Calcium ion (Ca(2+)) signaling has been widely implicated in physiological and pathophysiological events in the retina, but little is known about the specific mechanisms utilized by retinal neurons to decrease intracellular Ca(2+). Several cellular mechanisms control intracellular Ca(2+) levels, but their relative significance in mouse retinal ganglion cells is not fully known. We used photometry to measure the dynamics of cytosolic Ca(2+) ([Ca(2+)](i)) clearance after brief, depolarization-induced Ca(2+) entry. Treatment with cyclopiazonic acid, an inhibitor of the sarco-endoplasmic reticulum Ca(2+)-ATPase (SERCA) pumps, significantly slowed the decay of the depolarization-induced Ca(2+) transients. The remaining CPA-insensitive decay was slowed further by inhibition of the plasma membrane Na(+)/Ca(2+) exchanger (NCX) by adding KB-R7943 or SN-6 and by inhibition of the plasma membrane Ca(2+)-ATPase (PMCA) via alkalization of the bath solution. Mitochondrial Ca(2+) uptake contributed little to clearance in retinal ganglion cells. Together, the SERCA, PMCA, and NCX transport mechanisms accounted for up to 85% of calcium clearance under normal conditions. We developed a quantitative model for the dynamic role of removal mechanisms over the physiological change in [Ca(2+)](i). According to our calculations, 15 to 30% of initial Ca(2+) removal is via the SERCA pump, whereas the NCX contributes 30–40% and the PMCA 25–35% of the extrusion.