The use of ionic lanthanum as a diffusion tracer and as a marker of calcium binding sites

The distribution of ionic lanthanum (La3+) was studied after perfusion of the isolated rat heart with a buffered salt solution containing lanthanum chloride. We have demonstrated that the localization of lanthanum differed from that shown in previous studies in which hearts were exposed to ionic or colloidal lanthanum during or after fixation. La3+ passed rapidly through the intercellular cleft between endothelial cells. There was relatively little transport by micropinocytosis and no evidence of transendothelial passage through the cytoplasm. Perfusion with a solution containing 1 to 15 mM LaCl3 resulted in localization of La3+ on the entire cytoplasmic membrane system of muscle cells, i.e.: the non-specialized parts of the muscle cell membranes were stained by La3+ as well as the membranes of micropinocytotic vesicles, of the T-system and of the intercalated disc. The cytoplasmic leaflets of gap junctions were especially stained. However, the cell membranes of non-muscle cells were free of lanthanum precipitates. Freeze-fracturing of muscle cells showed concomitant alteration in the distribution of membrane-associated particles. The gap junction membranes also showed changes in particle aggregation. Intracellular binding of La3+ are restricted to muscle cells. Lanthanum was found in the cell nucleus associated with heterochromatin, in the nucleolus, and along the nuclear membrane. The myofibrils were stained mainly over the I-band.