title: Counting Vesicular Release Events Reveals Binomial Release Statistics at Single Glutamatergic Synapses.
abstract: Many central glutamatergic synapses contain a single presynaptic active zone and a single postsynaptic density. However, the basic functional properties of such "simple synapses" remain unclear. One important step toward understanding simple synapse function is to analyze the number of synaptic vesicles released in such structures per action potential, but this goal has remained elusive until now. Here, we describe procedures that allow reliable vesicular release counting at simple synapses between parallel fibers and molecular layer interneurons of rat cerebellar slices. Our analysis involves local extracellular stimulation of single parallel fibers and deconvolution of resulting EPSCs using quantal signals as template. We observed a reduction of quantal amplitudes (amplitude occlusion) in pairs of consecutive EPSCs due to receptor saturation. This effect is larger (62%) than previously reported and primarily reflects receptor activation rather than desensitization. In addition to activation-driven amplitude occlusion, each EPSC reduces amplitudes of subsequent events by an estimated 3% due to cumulative desensitization. Vesicular release counts at simple synapses follow binomial statistics with a maximum that varies from 2 to 10 among experiments. This maximum presumably reflects the number of docking sites at a given synapse. These results show striking similarities, as well as significant quantitative differences, with respect to previous results at simple GABAergic synapses.
  
  
  CONCLUSIONS
  It is generally accepted that the output signal of individual central synapses saturates at high release probability, but it remains unclear whether the source of saturation is presynaptic, postsynaptic, or both presynaptic and postsynaptic. To clarify this and other issues concerning the function of synapses, we have developed new recording and analysis methods at single central glutamatergic synapses. We find that individual release events engage a high proportion of postsynaptic receptors (62%), revealing a larger component of postsynaptic saturation than anticipated. Conversely, we also find that the number of released synaptic vesicles is limited at each active zone. Altogether, our results argue for both presynaptic and postsynaptic contributions to signal saturation at single glutamatergic synapses.
authors.affiliation: Laboratoire de Physiologie Cérébrale, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8118, Université Paris Descartes, 75006 Paris, France, and.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Malagon
authors.affiliation: Laboratoire de Physiologie Cérébrale, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8118, Université Paris Descartes, 75006 Paris, France, and.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Miki
authors.affiliation: Laboratoire de Physiologie Cérébrale, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8118, Université Paris Descartes, 75006 Paris, France, and.
authors.forename: Isabel
authors.initials: I
authors.lastname: Llano
authors.affiliation: Max Planck Institute for Biophysical Chemistry, D37077 Göttingen, Germany.
authors.forename: Erwin
authors.identifier: http://orcid.org/0000-0002-9758-7922
authors.initials: E
authors.lastname: Neher
authors.affiliation: Laboratoire de Physiologie Cérébrale, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8118, Université Paris Descartes, 75006 Paris, France, and alain.marty@parisdescartes.fr.
authors.forename: Alain
authors.initials: A
authors.lastname: Marty
mesh_terms: D000465:Algorithms; D000818:Animals; D002531:Cerebellum / Q000502:physiology; D004558:Electric Stimulation; D019706:Excitatory Postsynaptic Potentials / Q000502:physiology; D005260:Female; D018698:Glutamic Acid / Q000502:physiology*; D007395:Interneurons / Q000502:physiology; D008297:Male; D008959:Models, Neurological; D051381:Rats; D017207:Rats, Sprague-Dawley; D018091:Receptors, AMPA / Q000502:physiology*; D013569:Synapses / Q000378:metabolism* / Q000502:physiology*; D009435:Synaptic Transmission / Q000502:physiology; D013572:Synaptic Vesicles / Q000502:physiology
chemical_list: D018091:Receptors, AMPA; D018698:Glutamic Acid
keywords: AMPA receptor; cerebellum; glutamate; synaptic release; synaptic vesicles