RT Journal Article SR Electronic T1 Molecular Diversity of Glutamatergic and GABAergic Synapses from Multiplexed Fluorescence Imaging JF eneuro JO eNeuro FD Society for Neuroscience SP ENEURO.0286-20.2020 DO 10.1523/ENEURO.0286-20.2020 VO 8 IS 1 A1 Eric Danielson A1 Karen Perez de Arce A1 Beth Cimini A1 Eike-Christian Wamhoff A1 Shantanu Singh A1 Jeffrey R. Cottrell A1 Anne E. Carpenter A1 Mark Bathe YR 2021 UL http://www.eneuro.org/content/8/1/ENEURO.0286-20.2020.abstract AB Neuronal synapses contain hundreds of different protein species important for regulating signal transmission. Characterizing differential expression profiles of proteins within synapses in distinct regions of the brain has revealed a high degree of synaptic diversity defined by unique molecular organization. Multiplexed imaging of in vitro rat primary hippocampal culture models at single synapse resolution offers new opportunities for exploring synaptic reorganization in response to chemical and genetic perturbations. Here, we combine 12-color multiplexed fluorescence imaging with quantitative image analysis and machine learning to identify novel synaptic subtypes within excitatory and inhibitory synapses based on the expression profiles of major synaptic components. We characterize differences in the correlated expression of proteins within these subtypes and we examine how the distribution of these synapses is modified following induction of synaptic plasticity. Under chronic suppression of neuronal activity, phenotypic characterization revealed coordinated increases in both excitatory and inhibitory protein levels without changes in the distribution of synaptic subtypes, suggesting concerted events targeting glutamatergic and GABAergic synapses. Our results offer molecular insight into the mechanisms of synaptic plasticity.