Ca2+ Clearance in Visual Motion-Sensitive Neurons of the Fly Studied In Vivo by Sensory Stimulation and UV Photolysis of Caged Ca2+

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Abstract
In motion-sensitive visual neurons of the fly, excitatory visual stimulation elicits Ca2+ accumulation in dendrites and presynaptic arborizations. Following the cessation of motion stimuli, decay time courses of the cytosolic Ca2+ concentration signals measured with fluorescent dyes were faster in fine arborizations compared with the main branches. When indicators with low Ca2+ affinity were used, the decay of the Ca2+ signals appeared slightly faster than with high affinity dyes, but the dependence of decay kinetics on branch size was preserved. The most parsimonious explanation for faster Ca2+ concentration decline in thin branches compared with thick ones is that the velocity of Ca2+ clearance is limited by transport mechanisms located in the outer membrane and is thus dependent on the neurite's surface-to-volume ratio. This interpretation was corroborated by UV flash photolysis of caged Ca2+ to systematically elicit spatially homogeneous step-like Ca2+ concentration increases of varying amplitude. Clearance of Ca2+ liberated by this method depended on branch size in the same way as Ca2+ accumulated during visual stimulation. Furthermore, the decay time courses of Ca2+ signals were only little affected by the amount of Ca2+ released by photolysis. Thus Ca2+ efflux via the outer membrane is likely to be the main reason for the spatial differences in Ca2+ clearance in visual motion-sensitive neurons of the fly.