Synaptic transmission between individual presynaptic terminals and postsynaptic dendrites is a fundamental element of communication among central nervous system neurons. Yet little is known about evoked neurotransmission at the level of single presynaptic boutons. Here we describe key functional characteristics of individual presynaptic boutons of hippocampal neurons in culture. Excitatory postsynaptic currents (e.p.s.cs) were evoked by localized application of elevated K+/Ca2+ solution to single functional boutons, visually identified by staining with the vital dye FM1-43 (refs 6, 7). Frequent repetitive stimulation produced a decline in the incidence of e.p.s.cs as the pool of releasable vesicles was exhausted; typically, recovery proceeded with a time constant of about 40 s (23 degrees C), and involved a vesicular pool capable of generating about 90 e.p.s.cs without recycling. At individual synapses, synaptic currents were broadly distributed in amplitude, but this distribution was remarkably similar at multiple synapses on a given postsynaptic neuron. The average size of synaptic currents and of responses to focal glutamate application varied fourfold across different cells, decreasing markedly with increasingly dense synaptic innervation. This raises the possibility of a very effective mechanism for coordinating synaptic strength at multiple sites throughout the dendritic tree.
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"The glutamate refilling time constant estimated here (15 s) is slower than that reported for vesicle acidification estimated using synapto-pHluorin (0.5–5 s) (Gandhi and Stevens, 2003), suggesting that intravesicular acidification precedes glutamate uptake in vesicles after endocytosis. This refilling time constant is faster than that of the recovery of EPSCs after depleting releasable vesicles by high-frequency stimulation (40 s, Liu and Tsien, 1995) and that of vesicles to become reavailable after KCl perfusion (30 s, Ryan et al., 1993), suggesting that most vesicles are fully refilled during recycling. In vesicle recycling steps, fast and slow endocytosis ranging from subseconds to tens of "
[Show abstract][Hide abstract] ABSTRACT: After releasing neurotransmitter, synaptic vesicles are retrieved by endocytosis and recycled via fast and slow pathways to be reused for synaptic transmission. To maintain the synaptic efficacy, vesicles must be refilled with neurotransmitter during recycling. However, the refilling speed estimated in isolated or reconstructed vesicles is, thus far, too slow to fill up vesicles within the period of recycling. Here, we re-examined the vesicle refilling rate directly at central glutamatergic synapses in slices, using simultaneous presynaptic and postsynaptic whole-cell recording combined with caged glutamate photolysis. After washing out vesicular glutamate, refilling of vesicles with uncaged glutamate caused a recovery of EPSCs with a time constant of 15 s that varied depending upon temperature, age, and cytosolic Cl(-) concentrations. This time constant is faster than that of the slow recycling pathway (∼30 s) after clathrin-mediated endocytosis but is much too slow to fill up vesicles replenished through fast recycling pathways (<1 s).
"Calibration of synaptic strength to higher level    via constitutive insertion of somatically synthesized GluA1/2 AMPARs  Established network: Sudden decrease in output with concurrent decrease in presynaptic inputs "
[Show abstract][Hide abstract] ABSTRACT: Homeostatic plasticity has emerged as a fundamental regulatory principle that strives to maintain neuronal activity within optimal ranges by altering diverse aspects of neuronal function. Adaptation to network activity is often viewed as an essential negative feedback restraint that prevents runaway excitation or inhibition. However, the precise importance of these homeostatic functions is often theoretical rather than empirically derived. Moreover, a remarkable multiplicity of homeostatic adaptations has been observed. To clarify these issues, it may prove useful to ask: why do homeostatic mechanisms exist, what advantages do these adaptive responses confer on a given cell population, and why are there so many seemingly divergent effects? Here, we approach these questions by applying the principles of control theory to homeostatic synaptic plasticity of mammalian neurons and suggest that the varied responses observed may represent distinct functional classes of control mechanisms directed toward disparate physiological goals.
"In addition, their ACSF included GABA B R and GABA A R blockers, and [Ca 2+ ] o twice as high as in our study, making the direct comparison problematic. Local fine-tuning of presynaptic GABA B R tone may be involved in synaptic scaling along dendritic compartment, leading to homeostasis of basal synaptic activity through a negative correlation between the number of functional boutons and unitary synaptic strength (Liu and Tsien, 1995) or its presynaptic determinant, Pr (Branco et al., 2008 "
[Show abstract][Hide abstract] ABSTRACT: Presynaptic GABA(B) receptor (GABA(B)R) heterodimers are composed of GB(1a)/GB(2) subunits and critically influence synaptic and cognitive functions. Here, we explored local GABA(B)R activation by integrating optical tools for monitoring receptor conformation and synaptic vesicle release at individual presynaptic boutons of hippocampal neurons. Utilizing fluorescence resonance energy transfer (FRET) spectroscopy, we detected a wide range of FRET values for CFP/YFP-tagged GB(1a)/GB(2) receptors that negatively correlated with release probabilities at single synapses. High FRET of GABA(B)Rs associated with low release probability. Notably, pharmacological manipulations that either reduced or increased basal receptor activation decreased intersynapse variability of GB(1a)/GB(2) receptor conformation. Despite variability along axons, presynaptic GABA(B)R tone was dendrite specific, having a greater impact on synapses at highly innervated proximal branches. Prolonged neuronal inactivity reduced basal receptor activation, leading to homeostatic augmentation of release probability. Our findings suggest that local variations in basal GABA concentration are a major determinant of GB(1a)/GB(2) conformational variability, which contributes to heterogeneity of neurotransmitter release at hippocampal synapses.