Calcium-ion transport by intact synaptosomes. Intrasynaptosomal compartmentation and the role of the mitochondrial...

The association of Ca2+ with isolated nerve endings (synaptosomes) is investigated and resolved into two components, that bound to the outer surface of the plasma membrane and that transported across the plasma membrane. When synaptosomes are added directly to a Ca2+-containing medium, there is an initial rapid uptake of Ca2+ across the plasma membrane, followed by a slow uptake that proceeds for 20 min. The rapid phase is not observed if the synaptosomes are initially pre-incubated in a Ca2+-free medium. Rapid disruption of synaptosomes reveals that less than 3 nmol of transported Ca2+ per mg of synaptosomal protein can be ascribed to non-mitochondrial components, whereas the remainder, up to 79% of the total, is further transported into the mitochondrial matrix. Abolition of oxidative phosphorylation while the mitochondrial membrane potential is retained leads to a time-dependent increase in transported Ca2+, whereas abolition of the mitochondrial membrane potential decreases both plasma-membrane transport and accumulation of Ca2+ in the mitochondrial matrix. It is concluded that intrasynaptosomal mitochondria are major regulators of synaptosomal Ca2+.