• cerebellum
• molecular layer interneurons
• mouse
• neuroscience
• parallel fibers
• synaptic terminals
• synaptic vesicles

• Animals
• Mice
• Action Potentials
• Binding Sites
• Cerebellum
• Models, Statistical
• Synapses
• Mammals

• 294509 European Research Council
• Fondation pour la Recherche Médicale

• cAMP-mediated potentiation
• hippocampal mossy fiber synapses
• presynaptic plasticity
• readily-releasable pool
• synaptic vesicle release

• Japan Society for the Promotion of Science
• Takeda Science Foundation

Central nervous system synapses are diverse in strength and plasticity. Short-term plasticity has traditionally been evaluated with models postulating a single pool of functionally homogeneous fusion-competent synaptic vesicles. Many observations are not easily explainable by such simple models. We established and experimentally validated a scheme of synaptic vesicle priming consisting of two sequential and reversible steps of release–machinery assembly. This sequential two-step priming scheme faithfully reproduced plasticity at a glutamatergic model synapse. The proposed priming and fusion scheme was consistent with the measured mean responses and with the experimentally observed heterogeneity between synapses. Vesicle fusion probability was found to be relatively uniform among synapses, while the priming equilibrium at rest of mature versus immature vesicle priming states differed greatly.

Glutamatergic synapses display variable strength and diverse short-term plasticity (STP), even for a given type of connection. Using nonnegative tensor factorization and conventional state modeling, we demonstrate that a kinetic scheme consisting of two sequential and reversible steps of release–machinery assembly and a final step of synaptic vesicle (SV) fusion reproduces STP and its diversity among synapses. Analyzing transmission at the calyx of Held synapses reveals that differences in synaptic strength and STP are not primarily caused by variable fusion probability (p_fusion) but are determined by the fraction of docked synaptic vesicles equipped with a mature release machinery. Our simulations show that traditional quantal analysis methods do not necessarily report p_fusion of SVs with a mature release machinery but reflect both p_fusion and the distribution between mature and immature priming states at rest. Thus, the approach holds promise for a better mechanistic dissection of the roles of presynaptic proteins in the sequence of SV docking, two-step priming, and fusion. It suggests a mechanism for activity-induced redistribution of synaptic efficacy.

• calyx of Held
• numerical simulation
• short-term plasticity
• synaptic transmission
• synaptic vesicle priming

Short-term changes in the strength of synaptic connections underlie many brain functions. The strength of a synapse in response to subsequent stimulation is largely determined by the remaining number of synaptic vesicles available for release. We developed a methodological approach to measure the dynamics of various vesicle pools following synaptic activity. We find that the readily releasable pool, which comprises vesicles that are docked or tethered to release sites, is fed by a small-sized pool containing approximately one to four vesicles per release site at rest. This upstream pool is significantly depleted even after a short stimulation train. Therefore, regulation of the size of the upstream pool emerges as a key factor in determining synaptic strength during and after sustained stimulation.

During prolonged trains of presynaptic action potentials (APs), synaptic release reaches a stable level that reflects the speed of replenishment of the readily releasable pool (RRP). Determining the size and filling dynamics of vesicular pools upstream of the RRP has been hampered by a lack of precision of synaptic output measurements during trains. Using the recent technique of tracking vesicular release in single active zone synapses, we now developed a method that allows the sizes of the RRP and upstream pools to be followed in time. We find that the RRP is fed by a small-sized pool containing approximately one to four vesicles per docking site at rest. This upstream pool is significantly depleted by short AP trains, and reaches a steady, depleted state for trains of >10 APs. We conclude that a small, highly dynamic vesicular pool upstream of the RRP potently controls synaptic strength during sustained stimulation.

• C2 domains
• Calcium
• Munc13
• Phospholipids
• Presynapse
• Short-termsynaptic plasticity
• Synapse
• Synaptic transmission
• Synaptic vesicle replenishment
• calyx of Held

• Action Potentials
• Animals
• Calcium/metabolism
• Mice
• Neuronal Plasticity/physiology
• Phospholipids/metabolism
• Synaptic Transmission/physiology
• Synaptic Vesicles/metabolism

• bioenergetics
• glycolysis
• mouse
• oxidative phosphorylation
• synaptic vesicle cycle

• Animals
• Cochlear Nucleus/metabolism
• Female
• Glycolysis/physiology
• Male
• Mice
• Mice, Inbred C57BL
• Mitochondria/metabolism
• Respiration
• Synapses/metabolism
• Synaptic Transmission/physiology

