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. 2008 Apr 23;28(17):4336–4349. doi: 10.1523/JNEUROSCI.4379-07.2008

Figure 1.

Figure 1.

Characterization of transfected sst2A receptors in hippocampal neurons. A, Hippocampal neurons were transfected with a plasmid encoding the wild-type receptor (sst2A-WT), fixed, and processed for fluorescence immunocytochemistry for visualization of the receptor by confocal microscopy. Note the intense sst2A-WT fluorescence localized at the membrane of the soma and the dendrites of this transfected neuron. sst2A-WT fluorescence is also visible in spine heads. Scale bar, 25 μm. B, C, Hippocampal neurons were transfected with a plasmid encoding the sst2A receptor tagged with EGFP (sst2A-EGFP) and processed for live-cell confocal microscopy. As observed for the wild-type receptor, the sst2A-EGFP is enriched at the somatodendritic membrane domain as well as in spine heads. Scale bars: B, 50 μm; C, 5 μm. D, Expression of endogenous and transfected sst2A receptors as measured by immunocytochemistry and confocal microscopy. The bar charts represent mean fluorescence measured at the membrane of neurons expressing the endogenous receptor or expressing the sst2A-WT or the sst2A-EGFP receptor (n = 10; mean ± SEM; **p < 0.001; a.u., arbitrary unit). E, CHO cells were transfected with pCRE-SEAP alone or cotransfected with pCRE-SEAP and either the sst2A-WT or sst2A-EGFP receptor. OCT induces a significant decrease in the level of cAMP in CHO cells stimulated by forskolin (Fsk) and expressing the sst2A-WT or sst2A-EGFP receptor. This effect was not observed in cells expressing pCRE-SEAP alone (n = 6; mean ± SEM; **p < 0.001, ***p < 0.0001; ns, nonsignificant; a.u., arbitrary unit). F, Representative time series from a FRAP experiment in spine heads. Bleaching of sst2A-EGFP (delimited by black borders) is followed by rapid recovery of fluorescence. Scale bar, 2 μm. G, Graph shows normalized FRAP curve from bleached area shown in F. H, Averaged mobile fractions, half-time of recoveries (T half), and diffusion coefficients for somatic membrane, dendritic membrane, and spine heads (n = 15; mean ± SEM; *p < 0.05, **p < 0.001).