Molecular Cytogenetic Mapping of Chromosomal Fragments and Immunostaining of Kinetochore Proteins in Beta
Figure 1
Blue fluorescence in each panel shows the chromosomes stained with DAPI. The scale bar in (G) for the panels A-G and in (J) for the panels I and J represents 10 µm. The chromatids of the chromosome fragments are schematically contoured in panels C and F. FISH with Procumbentes-specific satellites pTS4.1 (green) and pTS5 (red ) on (A) B. procumbens; (B) PRO1; (C) the closed-up overlay of the panel (B); (D) B. patellaris; (E) PAT2; (F) the PAT2 fragment. (G) Simultaneous localization of the centromeric probes pTS5 (red) and pTS4.1 (green) on the B. procumbens meiotic chromosomes. (H) Close-up from the panel (G). (I-J) Localization of kinetochore proteins in B. vulgaris and PRO1 by immunostaining. Microtubuli are visible as green threads. Serine 10-phosphorylated histone H3 produces red signals. The right images are overlays. Microphotographs of the three-dimensional preparation were taken in different focal planes and overlaid. (I) Serine 10-phosphorylated histone H3 labels all centromeres of B. vulgaris in mitosis. The sites where the microtubuli of the spindle apparatus are attached to the centromeres are exampled by arrows. (J) PRO1 chromosomal fragment shows a H3S10- phosphorylated signal (arrowhead), thus indicating that its centromere is active in mitosis.
