Estimates of quantal synaptic parameters in light of more complex vesicle pool models

The subdivision of synaptic vesicles (SVs) into discrete pools is a leading concept of synaptic physiology. To better explain specific properties of transmission and plasticity, it has been suggested initially that the readily releasable pool (RRP) of SVs is subdivided into two parallel pools differing in their release probability. More recently, evidence was provided that sequential pools with a single RRP and a series-connected finite-size replacement pool (RP) inserted between the reserve pool (RSP) and RRP equally well or even better account for most aspects of transmission and plasticity. It was further suggest that a fraction of the presynaptic release sites (N) are initially unoccupied by SVs, with vesicle recruitment occurring rapidly during activity, and furthermore that the number of release sites itself changes with rapid dynamics during activity. Here we propose a framework that identifies specific signs of the presence of the series-connected RP, using a combination of two experimental electrophysiological standard methods, cumulative analysis (CumAna) and multiple probability fluctuation analysis (MPFA). In particular we show that if the y-intercept (y(0)) of CumAna is larger than N reported by MPFA (y(0) > NMPFA) this is a strong indication for a series-connected RP. This is due to the fact that y(0) reports the sum of RRP and RP. Our analysis further suggests that this result is not affected by unoccupied release sites, as such empty sites contribute to both estimates, y(0) and NMPFA. We discuss experimental findings and models in the recent literature in the light of our theoretical considerations.