| Abstract: | Experiments aimed at the partial reconstitution of the intracellular transport systems regulating the cytosolic free Ca2+ homeostasis are reported. Rat insulinoma subcellular fractions enriched in mitochondria, endoplasmic reticulum (microsomes), and secretory granules were studied. The ambient free Ca2+ concentration maintained by the separate or combined organelles was determined with a Ca2+-selective minielectrode. The data demonstrate that ambient Ca2+] is established by the microsomes, not by the mitochondria or the secretory granules, in the range of resting cytosolic Ca2+ concentrations (0.1-0.2 microM Ca2+). Furthermore, the microsomes are able to deplete largely the mitochondria of their exchangeable calcium. Nonetheless, both mitochondria and microsomes, but not secretory granules, function as a coordinated unit to restore the previous ambient Ca2+] following its perturbation. Thus, mitochondria play a major role in bringing down rapidly ambient Ca2+] to the submicromolar range, whereas the endoplasmic reticulum acts as a relay in the transport mechanisms which lower Ca2+] to the resting level. |
