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. 2020 Jun 30;31(13):107838. doi: 10.1016/j.celrep.2020.107838

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© 2020 The Authors

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Neuronal Inactivation of Atrx in Atrx-cKO^Male Mice Causes Altered Hippocampal Morphology

(A and B) Immunofluorescence of the cortex (A) and hippocampus (B) of Ctrl^Male and Atrx-cKO^Male mice. Scale bar: 100 μm for cortex, 200 μm for hippocampus.

(C) Co-staining of ATRX (red) and NeuN (green) shows absence of ATRX in hippocampal neurons in Atrx-cKO^Male mice. CA1: cornu ammonis 1, CA2: cornu ammonis 2, CA3: cornu ammonis 3, DG: dentate gyrus. Scale bar: 200 μm.

(D) Western blot of ATRX in rostral cortex (RC), caudal cortex (CC), hippocampus (Hip), and cerebellum (Ceb) of Ctrl^Male and Atrx-cKO^Male mice. The fold change in protein expression normalized to INCENP (loading control) is indicated below.

(E) Atrx RNA transcripts were measured by qRT-PCR in the rostral cortex (Ctx) (p < 0.005) and hippocampus (Hip) (p < 0.001) (n = 4 of each genotype).

(F) Body weight of Ctrl^Male and Atrx-cKO^Male mice at 3 months of age (n = 10 each genotype; p < 0.05 by unpaired Student’s t test). MRI of 16 Ctrl^Male and 13 Atrx-cKO^Male fixed mouse brains.

(G) Absolute total volume of Ctrl^Male and Atrx-cKO^Male mouse brains (p < 0.0001).

(H) Coronal views of Ctrl^Male and Atrx-cKO^Male brains show changes in the density of absolute volume.

(I) Cumulative 3-dimensional (3D) image generated from Ctrl^Male and Atrx-cKO^Male mouse brains reveals altered hippocampal volume relative to brain size. Hippocampus is yellow, areas of increased volume in Atrx-cKO^Male are orange, and areas of decreased volume are green.

(J) Relative volume of CA1 stratum radiatum and lacunosum moleculare between Ctrl^Male and Atrx-cKO^Male mice (p < 0.05). MRI image at right shows cumulative changes in Atrx-cKO^Male compared to Ctrl^Male mice.

(K) Relative size of CA3 pyramidal layer between Ctrl^Male and Atrx-cKO^Male mice (p < 0.005). MRI image at right shows cumulative changes in Atrx-cKO^Male mice compared to controls.

(L) Relative volumes of all hippocampal structures, including mean and standard deviation of Ctrl^Male and Atrx-cKO^Male, percentage difference (%Diff), effect size, p value, and false discovery rate (FDR). Asterisks indicate p < 0.05.

See also Figure S1.