Maturation of Schaffer collateral synapses generates a phenotype of unreliable basal evoked release and very reliable facilitated release.

Short-term synaptic plasticity undergoes important age-dependent changes that have crucial implications during the development of the nervous system. Paired-pulse facilitation is a form of short-term synaptic plasticity by which the response to the second of two temporally-paired stimuli is larger and more reliable than the response to the first stimulus. In this study, a paired-pulse minimal stimulation technique was used to measure the probability and quantal amplitude of synaptic release at hippocampal synapses from 12-16-day-old (young) and 7-9-week-old (adult) rats. In order to assess the contribution of temperature-dependent processes, we carried out experiments at both room temperature and at near physiological temperature. We report here that neither temperature nor maturation affected the low basal evoked release probability and quantal amplitude of release. However, the warmer temperature revealed a unique developmental increase in facilitated evoked release probability and quantal amplitude of release. As a result, although both basal evoked release and facilitated release are rather unreliable in synapses from young animals, the maturation process at near physiological temperature generates a phenotype with unreliable basal evoked release and highly reliable facilitated release.

Facilitated glutamate release at Schaffer collateral to CA1 synapses has access to an exclusive population of NMDA receptors.

In order to explore short-term facilitation of the Schaffer collateral to CA1 synapse in mouse hippocampal brain slices, we measured the time course of the decay of the peak amplitude of successive EPSCs during progressive MK-801-dependent block (PMDB) of NMDAR responses to paired (R1 and R2) stimuli. We made the unexpected observation that the R2 response exhibited a slower PMDB decay constant than that of the R1 response. This indicated that the facilitated R2 response engages release sites with NMDARs that are protected from opening and consequent MK-801 block during the basal R1 response. We then utilized conditions that affect synaptic glutamate distribution to dissect the components of the distinct PMDB decay constants of the first and second of paired pulses. While extra-synaptic NMDARs and glutamate transporters appear to play only minor roles in the differences of the PMDB decay constant, we showed important roles for the R1 response itself and for glutamate diffusion in determining the PMDB decay constant of R2. We used a simple computational model with realistic parameters that allowed us to predict the time course of R2 decay based on the R1 decay time course.

Neurosteroid-induced enhancement of short-term facilitation involves a component downstream from presynaptic calcium in hippocampal slices.

We used Magnesium Green AM to measure Ca(2+) transients in Schaffer collateral presynaptic terminals simultaneously with postsynaptic field potentials (fEPSPs) to investigate the mechanism of neurosteroid enhancement of short-term synaptic facilitation. Measurement of [Ca(2+)](i), isolated to presynaptic events, using the fluorescence ratio (DeltaF/F(0)) demonstrated that at a constant stimulus intensity there was no change in the excitability of presynaptic fibres between paired stimuli or between ACSF and 1 mum pregnenolone sulphate (PREGS). Paired-pulse facilitation (PPF) was correlated with residual Ca(2+) ([Ca(2+)](res)), and there was an additional increase in the integralDeltaF/F(0) for the [Ca(2+)](res)-subtracted response to the second of paired stimuli, resulting primarily from a slowing of the decay time constant. In addition to the role of presynaptic [Ca(2+)](res) in PPF, we observed a decrease in EC(50) and a greater maximum for Hill function fits to fEPSP versus DeltaF/F(0) during the second of paired responses. The enhancement of fEPSP PPF by PREGS did not result from an increase of DeltaF/F(0). The data presented here support a PREGS-induced increase in presynaptic glutamate release from the second, but not the first, of a pair of stimuli for a given presynaptic [Ca(2+)] because: (a) there is actually a decrease in the integralDeltaF/F(0) of the [Ca(2+)](res)-subtracted second response over that seen in ACSF; (b) PREGS causes no change in presynaptic Ca(2+) buffering; and (c) there is a decrease in EC(50) and an increase of y(max) in the Hill function fits to DeltaF/F(0) versus fEPSP data. We hypothesize that PREGS enhances short-term facilitation by acting on the Ca(2+)-dependent vesicle release machinery and that this mechanism plays a role in the cognitive effects of this sulphated neurosteroid.