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. 2014 Mar 20;2(4):507–519. doi: 10.1016/j.stemcr.2014.01.018

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© 2014 The Authors

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License, which permits non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Immunohistochemical Characterization of Ventricular Ependymoglia Cells

(A) All ependymoglia cells are GFAP⁺; proliferation hot spots such as the dorsal pallium (Dp) and the bed nucleus of the stria terminalis (Bst) are marked as well as the striatum (Str). The drawing of the newt brain indicates the plane of section. Lines with arrows demarcate the borders of the hot spots. Arrows highlight even expression of GFAP in the hot spots.

(B) Sox2 and GFAP are expressed in all ependymoglia cells. Arrow highlights even expression of Sox2 in proliferating PCNA⁺ cells.

(C) GS/GFAP/MCM2 staining reveals type-1 and type-2 cells. Arrow points to MCM2⁺/GFAP⁺/GS⁻ (type-2) ependymoglia cells in hot spots. Arrowhead points to the surrounding nonproliferating GFAP⁺/GS⁺ (type-1) ependymoglia cells.

(D) Quantification of ependymoglia cells in hot spots coexpressing GFAP, GS, and MCM2. Note that most MCM2⁺ ependymoglia cells are type-2 cells (GFAP⁺/GS⁻). n = 5; p < 0.05.

(E) GS/GFAP/MCM2 expression in non-hot spot. Arrowhead points to proliferating type-1 cell (GFAP⁺/GS⁺).

(F) Quantification of ependymoglia cells in non-hot spots coexpressing GFAP, GS, and MCM2. Note that most MCM2⁺ ependymoglia cells are type-1 cells (GFAP⁺/GS⁺). n = 5; p < 0.05.

(G) Notch1⁻, type-2 ependymoglia cells (arrow) surrounded by Notch1⁺, type-1 ependymoglia cells (arrowhead). Note the colocalization of GS and Notch1 immunoreactivity.

(H) Quantification of GFAP-, Notch1-, and GS-expressing ependymoglia cells shows that, in hot spots, the vast majority of GFAP⁺ cells were type-1 (Notch1⁺/GS⁺). n = 4; p < 0.05.

(I) The majority of PCNA⁺ cells in hot spots are type-2 ependymoglia cells (GFAP⁺/Notch1⁻; arrow).

(J) Quantification of ependymoglia cells in hot spot regions coexpressing GFAP, Notch1, and PCNA. Note that almost all PCNA⁺ ependymoglia cells are type-2 cells (GFAP⁺/Notch1⁻). n = 5; p < 0.05.

(K and L) Few PCNA⁺ type-1 (GFAP⁺/Notch1⁺) ependymoglia cells were observed outside of hot spots. Quantified in (L). n = 5; p < 0.05.

(M) PSA-NCAM (NCAM) and type-2 (GFAP⁺/Notch1⁻) ependymoglia cells in hot spots (arrow). The surrounding region contains type-1 (GFAP⁺/Notch1⁺) ependymoglia cells and is devoid of PSA-NCAM expression (arrowhead).

(N) MCM2-expressing PSA-NCAM⁺ cells in hot spots (arrow).

Data represented as mean ± SEM. The scale bars represent 50 μm.