Xenbase Image ID: 124849

Figure 6. Ago2 expression enhances RNAi reduction of AMPA receptor-mediated synaptic responses in vivo. Whole-cell recordings were made from CNS neurons of Xenopus tadpoles transfected with pSi construct expressing shRNAs targeting the GluR2 subunit of AMPA receptor with or without Ago2 co-expression. (A) Diagram of the pSi constructs containing EnU6-shGluR2s for GluR2 knockdown, Ago2 for RNAi enhancement and GFP for cell identification. (B) Representative traces of spontaneous AMPA mEPSCs, superimposed from 30 consecutive traces. Cells co-expressing Ago2 with shGluR2-1 or shGluR2-2 (red) appear to have fewer mEPSCs than cells expressing shGluR2-1 or shGluR2-2 alone (black) and other control (green) traces. (C) Frequency of AMPA mEPSC in neurons transfected with pSi construct expressing shGluR2s with Ago2 (red) is significantly less than cells transfected with the pSi construct expressing shScrambled + Ago2 or GFP alone ((green) and neurons expressing shGluRs only (black) (p < 0.05, ANOVA). Neurons expressing shGluR2s without Ago2 co-expression (black) have mEPSC frequency comparable to controls transfected with shScrambled + Ago2 and GFP alone (green). Neurons expressing GFP alone, Ago2 alone and shScrambled + Ago2 have comparable AMPA mEPSC frequencies. Dashed gray line indicates the averaged frequency of shScrambled control. (D) Amplitudes of AMPA mEPSC are not different between neurons transfected with GFP alone, Ago2 alone, and shScrambled or shGluR2s with or without Ago2 co-expression. Dashed gray line indicates the averaged amplitude of shScrambled control. Image published in: Chen CM et al. (2009) Image downloaded from an Open Access article in PubMed Central. Copyright © 2009 Chen, Chiu, Shen and Cline. Larger Image Printer Friendly View

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