Fig. 3: ELS modifies astrocyte morphology in the LH. | Nature Communications

Fig. 3: ELS modifies astrocyte morphology in the LH.

From: A divergent astrocytic response to stress alters activity patterns via distinct mechanisms in male and female mice

Fig. 3

A Representative IHC of GFAP, Orexin- A, and Cx43, In the LH of Naïve and ELS mice, scale bars = 25 µm. B, C Normalised fluorescence intensity (Arb.U.) measures of GFAP and Cx43 in the LH of Naïve and ELS male mice. B GFAP - unpaired t test: Naïve = 80.33Arb.U., ±8.64 (N = 4, n = 20 cells), ELS = 55.91Arb.U., ±3.01 (N = 4, n = 20 cells), p = 0.011. C Cx43 - unpaired t test: Naïve= 93.74Arb.U., ±6.79 (N = 4, n = 20 cells), ELS = 72.68Arb.U., ±4.02 (N = 4, n = 20 cells), p = 0.011). D, E Normalised fluorescence intensity (Arb.U.) measures of GFAP and Cx43 in the LH of Naïve and ELS female mice. D GFAP - unpaired t test: Naïve =119.2Arb.U., ±12.7 (N = 4, n = 19 cells), ELS = 58.87Arb.U., ±3.91 (N = 4, n = 19 cells), p < 0.0001. E Cx43 - unpaired t test: Naïve = 100.8Arb.U., ±6.02 (N = 4, n = 20 cells), ELS = 79.27Arb.U., ±4.98 (N = 4, n = 20 cells), p = 0.009). F Quantification of Orexin-A+ cells between Naïve and ELS male mice (unpaired t test: Naïve = 17.33 cells, ±1.09 (N = 6), ELS = 16.83 cells, ±0.872 (N = 6), p = 0.727) and (G) Naïve and ELS female mice (unpaired t test: Naïve =15.60 cells, ±1.66 (N = 5), ELS = 15.71 cells, ±0.747 (N = 7), p = 0.933). H Quantification of S100β + astrocytes in the lateral hypothalamus (unpaired t test: Naïve = 14.50 (N = 8), ELS = 12.00 (N = 8), p = 0.367) scale bars = 25 µm. I Experimental timeline and viral constructs for sparse labelling of astrocytes in the LH. Scale bars = 50μm. J Representative AAV-GfaABC1D-eGFP expressing astrocytes in the LH from Naïve and ELS mice, scale bars = 5 μm. Morphological reconstruction of LH astrocytes from Naïve and ELS male mice. K Volume (unpaired t test: Naïve = 5916 μm3, ±2.797 (N = 4, n = 18 cells), ELS = 4430 μm3, ±308 (N = 3, n = 13 cells), p = 0.0004). L Total process length (unpaired t test: Naïve =3365 μm ±293 (N = 3, n = 12 cells), ELS = 3168μm, ±471(N = 3, n = 10 cells), p = 0.717). M No. of branch points (Mann–Whitney test: Naïve = 1256 branch points ±264 (N = 3, n = 12 cells), ELS = 943 branch points, ±263 (N = 3, n = 10 cells) p = 0.314). N Sholl analysis (two-way ANOVA; F(1,25) = 0.09, p = 0.764). Morphological reconstruction of LH astrocytes from Naïve and ELS female mice. O Volume (unpaired t test: Naïve = 7740 μm3, ±1041 (N = 4, n = 16 cells) ELS = 3659μm3, ±257 (N = 4, n = 19 cells), p = 0.0006). P Total process length (unpaired t test: Naïve =4396 μm, ±627 (N = 3, n = 10 cells), ELS = 2701 μm, ±278 (N = 4, n = 17 cells), p = 0.009). Q No. of branch points (Mann–Whitney test: Naïve = 1209 branch points, ±211 (N = 3, n = 11 cells), ELS = 720.5 branch points, ±278 (N = 4, n = 16 cells) p = 0.488). R Sholl analysis (two-way ANOVA; F(1,23 = 4.31, p = 0.049). N = mice, n = cells. Bar charts = mean ± S.E.M. ns = not significant, *P < 0.05, **P < 0.01.

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