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. 2021 Sep 28;8(5):ENEURO.0152-21.2021. doi: 10.1523/ENEURO.0152-21.2021

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Copyright © 2021 Gould et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 6. — Reduced QR2 expression in SST interneurons enhances novel taste memory formation. A, Schematic diagram of pAAV-Sico-Red-QR2 shRNA vector before and after Cre-mediated recombination. B, Representative image of Cre-dependent QR2 shRNA viral vector expression in GAD Cre mouse aIC. C, Mice were injected with the Cre-dependent QR2 shRNA or scrambled viral vector to the aIC. A month later, they underwent novel taste learning, and 2 d after consuming the novel taste, they were given a choice test and a preference index was made. A week later, the mice were killed to assess infection efficacy, and the effect the expression of the virus had on local QR2 mRNA expression. D, GAD Cre mice injected with Cre-dependent QR2 shRNA showed significantly improved novel taste memory compared with scrambled controls (GAD Cre-Scrambled 43.68 ± 7.218%, n = 9; GAD Cre-QR2 shRNA 67.31 ± 5.096%; n = 10; Student’s t test, t = 2.717 df = 17, p = 0.0146). E, GAD Cre mice injected with Cre-dependent QR2 shRNA showed significantly reduced QR2 mRNA expression compared with scrambled controls (GAD Cre-Scrambled 1.007 ± 0.066 2^-ΔΔCt, n = 4; GAD Cre-QR2 shRNA 0.813 ± 0.019 2^-ΔΔCt, n = 5; Mann–Whitney test, p = 0.0159). F, Mice co-expressing Cre under the CamKII promoter and Cre-dependent QR2 shRNA in the aIC show no significant change in novel taste memory compared with those injected with the scrambled Cre-dependent virus (CamKII Cre-Scrambled 56.48 ± 8.423%, n = 10; CamKII Cre-QR2 shRNA 42.84 ± 8.324%, n = 8; Student’s t test, t = 1.136 df = 16, p = 0.2728). G, A significant reduction in QR2 mRNA was measured in mice co-expressing Cre under CamKII promoter and Cre-dependent QR2 shRNA in the aIC, compared with controls (CamKII Cre-Scrambled 1.027 ± 0.119 2^-ΔΔCt, n = 5; CamKII Cre-QR2 shRNA 0.7122 ± 0.049 2^-ΔΔCt, n = 5; Student’s t test, t = 2.435 df = 8, p = 0.0409). H, PV Cre mice injected with Cre-dependent QR2 shRNA in the aIC show no significant change in novel taste memory compared with those injected with the scrambled Cre-dependent virus (PV Cre-Scrambled 56.56 ± 6.274%, n = 7; PV Cre-QR2 shRNA 54.67 ± 8.180%, n = 8; Student’s t test, t = 0.1792 df = 13, p = 0.8605). I, PV Cre mice injected with Cre-dependent QR2 shRNA showed insignificant QR2 mRNA expression reduction compared with scrambled controls (PV Cre-Scrambled 1.003 ± 0.035 2^-ΔΔCt, n = 5; PV Cre-QR2 shRNA 0.940 ± 0.056 2^-ΔΔCt, n = 6; Student’s t test, t = 0.8789 df = 9, p = 0.4023). J, SST Cre mice injected with Cre-dependent QR2 shRNA showed significantly improved novel taste memory compared with scrambled controls (SST Cre-Scrambled 52.26 ± 7.868%, n = 6; SST Cre-QR2 shRNA 74.26 ± 5.843%; n = 6; Student’s t test, t = 2.246 df = 10, p = 0.0485). K, SST Cre mice injected with Cre-dependent QR2 shRNA showed insignificant QR2 mRNA expression reduction compared with scrambled controls (SST Cre-Scrambled 1.004 ± 0.046 2^-ΔΔCt, n = 5; SST Cre-QR2 shRNA 0.935 ± 0.032 2^-ΔΔCt, n = 4; Student’s t test, t = 1.154 df = 7, p = 0.2864). Data are shown as mean ± SEM; *p < 0.05.