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. 2017 Jan 18;93(2):409–424. doi: 10.1016/j.neuron.2016.11.046

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© 2017 The Author(s)

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Rising cAMP Levels Produce GluA3-Dependent Synaptic Potentiation without AMPAR Trafficking

(A) Left column is a Z_max projection of a stack of pictures showing a representative GluA3-SEP-transfected PC. In the top row, example pictures of a PC dendrite expressing GluA3-SEP before (middle) and after (right) FSK application were color-coded according to the fluorescence intensity to improve the visualization of, in this case, the absence of changes of surface GluA3-SEP over time. In the bottom row, example pictures of a PC dendrite expressing GluA3-SEP before (middle) and after (right) DHPG application reveal a significant reduction in synaptic GluA3-SEP over time. The right column shows that fluorescence intensity after FSK application, normalized by the fluorescence before application (FSK, middle bar), showed no significant increase of GuA3-SEP compared to the spines in which the drug was not applied (control, left bar). However, DHPG application significantly reduced GluA3-SEP in PC spines in accordance with the observed synaptic depression.

(B) Despite the lack of a detectable increase in surface GluA3-SEP, FSK produced a significant increase in mEPSC amplitude and frequency in GluA3-SEP-transfected PCs in organotypic slices. DHPG induced a significant decrease in mEPSC amplitude and frequency in these neurons.

(C) On the left is an example baseline maximum intensity projection z stack (3 μM, six optical planes) of a dendrite transfected with GluA3-SEP obtained with two-photon microscopy before, immediately after, and 30 min after photobleaching of the spine. The black traces above the pictures represent quantifications of SEP fluorescence across the spine and parallel to the dendrite. On the right is an overall quantification of spine FRAP dynamics over time for PCs transfected with GluA3-SEP, either with (n = 5) or without (n = 4) 50 μM FSK added after the moment of bleaching (0 min). SEP fluorescence intensity is normalized to baseline intensity (−5 min). No changes in SEP intensity were observed over time in spines neighboring the bleached spines.