Nuclei susceptible to apoptosis in TC mutants exhibited the combinatorial expression of dominant γC4 and stochastic isoforms
(A) A coronal section of the midbrain of an E18.5 TC mutant at the level of the red nucleus double-stained for CC3 (magenta) and GAD67 (green). Massive apoptosis occurred at the reticular formation.
(B-E) In situ hybridization (ISH) with α12 (B), β22 (C), γA3 (D), and γC4 (E) antisense probes in the coronal sections of the midbrain of a control mouse (+/+:TGtaf7). Sections in B-E were all neighboring sections from the same brain.
(F-T) Magnified view of the three representative brain areas showing massive apoptosis: septum (F, I, L, O, R), cortical amygdala (G, J, M, P, S), and reticular formation in the midbrain (H, K, N, Q, T). (F-H) Double immunostaining for CC3 (magenta) and GAD67 (green). Rectangles are approximate visual fields of images in (I-T). (I-T) ISH of γC4 (I-K), α12 (L-N), β22 (O-Q), and γA3 (R-T) antisense probes in the coronal sections of E18.5 control brain corresponding to the rectangular area in (F-H). ISH signal clearly showed the dominant expression of γC4 in the vast majority of cells and the sparse expression of α12, β22, and γA3 stochastic isoforms. Scale bars: 200 μm in (A), 200 μm in (B-E), 100 μm in (F-H), and 100 μm in (I-T).