Abstract
Postsynaptic responses to stereotyped activation of single axons are known to fluctuate, but the origin of synaptic variability in the vertebrate central nervous system is still unclear. To test the hypothesis that fluctuations of inhibitory postsynaptic currents reflect variations in presynaptic Ca2+ concentration, we examined single GABAergic axodendritic contacts in low-density cultures. Collicular neurons from rat embryos were loaded with the Ca2+ indicator Oregon Green 488 BAPTA-1. Presynaptic axon terminals were visualized by staining with the styryl dye RH414. Under the condition of action potential block, RH414-labeled boutons were activated selectively by current pulses applied through a fine-tipped glass pipette. Short (1- to 3-ms) depolarization of isolated boutons resulted in stimulus-locked changes of presynaptic Ca2+ concentration ([Ca2+]pre) and in evoked inhibitory postsynaptic currents (eIPSCs). Varying the strength of the stimulating currents produced a wide amplitude range of both presynaptic fluorescence transients (up to 220% of the resting value) and postsynaptic conductance changes (up to 2-3 nS). It was found that average eIPSCs displayed an approximately third-power dependency on [Ca2+]pre. Transmitter release retained its probabilistic character throughout the range of observed [Ca2+]pre values. In any tested single bouton, maximal eIPSCs occurred in association with the largest [Ca2+]pre transients, but failures were present at any [Ca2+]pre. The increase of maximal eIPSC amplitudes in connection with higher [Ca2+]pre supports the hypothesis that GABAergic boutons have the capacity to regulate synaptic strength by changing the number of simultaneously released vesicles.