• P20 GM103554 NIGMS NIH HHS
• R01 NS117686 NINDS NIH HHS
• R01 DC019268 NIDCD NIH HHS
• HHS | NIH | National Institute of General Medical Sciences (NIGMS)
• HHS | NIH | National Institute of Neurological Disorders and Stroke (NINDS)
• HHS | NIH | National Institute on Deafness and Other Communication Disorders (NIDCD)
• NSF | BIO | Division of Molecular and Cellular Biosciences (MCB)

• presynapse
• presynaptic development
• synapse

Action potentials trigger two modes of neurotransmitter release, with a fast synchronous component and a temporally delayed asynchronous release. Asynchronous release contributes to information transfer at synapses, including at the hippocampal mossy fiber (MF) to CA3 pyramidal cell synapse where it controls the timing of postsynaptic CA3 pyramidal neuron firing. Here, we identified and characterized the main determinants of asynchronous release at the MF–CA3 synapse. We found that asynchronous release at MF–CA3 synapses can last on the order of seconds following repetitive MF stimulation. Elevating the stimulation frequency or the external Ca²⁺ concentration increased the rate of asynchronous release, thus, arguing that presynaptic Ca²⁺ dynamics is the major determinant of asynchronous release rate. Direct MF bouton Ca²⁺ imaging revealed slow Ca²⁺ decay kinetics of action potential (AP) burst‐evoked Ca²⁺ transients. Finally, we observed that asynchronous release was preferentially mediated by Ca²⁺ influx through P/Q‐type voltage‐gated Ca²⁺ channels, while the contribution of N‐type VGCCs was limited. Overall, our results uncover the determinants of long‐lasting asynchronous release from MF terminals and suggest that asynchronous release could influence CA3 pyramidal cell firing up to seconds following termination of granule cell bursting.

• asynchronous release
• calcium
• hippocampus
• mossy fibers
• presynaptic

• Animals
• Calcium/metabolism
• Cerebellum/physiology
• Interneurons/physiology
• Presynaptic Terminals/physiology
• Rats
• Rats, Sprague-Dawley
• Synapses/physiology
• Synaptic Transmission/physiology
• Synaptic Vesicles/physiology

The efficacy of neocortical synapses to transmit during bursts of action potentials (APs) increases during development but the underlying mechanisms are largely unclear. We investigated synaptic efficacy at synapses between layer 5 pyramidal neurons (L5PNs) during development, using paired recordings, presynaptic two-photon Ca²⁺ imaging, and numerical simulations. Our data confirm a developmental increase in paired-pulse ratios (PPRs). Independent of age, Ca²⁺ imaging revealed no AP invasion failures and linear summation of presynaptic Ca²⁺ transients, making differences in Ca²⁺ signaling an unlikely reason for developmental changes in PPR. Cumulative excitatory postsynaptic current (EPSC) amplitudes indicate that neither the size of the readily-releasable pool (RRP) nor replenishment rates were different between age groups, while the time-courses of depression differed significantly. At young synapses, EPSCs depressed rapidly to near steady-state during the first four APs, and synaptic failures (F_syn) increased from 0 to 30%. At mature synapses this drop was significantly slower and strongly biphasic, such that near steady-state depression was reached not before 18 APs with F_syn remaining between 0 and 5%. While young synapses reliably transmitted during pairs of APs, albeit with strong depression, mature synapses maintained near 100% transfer efficacy with significantly less depression during high-frequency bursts of APs. Our analysis indicates that at mature synapses a replenishment pool (RepP) is responsible for their high efficacy during bursting activity, while this RepP is functionally immature at young synapses. Hence, our data provide evidence that the functional maturation of a RepP underlies increasing synaptic efficacy during the development of an excitatory cortical synapse.

