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Figure 4 | BMC Neuroscience

Figure 4

From: Shox2 is required for the proper development of the facial motor nucleus and the establishment of the facial nerves

Figure 4

Nestin-Cre driven conditional Shox2-deletion results in disruptions in the facial motor nucleus and changes in cell death. a, b Ventral view of the P0 brain of control (a) and Nestin-Cre; Shox2 flox/− (b) animals stained with the 2H3 antibody demonstrate that the facial motor nucleus (nVII) is severely reduced in Nestin-Cre; Shox2 flox/− pups (compare a to b, dashed-circle and arrows). cf X-gal staining of P0 control (c, e) and Nestin-Cre; Shox2 lacZ/flox (d, f) pup brains (viewed ventrally) show Shox2 expression in the facial motor nucleus and demonstrate that the nucleus is severely reduced in size (compare e to f, arrows) in mice lacking Shox2 function. Cleaved active-CASP3 immunostaining on E12.5 (g, h), E14.5 (j, k), E16.5 (m, n) and P0 (p, q) control (g, j, m, p) and Nestin-Cre; Shox2 flox/ mutant (h, k, n, q) sagittal sections through the facial motor nucleus (nVII). i Quantification of the number of cleaved active-CASP3 + cells present in the E12.5 facial motor nucleus (see g, h) of control and Nestin-Cre; Shox2 flox/ embryos (plotted values are the mean ± S.E.M, 3 sections per animals, n = 3 animals, p = 0.0038). l Quantification of the number of cleaved active-CASP3 + cells present in the E14.5 facial motor nucleus (see j, k) of control and Nestin-Cre; Shox2 flox/ embryos (plotted values are the mean ± S.E.M, 3 sections per animals, n = 3 animals, p = 0.0007). o Quantification of the number of cleaved active-CASP3 + cells present in the E16.5 facial motor nucleus (see m, n) of control and Nestin-Cre; Shox2 flox/ embryos (plotted values are the mean ± S.E.M, 3 sections per animals, n = 3 animals, p = 0.1929). r Quantification of the number of cleaved active-CASP3 + cells present in the P0 facial motor nucleus (see p, q, dashed-circle) of control and Nestin-Cre; Shox2 flox/ embryos (plotted values are the mean ± S.E.M, 3 sections per animals, n = 3 animals, p = 0.6875). s Quantification of the number of cleaved active-CASP3 + cells present in and around the P0 facial motor nucleus (see p, q, entire panel including cells within the dashed-circle) of control and Nestin-Cre; Shox2 flox/ embryos (plotted values are the mean ± S.E.M, 3 sections per animals, n = 3 animals, p = 0.0007). t, u Representative binary particle area diagrams from ImageJ displaying Shox2lacZ + cell staining at E14.5 (t) and P0 (u) in controls and NestinCre; Shox2 lacZ/flox facial motor nuclei. v Quantification of the area of Shox2lacZ + cells present in sagittal sections through the center of the E14.5 facial motor nucleus (see t) of control and Nestin-Cre; Shox2 lacZ/flox embryos (plotted values are the mean ± S.E.M, control = 3, Nestin-Cre; Shox2 lacZ/flox = 3, p = 0.0027). w Quantification of the area of Shox2lacZ + cells present in sagittal sections through the center of the P0 facial motor nucleus (see u) of control and Nestin-Cre; Shox2 lacZ/flox animals (plotted values are the mean ± S.E.M, control = 3, Nestin-Cre; Shox2 lacZ/flox = 3, p = 0.0011). cb cerebellum. af Scale bar 500 μm. gq Scale bar 250 μm.

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