Fig. 1: CENP-V localises to the centromere regions during meiotic prophase and to MTOCs, chromosomes, and spindle during oocyte maturation.
a CENP-V distribution during meiotic prophase in Cenp-V+/+ embryonic chromosome spreads by immunofluorescent staining. Chromosome axes (AEs/LEs) were stained with anti-SYCP3 (blue); CENP-V was probed with anti-CENP-V (green); centromeres and surrounded regions by anti-ACA. CENP-V signal started to appear at pachytene and concentrates at the centromere regions (yellow arrows) at diakinesis; n = 19 cells from 2 independent experiments. Scale bar = 5 μm. b CENP-V distribution during mouse oocyte maturation in whole cells. Live imaging performed with Cenp-V+/+ oocytes injected by mRNA of H2B-Rfp (blue) and Cenp-V-Gfp (green). Maximum projections from representative oocytes at germinal vesicle break down (GVBD), metaphase I, and metaphase II stages are shown. Scale bar = 10 μm. Some GVBD cells were fixed and immunostained with anti γ-Tubulin as an MTOC marker. Spindle at metaphase I and II were visualised by SiR Tubulin dye. Polar body is indicated as PB. Note that at the early stages of maturation CENP-V distributes along the entire chromosomes and colocalizes with MTOCs (yellow arrow heads). At metaphase I and II CENP-V appears mainly in the spindle and enriches at the spindle poles. See also supplementary movie 1; n = 12 cells from 3 independent experiments. c Quantification of colocalization of CENP-V with the spindle pole by determining the respective Pearson’s correlation coefficient; n = 10 cells from 3 independent experiments; the color code highlights the frequency of dots present in a certain region of the scatter plot (from blue to yellow and white with increasing frequencies).
