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Fig. 1. A: DMg immunoblot of Xenopus brain. In brain homogenates from stage 55/56, DMg antibody detects a single band of 35 kDa and a faint lower molecular band. This lower molecular weight band as well as a very weak band in stage 63 were not absorbed by DMg–glutathione-S-transferase fusion proteins, revealing that these bands were nonspecific. In stage 42 forebrain (B,C) and hindbrain (D,E), DMg immunoreactivity is detected in radially oriented cells, extending from ventral surface to pial surface. At higher magnification (C, shown in B by box), cells are observed, which have their cell bodies facing the ventricle and which extend their processes toward the pial surface. At the pial surface of the hindbrain (E, shown in D by box), the endfeet and connected processes are positive for DMg. F–K: Double immunolabelling of DMg and vimentin in stage 53 hindbrain. DMg and vimentin are coexpressed in some laterally extending processes (arrowheads). On the other hand, the processes extending both at the floor and roof plates, in which vimentin is strongly positive, do not show DMg immunoreactivity (arrows). Scale bars 5 50 mm in B,D; 20 mm in C,E; 100 mm in H (applies to F–H); 50 mm in K (applies to I–K). |
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Fig. 4. DMg, vimentin, and glial fibrillary acidic protein (GFAP) localization on stage 45/46 hindbrain. A: Hoechst nuclear staining. Arrowheads indicate the rhombomere centers. B: DMg-positive radial glial processes are observed throughout the hindbrain. Segmental distribution of radial glia, if any, is not evident. C: Vimentin-positive cells start arranging in a segmental manner. D: GFAP-positive cells are extensively colocalized with vimentin, except the longitudinal fibers are strongly positive for GFAP (arrow). Scale bar 5 20 mm in A (applies to A–D). |
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Fig. 5. DMg mRNA, DMg, vimentin, and glial fibrillary acidic protein (GFAP) protein localization in stage 53/54 hindbrain longitudinal sections. A: DMg transcript is concentrated in the center of rhombomere (arrowheads). B: In contrast, DMg-positive radial glial processes distribute in the boundaries of the rhombomere and are occasionally seen in the center region. C: Vimentin-positive radial glial processes distribute in a similar manner as that of DMg. D: However, GFAP immunoreactivity is strongly concentrated in the center of each rhombomere and distribute throughout the white matter. Scale bars 5 20 mm in A–D. |
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Fig. 6. A: DiI is injected into the ventricle of the stage 53/54
hindbrain. DiI uptake is exclusively seen in the radial glia at the
centers (A). B: These processes are almost exclusively derived from
the region where DMg-positive processes are found. The processes of
the boundary radial glia extend upward in a fan-shaped manner (B).
C,D: Vimentin and glial fibrillary acidic protein (GFAP) localization
in stage 60 hindbrain. Vimentin is mainly detected at the borders (C),
whereas GFAP is localized in the center (D). Arrowheads indicate
center regions. Scale bars 5 20 mm in B (also applies to A) and D (also
applies to C). |
