Mitochondrial granules: are they reliable markers for heavy metal cations?

Mitochondrial granules have been used as markers for heavy metal cations, but since such granules can also be found in tissues in the absence of such cations, an attempt was made to define conditions under which these different granules might be visualised. The tissue used was the smooth muscle of the central ear artery of the rabbit. In all studies, the presence or absence of mitochondrial granules was determined by several observers, using coded specimens so that the previous treatment of the specimens was not at the time known to the observers. Paired tissues were exposed for 30 or 90 minutes at 20, 30 or 39 degrees C to an incubate containing either 10 mmol/l barium or a control barium-free solution. After fixation in osmium tetroxide, there was no difference between the two groups; in both cases granules appeared more frequently the longer the time and the higher the temperature of incubation. In a further series where glutaraldehyde was the fixative, granules were identified in 23 out of 41 tissues incubated with barium, but in only 1 out of 41 control tissues (P less than 0.001). Electron probe microanalysis showed that granules in osmium-fixed tissues contained osmium as the main element, whereas granules in glutaraldehyde-fixed tissues which had been incubated with barium showed barium as the predominant cation. Thus mitochondrial granules can be reliable markers for heavy metal cations, but only under carefully controlled conditions.