Fig. 4: The IBIST-based Ca2+ recording for cells defined by genetic marker and neural connectivity.
From: An intein-split transactivator for intersectional neural imaging and optogenetic manipulation
a Scheme illustrating AAV injections to define cells by two features: anterograde transsynaptic tracing and CaMKIIα-specific fluorophore labeling. b Examples showing fluorescence of the ventral hippocampus infected by AAVs of Flpo, tTAN (magenta), tTAC and TRE-GCaMP6s (GCa., green) and co-injected with blue beads (magenta). Scale bars: 500 µm. c, Summary of fluorescent intensity of GCaMP6s in b. One-way ANOVA between groups: F(2,33) = 66.19, ****P = 2.82 × 10−12. Turkey’s multiple comparisons: tTAN vs. tTAN+tTAC, ****P = 4.91 × 10−11; tTAC vs. tTAN+tTAC, ****P = 3.88 × 10−11. N = 3 animals/group. d Scheme illustrating the open field test. e Box plot of the percentage time in the center zone and surround. Center vs. surround, 8.2 ± 1.0% vs. 91.8 ± 1.0%, paired t-test, ****P = 2.08 × 10−18; N = 17 animals. f Examples of averaged traces of Ca2+ signals and animal speeds upon center entry (green) and exit (black) in the open field test. g Summary of normalized Ca2+ responses upon center entry and exit. One-way ANOVA between groups (tTAN, 0.3 ± 0.2%, N = 6 animals; tTAC, 0.4 ± 0.2%, N = 7 animals; tTAN+tTAC, 1.7 ± 0.8%, N = 4 animals; F(2,14) = 4.275, *P = 0.0356; Turkey’s multiple comparisons, *P < 0.05). h Scheme illustrating the foot shock test. i Examples of heatmaps and averaged traces of Ca2+ signals upon foot shocks (between dash lines,). j, Summary of Ca2+ responses. One-way ANOVA between groups in first session (tTAN, 0.1 ± 0.2%, N = 6 animals; tTAC, 0.01 ± 0.01%, N = 7 animals; tTAN+tTAC, 3.9 ± 1.1%, N = 4 animals; F(2,14) = 21.17, ****P = 5.7 × 10−5; Turkey’s multiple comparisons, ***P < 0.001) and second session (tTAN, 0.1 ± 0.1%, N = 6 animals; tTAC, 0.5 ± 0.2%, N = 7 animals; tTAN+tTAC, 4.3 ± 0.2%, N = 4 animals; F(2,14) = 141.3, ****P = 5.23 × 10−10; Turkey’s multiple comparisons, ****P < 0.0001). Data summary: mean ± SEM. Statistical analysis: two sided. Box plots: whiskers (min/max), middle line (median), plus (mean), box (25/75 percentile). Source data are provided as a Source Data file.
