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Fig. 6. Alk2 is essential for MIS signaling, transition of
Misr2 expression and Müllerian duct regression in the rat.
E14.5 male urogenital ridges were treated with control-siRNA (A,C,E) or
Alk2-siRNA (B,D,F) for 10 hours. (A,B) Cultured for
additional 10 hours followed by whole-mount immunofluorescence analysis of
activated R-SMAD1, 5, 8 (P-SMAD). (C,D) Cultured for an
additional 20 hours followed by in situ hybridization to detect
Misr2. (E-H) Cultured for an additional 48 hours followed by
in situ hybridization to detect Wnt7a expression. White arrows
indicate the cranial regions with high (A) or low (B) P-SMAD expression for
comparison. The presence (C) or absence (D) of Misr2 expression can
be noted in the regions between the Müllerian and Wolffian ducts
(arrowheads). Black arrows indicate the persistence of Wnt7a
expression in the remaining Müllerian duct epithelium (F,H). The position
of transverse sections (G,H) is marked by broken lines on E and F,
respectively. Cranial is oriented towards the top and Müllerian duct to
the right of individual images. M, Müllerian duct; T, testis; W, Wolffian
duct. Scale bar: 500 µm for all the whole-mount samples; 50 µm for all
the sections.
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