Hafner, A. S., Donlin-Asp, P. G., Leitch, B., Herzog, E., & Schuman, E. M. (2019). Local
protein synthesis is a ubiquitous feature of neuronal pre-and postsynaptic compartments. Science,
364(6441), eaau3644. https://science.sciencemag.org/content/364/6441/eaau3644.abstract
“Efforts to localize molecules or cell biological events to neuronal pre- or
postsynaptic compartments by using fluorescence microscopy are limited by the tight
association of the axonal bouton and the dendritic spine; the synaptic cleft, the only clear region
of separation, is only ~20 nm wide. In this study, to increase the resolving power to visualize
RNA molecules in pre- and postsynaptic compartments, we optimized fluorescence in situ
hybridization (FISH) and nascent protein detection methods for use with expansion microscopy.
… To obtain direct evidence for protein synthesis in synaptic compartments, particularly
presynaptic boutons, we adapted the puromycin-based metabolic labeling strategy for detection
with electron or expansion microscopy.”
Mouse cortex and hippocampal culture
A range of advanced labeling and microscopy techniques are used to dissect the
regulation of local protein synthesis pre- and postsynaptically in response to plasticity induction.
“In adult rodent brain slices and cultured neurons, we found that >75% of both excitatory
and inhibitory presynaptic terminals contain the machinery for protein synthesis: rRNA,
ribosomes, and polyadenylated [poly (A)+] mRNA.”