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. 2019 Oct 28;8:e49493. doi: 10.7554/eLife.49493

Figure 2. Social isolation reduces vascular endothelin expression.

(A) Edn1 and Edn3 mRNA expression in laminin positive and CD31 positive blood vessels as revealed by RNAScope in situ hybridisation. (B) Representative images of Edn1 and Pecam1 mRNA expression in the mPFC. (C) Quantification of the number of Edn1 expressing Pecam1 positive endothelial cells. Grouped 0.8033 ± 0.04411 n = 4 mice, Isolated 0.5074 ± 0.05412 n = 5 mice (mean ± standard error). Mann-Whitney test, p=0.0159. (D) Quantification of the mean Edn1 mRNA molecules expressed by Pecam1 positive cells per mouse. Grouped 12.29 ± 3.312 n = 4 mice, Isolated 4.673 ± 0.4059 n = 5 mice (mean ± standard error). Mann-Whitney test, p=0.0159.

Figure 2.

Figure 2—figure supplement 1. EDN mRNA is not expressed in astrocytes and microglia.

Figure 2—figure supplement 1.

Expression of Edn1, Edn2 and Edn3 in S100β positive astrocytes and IBA1 positive microglia. (A) RNAScope in situ hybridisation for Edn1, Edn2 and Edn3 mRNA in the mouse medial prefrontal cortex stained for S100β positive astrocytes. Note that S100β positive cells are negative for Edn1 and Edn3 mRNA while positive Edn signal can be seen in S100β negative cells. (B) RNAScope in situ hybridisation for Edn1, Edn2 and Edn3 mRNA in the mouse medial prefrontal cortex stained for Iba1 positive microglia. Note that Iba1 positive cells are negative for Edn1 and Edn3 mRNA while positive Edn signal can be seen in Iba1 negative cells.
Figure 2—figure supplement 2. Social isolation does not affect medial prefrontal cortex vasculature.

Figure 2—figure supplement 2.

(A) Representative images of medial prefrontal cortex vasculature staining for PECAM1. (B) Quantification of PECAM1 area in medial prefrontal cortex layer II/III: Grouped 2.468% ± 0.3156 n = 6 mice, Isolated 2.086 ± 0.13 n = 6 mice (mean ± standard error). Mann-Whitney test. (C) Quantification of number of Pecam1 mRNA expressing cells per field of view. Grouped 9 ± 0.5888 n = 4 mice, Isolated 8.44 ± 1.162 n = 4 mice (mean ± standard error). Mann-Whitney test. (D) Quantification of the number of Pecam1 mRNA molecules. Grouped 13.47 ± 6.204 n = 77 cells from four mice, Isolated 10.89 ± 5.02 n = 92 cells from five mice (mean ± standard deviation). Mann-Whitney test, p=0.002. (E) Quantification of the number of Pecam1 mRNA molecules per mouse. Grouped 13.53 ± 1.305 n = 4 mice, Isolated 10.48 ± 1.018 n = 5 mice (mean ± standard error). Mann-Whitney test. (F) Quantification of the number of Edn1 mRNA molecules in Pecam1 positive endothelial cells. Grouped 12.08 ± 9.759 n = 77 cells from four mice, Isolated 4.837 ± 4.406 n = 92 from five mice (mean ± standard deviation). Mann-Whitney test, p<0.0001. (G) Quantification of the number of Edn3 expressing Pecam1 positive endothelial cells. Grouped 0.5463 ± 0.01586 n = 5 mice, Isolated 0.3674 ± 0.02163 n = 5 mice (mean ± standard error). Mann-Whitney test, p=0.0079.