FIGURE 1.

Identification of pinwheel structures in GFAP-TK mouse spinal cord and SVZ-derived neurospheres cultured in vitro (A) In spinal cord-derived (SC) neurospheres cultured in vitro, an acetylated tubulin (green) antibody was used to mark cilia, a GFAP (blue) antibody was used to mark astrocytes, a CD24 (red) antibody was used to mark ependymal cells. Central clusters of acetylated tubulin and GFAP detected are indicated by a continuous line, and the periphery of CD24-positive ependymal cells and the pinwheel structure is labeled by dashed lines (schematic). Lateral extensions of astrocytes are indicated by white arrows (schematic). (B) In spinal cord-derived (SC) neurospheres cultured in vitro, a β-catenin (green) antibody delineates the cell membrane, and an antibody against γ-tubulin (red) was used to detect single basal bodies marked by an empty arrowhead in small cells (as delineated by β-catenin). The γ-tubulin antibody also detected groups of basal bodies (marked by arrows) or double basal bodies (marked by arrowheads) in large cells (as delineated by β-catenin). Central clusters of γ-tubulin-positive small cells are indicated by a continuous line, and the periphery of the pinwheel structure is labeled by dashed lines (schematic). Single ependymal cells that are shared by two adjacent pinwheels are labeled by double-head arrows (i). (C) In SVZ-derived neurospheres cultured in vitro, an acetylated tubulin (green) antibody was used to mark cilia, a GFAP (blue) antibody was used to mark astrocytes, a CD24 (red) antibody was used to mark ependymal cells and DAPI was used to mark nuclei (gray). Central clusters of acetylated tubulin and GFAP detected are indicated by a continuous line, and the periphery of CD24-positive ependymal cells and the pinwheel structure is labeled by dashed lines (schematic). Lateral extensions of astrocytes are indicated by white arrows (schematic). The scale bars correspond to 20 (A,B) or 10 μm (C).