• action potential trains
• failures
• layer 5 pyramidal neurons
• paired-pulse ratios
• presynaptic calcium signals
• replenishment
• synaptic efficacy
• vesicle pools

• Presynaptic
• Synapse
• exocytosis
• synaptic transmission

• Basal ganglia
• Cortex
• Glutamate
• Synapse
• Thalamus

• neurotransmitter release
• quantal analysis
• release site
• synapse
• vesicle recruitment

• GPCR
• action potential
• modulation
• neurotransmission
• presynaptic

• R01 DA035913 NIDA NIH HHS
• R01 MH112729 NIMH NIH HHS
• R21 MH112117 NIMH NIH HHS

• RRP
• STP
• mouse
• neuroscience
• short-term plasticity
• vesicle

The release of neurotransmitters from synapses obeys complex and stochastic dynamics. Depending on the recent history of synaptic activation, many synapses depress the probability of releasing more neurotransmitter, which is known as synaptic depression. Our understanding of how synaptic depression affects the information efficacy, however, is limited. Here we propose a mathematically tractable model of both synchronous spike-evoked release and asynchronous release that permits us to quantify the information conveyed by a synapse. The model transits between discrete states of a communication channel, with the present state depending on many past time steps, emulating the gradual depression and exponential recovery of the synapse. Asynchronous and spontaneous releases play a critical role in shaping the information efficacy of the synapse. We prove that depression can enhance both the information rate and the information rate per unit energy expended, provided that synchronous spike-evoked release depresses less (or recovers faster) than asynchronous release. Furthermore, we explore the theoretical implications of short-term synaptic depression adapting on longer time scales, as part of the phenomenon of metaplasticity. In particular, we show that a synapse can adjust its energy expenditure by changing the dynamics of short-term synaptic depression without affecting the net information conveyed by each successful release. Moreover, the optimal input spike rate is independent of the amplitude or time constant of synaptic depression. We analyze the information efficacy of three types of synapses for which the short-term dynamics of both synchronous and asynchronous release have been experimentally measured. In hippocampal autaptic synapses, the persistence of asynchronous release during depression cannot compensate for the reduction of synchronous release, so that the rate of information transmission declines with synaptic depression. In the calyx of Held, the information rate per release remains constant despite large variations in the measured asynchronous release rate. Lastly, we show that dopamine, by controlling asynchronous release in corticostriatal synapses, increases the synaptic information efficacy in nucleus accumbens.

Fatigue is an intrinsic property of living systems and synapses are no exception. Synaptic depression reduces the ability of synapses to release vesicles in response to an incoming action potential. Whether synaptic depression simply reflects the exhaustion of neuronal resources or whether it serves some additional function is still an open question. We ask how synaptic depression modulates the information transfer between neurons by keeping the synapse in an appropriate operating range. Using a tractable mathematical model for synaptic depression of both synchronous spike-evoked and asynchronous release of neurotransmitter, we derive a closed-form expression for the mutual information rate. Depression, it turns out, can both enhance or impair information transfer, depending on the relative level of depression for synchronous spike-evoked and asynchronous releases. We also study the compromise a synapse makes between its energy consumption and the rate of information transmission. Interestingly, we show that synaptic depression can regulate energy use without affecting the information (measured in bits) per synaptic release. By applying our mathematical framework to experimentally measured synapses, we show that some synapses can compensate for intrinsic variability in asynchronous release rates; moreover, we show how neuromodulators such as dopamine act to improve the information transmission rate.

• Synapse
• calcium buffering
• short-term plasticity
• vesicle recruitment

Established models of vesicular docking/release sites can account for synaptic depression. By incorporating a separate predocked state and by assuming that docking site occupancy is <1 at rest, Pulido and Marty extend previous models and explain facilitating and nonmonotonic synaptic responses.

The strength of synaptic transmission varies during trains of presynaptic action potentials, notably because of the depletion of synaptic vesicles available for release. It has remained unclear why some synapses display depression over time, whereas others facilitate or show a facilitation and depression sequence. Here we compare the predictions of various synaptic models assuming that several docking/release sites are acting in parallel. These models show variation of docking site occupancy during trains of action potentials due to vesicular release and site replenishment, which give rise to changes in synaptic strength. To conform with recent studies, we assume an initial docking site occupancy of <1, thus permitting site occupancy to increase during action potential trains and facilitation to occur. We consider both a standard one-step model and a more elaborate model that assumes a predocked state (two-step model). Whereas the one-step model predicts monotonic changes of synaptic strength during a train, the two-step model allows nonmonotonic changes, including the often-observed facilitation/depression sequence. Both models predict a partitioning of parameter space between initially depressing and facilitating synapses. Using data obtained from interneuron synapses in the cerebellum, we demonstrate an unusual form of depression/facilitation sequence for very high release probability after prolonged depolarization-induced transmitter release. These results indicate a depletion of predocked vesicles in the two-step model. By permitting docking site occupancy to be <1 at rest, and by incorporating a separate predocked state, we reveal that docking site models can be expanded to mimic the large variety of time-dependent changes of synaptic strength that have been observed during action potential trains. Furthermore, the two-step model provides an effective framework to identify the specific mechanisms responsible for short-term changes in synaptic strength.

