Extended Data Fig. 7: Validation of in vivo two-photon calcium imaging of binocular zone neurons.
From: Astrocyte CCN1 stabilizes neural circuits in the adult brain
a. Retinotopic mapping of the BZ with small checkerboard boxes. b. Representative retinotopy in degrees of visual field in example mouse “L022”. Top, horizontal retinotopy, “azimuth”. Bottom, vertical retinotopy, “elevation”. Left, contralateral eye responses. Right, ipsilateral eye responses. Scale bar, 100 µm. c. Example mouse “L022”. Histograms of number of cells with peak responses to a particular azimuth (horizontal) or elevation (vertical). d. Validation of longitudinal tracking. Example mouse “L022”. Mean images pre, left, and post, right, MD. e. Left, fused image. Right, white denotes overlapping ROIs. f. Percent of longitudinally tracked cells across WT and CCN1 cKO mice. g. Unresponsive cells as a proportion of total cells. Corrected chi-square tests (4 comparisons; α = 0.0125). Stars, significance. h. Change in unresponsive cell proportion after MD, data and statistics from g. Stars, significant change relative to pre MD. i. C2I, contralateral to ipsilateral. C2B, contralateral to binocular. C2U, contralateral to unresponsive. j. B2M, binocular to monocular. M2B, monocular to binocular. Stable B, stable binocular. k. C2I, contralateral to ipsilateral. B2I, binocular to ipsilateral. U2I, unresponsive to ipsilateral. l. I2U, ipsilateral to unresponsive. C2U, contralateral to unresponsive. B2U, binocular to unresponsive. i-l. Corrected chi-square tests (3 comparisons; α = 0.01667). Stars, significance. WT total longitudinally tracked cells n = 453. CCN1 cKO total longitudinally tracked cells n = 511. m. ODI of longitudinally imaged responsive neurons only. WT n = 229 cells. cKO. n = 163 cells. n. Binocular matching as difference in preferred orientation for contralateral and ipsilateral responses for longitudinally tracked binocular neurons. WT n = 73 cells. cKO n = 37 cells. 4 mice/ genotype. m and n. Kruskal-Wallis tests with post-hoc Dunn’s tests on cells. For all dot plots, dots are individual cells. Lines and bars, median ± 95% CI. o. Mouse average peak ΔF/F for contralateral and ipsilateral. Two-way ANOVAs with paired uncorrected Fisher’s LSD tests on mouse averages. 4 comparisons; α = 0.0125. Starred p-values, significance. Bar graphs, mean ± SEM.
