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. 2020 Jan 7;11:72. doi: 10.1038/s41467-019-13809-8

Fig. 5. GluN2B expression in SST interneurons mediates effect of ketamine on dendritic spine calcium.

Fig. 5

a Schematic of AAV1-CMV-dsRed-pSico-GluN2BshRNA (standard AAV cassettes not drawn) for Cre-dependent expression of shRNA against GluN2B. b Top, timeline for experiments involving selective knockdown of GluN2B expression in SST interneurons (GluN2B-SST KD). Bottom, in vivo two-photon image of ubiquitous dsRed expression (left), Cre-dependent GCaMP6s expression (middle), and a composite image (right). Arrowheads, cells that expressed both dsRed and GCaMP6s. c The rate of spontaneous calcium events for GCaMP6s-expressing SST interneuron cell bodies with or without GluN2B-SST KD (GluN2B-SST KD: 1.5 ± 0.2 Hz, control SST-IRES-Cre animals with no KD: 1.9 ± 0.1 Hz, mean ± s.e.m.; P = 0.04, two-sample t-test). For GluN2B-SST KD, n = 58 cells from 3 animals. For control animals, n = 72 cells from 3 animals. d The normalized difference in the number of spontaneous calcium events GCaMP6s-expressing SST interneuron cell bodies, for GluN2B-SST KD with ketamine or saline injection. Normalized difference was calculated as post-injection minus pre-injection values normalized by the pre-injection value (ketamine (10 mg/kg) with GluN2B-SST KD: 2 ± 4%, saline with GluN2B-SST KD: 7 ± 6%, mean ± s.e.m.; P = 0.46, two-sample t-test). For ketamine, n = 58 cells from 3 animals. For saline, n = 39 cells from 3 animals. e Same as (c) for GCaMP6f-expressing apical dendritic spines (GluN2B-SST KD: 0.85 ± 0.03 Hz, mean ± s.e.m., control SST-IRES-Cre animals: 0.73 ± 0.04 Hz, mean ± s.e.m.; P = 0.01, two-sample t-test). For GluN2B-SST KD, n = 277 spines from 4 animals. For control animals, n = 118 spines from 3 animals. f Same as (d) for GCaMP6f-expressing apical dendritic spines (ketamine (10 mg/kg) with GluN2B-SST KD: −4 ± 1%, saline with GluN2B-SST KD: −5 ± 2%, mean ± s.e.m.; P = 0.8, two-sample t-test). For ketamine, n = 169 spines from 4 animals. For saline, n = 171 spines from 4 animals. *P < 0.05; **P < 0.01; ***P < 0.001; n.s., not significant.