Figure 2

We transfected rat neuronal cultures with tdTomato to visualize spine morphology. On the day of the experiment, neurons were incubated in a buffer containing MNI-caged-L-glutamate and DQ™ gelatin, a fluorogenic gelatin, which is a canonical substrate for MMP-9. DQ-gelatin consists of highly quenched, fluorescein-labeled gelatin, which upon proteolytic digestion emits green fluorescence, indicating local enzymatic activity of MMP-9. We stimulated single dendritic spines of tdTomato-expressing neurons by glutamate uncaging using structural LTP (sLTP)-inducing protocol. We measured the structural plasticity in the tdTomato (red) fluorescence channel and DQ™ gelatin digestion in the green channel. The file contains fluorescence changes for stimulated and nearby spines in both channels for every analysed spine.