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. Author manuscript; available in PMC: 2010 Feb 12.
Published in final edited form as: Science. 2009 May 14;324(5934):1536–1540. doi: 10.1126/science.1173205

Figure 3. Synapse-, stimulus- and transcript-specific translation.

Figure 3

Sensorin translational reporter was expressed in SNs paired with MNs, SN soma was removed and dendra2 was UV photoconverted from green to red. Stimuli were locally applied to subsets of SN-MN synapses using perfusion electrodes (see fig S3). (A, B, C) Low magnification image of dendra2-reporter expressing coculture. Dashed circle indicates removed SN soma; blue arrowheads indicate direction of perfusion; black squares denote regions imaged before and after local perfusion; white squares denote imaged non-perfused regions. (A-C panels 1–4) Pseudocolored images of green dendra2 signal after photoconversion and before (pre) and after (post) local perfusion. (A-C panels 5 and 6) reporter mRNA FISH images. Arrowheads in A2 and A5 point to new translation colocalizing with reporter mRNA. A: 5’3’UTR reporter, local perfusion of 5x5HT. B: 5’3’UTR reporter, local perfusion of 5xFMRFa. C: 3’UTR, local perfusion of 5x5HT. See fig S6–9 for images of pre-UV green and post-UV red photoconverted dendra2 signal, 5XASW controls, and 1X5HT stimulation. Scale bar: 50µm; (D) Quantification of translation as ratio of ΔF/F at perfused compared to non-perfused sites reveals translation occurs only with 5x5HT. ***p<0.0001, Wilcoxon-Mann Whitney test (also see fig S10 for group data).