• Corticofugal axon
• Pyramidal neuron
• Subthalamic nucleus

• active zone
• replenishment
• synaptic vesicles

• exocytosis
• presynaptic
• synapse
• synaptic transmission
• transmitter release

• Animals
• Auditory Cortex/cytology
• Humans
• Kinetics
• Neurotransmitter Agents/metabolism
• Synapses/metabolism
• Synaptic Vesicles/metabolism

• Active zone
• Endocytosis
• Exocytosis
• Release site maintenance
• Synapse

The segregation of the readily releasable pool of synaptic vesicles (RRP) in sub-pools that are differentially poised for exocytosis shapes short-term plasticity. However, the frequency-dependent mobilization of these sub-pools is poorly understood. Using slice recordings and modeling of synaptic activity at cerebellar granule cell to Purkinje cell synapses of mice, we describe two sub-pools in the RRP that can be differentially recruited upon ultrafast changes in the stimulation frequency. We show that at low-frequency stimulations, a first sub-pool is gradually silenced, leading to full blockage of synaptic transmission. Conversely, a second pool of synaptic vesicles that cannot be released by a single stimulus is recruited within milliseconds by high-frequency stimulation and support an ultrafast recovery of neurotransmitter release after low-frequency depression. This frequency-dependent mobilization or silencing of sub-pools in the RRP in terminals of granule cells may play a role in the filtering of sensorimotor information in the cerebellum.

• Purkinje cell
• cerebellum
• mouse
• neuroscience
• neurotransmitter release
• parallel fibers
• short-term plasticity
• synaptic vesicles

• biophysics
• calcium channels
• exocytosis
• mouse
• neuroscience
• presynapse
• structural biology
• synaptic plasticity
• synaptic transmission

• Munc13
• calyx of Held
• phorbol ester
• posttetanic potentiation
• short-term plasticity

The readily releasable pool (RRP) of vesicles is a core concept in studies of presynaptic function. However, operating principles lack consensus definition and the utility for quantitative analysis has been questioned. Here we confirm that RRPs at calyces of Held from 14 to 21 day old mice have a fixed capacity for storing vesicles that is not modulated by Ca²⁺. Discrepancies with previous studies are explained by a dynamic flow-through pool, established during heavy use, containing vesicles that are released with low probability despite being immediately releasable. Quantitative analysis ruled out a posteriori explanations for the vesicles with low release probability, such as Ca²⁺-channel inactivation, and established unexpected boundary conditions for remaining alternatives. Vesicles in the flow-through pool could be incompletely primed, in which case the full sequence of priming steps downstream of recruitment to the RRP would have an average unitary rate of at least 9/s during heavy use. Alternatively, vesicles with low and high release probability could be recruited to distinct types of release sites; in this case the timing of recruitment would be similar at the two types, and the downstream transition from recruited to fully primed would be much faster. In either case, further analysis showed that activity accelerates the upstream step where vesicles are initially recruited to the RRP. Overall, our results show that the RRP can be well defined in the mathematical sense, and support the concept that the defining mechanism is a stable group of autonomous release sites.

Short-term plasticity has a dramatic impact on the connection strength of almost every type of synapse during normal use. Some synapses enhance, some depress, and many enhance or depress depending on the recent history of use. A better understanding is needed for modeling information processing in biological circuits and for studying the molecular biology of neurotransmission. Here we show that first principles at the calyx of Held, such as whether or not a readily-releasable pool of vesicles in the presynaptic terminal has a fixed capacity for storing vesicles, are unexpectedly similar to synapse types that are used at much lower frequencies. Our study establishes new methods for studying the function of presynaptic molecules, and the results suggest that a tractable general model of short-term plasticity can capture the full computational power of dynamic synaptic modulation across a large range of synapse types and situations.

