title: Counting numbers of synaptic proteins: absolute quantification and single molecule imaging techniques.
abstract: The ability to count molecules is essential to elucidating cellular mechanisms, as these often depend on the absolute numbers and concentrations of molecules within specific compartments. Such is the case at chemical synapses, where the transmission of information from presynaptic to postsynaptic terminals requires complex interactions between small sets of molecules. Be it the subunit stoichiometry specifying neurotransmitter receptor properties, the copy numbers of scaffold proteins setting the limit of receptor accumulation at synapses, or protein packing densities shaping the molecular organization and plasticity of the postsynaptic density, all of these depend on exact quantities of components. A variety of proteomic, electrophysiological, and quantitative imaging techniques have yielded insights into the molecular composition of synaptic complexes. In this review, we compare the different quantitative approaches and consider the potential of single molecule imaging techniques for the quantification of synaptic components. We also discuss specific neurobiological data to contextualize the obtained numbers and to explain how they aid our understanding of synaptic structure and function.
authors.affiliation: Institute of Biology , Biologie Cellulaire de la Synapse, Inserm U1024, CNRS 8197, École Normale Supérieure (ENS), 46 rue d'Ulm, Paris 75005, France.
authors.forename: Angela
authors.initials: A
authors.lastname: Patrizio
authors.affiliation: Institute of Biology , Biologie Cellulaire de la Synapse, Inserm U1024, CNRS 8197, École Normale Supérieure (ENS), 46 rue d'Ulm, Paris 75005, France.
authors.forename: Christian G
authors.initials: CG
authors.lastname: Specht
keywords: pair correlation analysis; photoactivated localization microscopy; single molecule localization microscopy; stochastic optical reconstruction microscopy