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. 2015 Apr 23;6:6887. doi: 10.1038/ncomms7887

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(a–h) Triple immunochemistry for Nkx2.1, Ki67 and Nestin on E15 coronal slices from Nkx2.1-Cre⁻/Rosa-DTA (n=3; a–d) and Nkx2.1-Cre⁺/Rosa-DTA (n=3; e–h) mice. (i–p) Double immunochemistry for Nkx2.1 and GLAST on E16.5 (i–l) and for Olig2 and GLAST on E18.5 (m–p) coronal slices from Nkx2.1-Cre⁻/Rosa-DTA (i,j,m,n) and Nkx2.1-Cre⁺/Rosa-DTA (k,l,o,p) mice. (b–d,f–h,j,l,n,p) High-magnified views of the regions indicated by an arrow in a,e,i,k,m and o. (a–l) The GLAST⁺/Nestin⁺/Nkx2.1⁺/Ki67⁻ post-mitotic tunnel astroglia forming a palisade (*) surrounding the AC are depleted in the Nkx2.1-Cre⁺/Rosa-DTA (e–h,k,l) compared with control (a–d,i,j) mice. (m,n) In control mice, numerous GLAST⁺/Olig2⁻, GLAST⁻/Olig2⁺ and GLAST⁺/Olig2⁺ glial cells are observed within and around the AC. (o,p) In Nkx2.1-Cre⁺/Rosa-DTA mice, the number of AC GLAST⁺/Olig2⁻, GLAST⁻/Olig2⁺ and GLAST⁺/Olig2⁺ is decreased after cell ablation. Cell nuclei were counterstained in blue with Hoechst (a,b,e,f). The scale bar shown in panel a corresponds to the following length for panel(s) specified in brackets: 675 μm in a,e,i,k,m,o; 160 μm (b,c,f,g,j,l,n,p); 50 μm (d,h). (q) Bars (means±s.e.m.) from Nkx2.1-Cre⁻/Rosa-DTA (n=5) and Nkx2.1-Cre⁺/Rosa-DTA (n=3) brains represent the cell densities of GLAST⁺/Nkx2.1⁺ at the AC midline at E16.5. After quantification, the density of GLAST⁺ astrocytes expressing Nkx2.1 was decreased in Nkx2.1-Cre⁺/Rosa-DTA mice compared with control mice. (r) Bars (means±s.e.m.) from Nkx2.1-Cre⁻/Rosa-DTA (n=5) and Nkx2.1-Cre⁺/Rosa-DTA (n=5) brains represent the cell densities of GLAST⁺/Olig2⁻, GLAST⁻/Olig2⁺ and GLAST⁺/Olig2⁺ glial cells at the AC midline at E18.5. After quantification, the density of GLAST⁺ astrocytes expressing or not Olig2 was significantly decreased in Nkx2.1-Cre⁺/Rosa-DTA mice compared with control mice (**P<0.01, n=5, Student's t-test).