• Animals
• Auditory Pathways/physiology
• Calcium/metabolism
• Cochlear Nucleus/physiology
• Computational Biology
• Computer Simulation
• Electric Stimulation
• Excitatory Postsynaptic Potentials
• Female
• Male
• Mice
• Mice, Inbred C57BL
• Models, Neurological
• Neuronal Plasticity
• Presynaptic Terminals/physiology
• Synaptic Vesicles/physiology
• Trapezoid Body/physiology

• P20 GM103554 NIGMS NIH HHS

• Calyx of Held
• Readily-releasable pool
• Release probability
• Synaptic transmission

Chromaffin cells release catecholamines by exocytosis, a process that includes vesicle docking, priming and fusion. Although all these steps have been intensively studied, some aspects of their mechanisms, particularly those regarding vesicle transport to the active sites situated at the membrane, are still unclear. In this work, we show that it is possible to extract information on vesicle motion in Chromaffin cells from the combination of Langevin simulations and amperometric measurements. We developed a numerical model based on Langevin simulations of vesicle motion towards the cell membrane and on the statistical analysis of vesicle arrival times. We also performed amperometric experiments in bovine-adrenal Chromaffin cells under Ba²⁺ stimulation to capture neurotransmitter releases during sustained exocytosis. In the sustained phase, each amperometric peak can be related to a single release from a new vesicle arriving at the active site. The amperometric signal can then be mapped into a spike-series of release events. We normalized the spike-series resulting from the current peaks using a time-rescaling transformation, thus making signals coming from different cells comparable. We discuss why the obtained spike-series may contain information about the motion of all vesicles leading to release of catecholamines. We show that the release statistics in our experiments considerably deviate from Poisson processes. Moreover, the interspike-time probability is reasonably well described by two-parameter gamma distributions. In order to interpret this result we computed the vesicles’ arrival statistics from our Langevin simulations. As expected, assuming purely diffusive vesicle motion we obtain Poisson statistics. However, if we assume that all vesicles are guided toward the membrane by an attractive harmonic potential, simulations also lead to gamma distributions of the interspike-time probability, in remarkably good agreement with experiment. We also show that including the fusion-time statistics in our model does not produce any significant changes on the results. These findings indicate that the motion of the whole ensemble of vesicles towards the membrane is directed and reflected in the amperometric signals. Our results confirm the conclusions of previous imaging studies performed on single vesicles that vesicles’ motion underneath plasma membranes is not purely random, but biased towards the membrane.

• Algorithms
• Animals
• Biological Transport
• Catecholamines/metabolism
• Cattle
• Chromaffin Cells/physiology
• Computer Simulation
• Electrophysiological Phenomena
• Exocytosis
• Models, Biological
• Models, Statistical
• Transport Vesicles/metabolism

• SNARE proteins
• asynchronous neurotransmission
• neurotransmitter release
• spontaneous neurotransmission
• synaptic transmission
• synaptic vesicle
• synchronous neurotransmission
• vesicle exocytosis
• vesicle pools

• MNTB
• calyx of Held
• complexin
• presynaptic mechanisms
• synaptic transmission
• synaptic vesicle fusion

• active zone
• calcium channels
• calyx of Held
• priming
• readily releasable pool
• synaptic vesicle

• amacrine cell
• bipolar cell
• release

• Animals
• Cell Line
• Mice
• Mice, Knockout
• Neurons/metabolism
• Qb-SNARE Proteins/genetics
• Qb-SNARE Proteins/metabolism
• Qc-SNARE Proteins/genetics
• Qc-SNARE Proteins/metabolism
• Synaptic Transmission/physiology
• Synaptotagmins/physiology

Action potentials trigger synchronous and asynchronous neurotransmitter release. Temporal properties of both types of release could be altered in an activity-dependent manner. While the effects of activity-dependent changes in synchronous release on postsynaptic signal integration have been studied, the contribution of asynchronous release to information transfer during natural stimulus patterns is unknown. Here we find that during trains of stimulations, asynchronous release contributes to the precision of action potential firing. Our data show that this form of release is selectively diminished in AP-3b2 KO animals, which lack functional neuronal AP-3, an adaptor protein regulating vesicle formation from endosomes generated during bulk endocytosis. We find that in the absence of neuronal AP-3, asynchronous release is attenuated and the activity-dependent increase in the precision of action potential timing is compromised. Lack of asynchronous release decreases the capacity of synaptic information transfer and renders synaptic communication less reliable in response to natural stimulus patterns.

• endocytosis
• synaptic transmission
• synaptic vesicle recruitment

• complexin
• neurotransmission
• synapses
• synaptotagmin
• vesicles
• voltage operated calcium channels