microanalysis (EPMA). In the liver of animals subjected to a sugar-rich diet, glycogen granules were abundant and electron transparent. In fasted animals, they were less numerous and stained by PA, which indicates the presence of a complexed cation. This cation was identified as calcium by EPMA. In both cases, adjacent cytoplasmic areas contained "masked" calcium not revealed by PA but detected by EPMA, which is characteristic of a neutral complexed form; but in the case of the fasted animals, the calcium concentration was significantly lower. If the liver of fasted animals was dissected in 0.2% glucose-containing medium, the glycogen areas dramatically released calcium and lost their stainability by PA, whereas mitochondria and adjacent cytoplasm contained many PA precipitates rich in calcium and sodium, suggesting a sudden increase of intracellular [Ca2+]. In mitochondria, the sodium:calcium ratio was relatively constant, which suggests a process involving a coupling between these two elements. Our results could be explained in the light of physiological and biochemical data. We particularly noted that diffusible cations as calcium and sodium did not appear to be displaced over long distances from their likely source. This observation agrees with recent theories on the state of water and ion mobility in the cell.