4.2 Article

SynapsEM: Computer-Assisted Synapse Morphometry

Journal

FRONTIERS IN SYNAPTIC NEUROSCIENCE
Volume 12, Issue -, Pages -

Publisher

FRONTIERS RESEARCH FOUNDATION
DOI: 10.3389/fnsyn.2020.584549

Keywords

electron microscopy; synapse; morphometry; ultrastructural analysis; SynapsEM

Categories

Funding

  1. Johns Hopkins University School of Medicine
  2. Johns Hopkins Discovery funds
  3. Johns Hopkins Catalyst Award
  4. National Science Foundation [1727260, 2016217537]
  5. National Institutes of Health [1DP2 NS111133-01, 1R01 NS105810-01A1, T32 GM007445, R01 NS034307]
  6. Directorate For Engineering
  7. Div Of Civil, Mechanical, & Manufact Inn [1727260] Funding Source: National Science Foundation

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The structural features of a synapse help determine its function. Synapses are extremely small and tightly packed with vesicles and other organelles. Visualizing synaptic structure requires imaging by electron microscopy, and the features in micrographs must be quantified, a process called morphometry. Three parameters are typically assessed from each specimen: (1) the sizes of individual vesicles and organelles; (2) the absolute number and densities of organelles; and (3) distances between organelles and key features at synapses, such as active zone membranes and dense projections. For data to be meaningful, the analysis must be repeated from hundreds to thousands of images from several biological replicates, a daunting task. Here we report a custom computer program to analyze key structural features of synapses: SynapsEM. In short, we developed ImageJ/Fiji macros to record x,y-coordinates of segmented structures. The coordinates are then exported as text files. Independent investigators can reload the images and text files to reexamine the segmentation using ImageJ. The Matlab program then calculates and reports key synaptic parameters from the coordinates. Since the values are calculated from coordinates, rather than measured from each micrograph, other parameters such as locations of docked vesicles relative to the center of an active zone can be extracted in Matlab by additional scripting. Thus, this program can accelerate the morphometry of synapses and promote a more comprehensive analysis of synaptic ultrastructure.

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