Fig. 3: Chromosome alignment is reduced and delayed in Cenp-V−/− oocytes. | Nature Communications

Fig. 3: Chromosome alignment is reduced and delayed in Cenp-V−/− oocytes.

From: CENP-V is required for proper chromosome segregation through interaction with spindle microtubules in mouse oocytes

Fig. 3

Polar body extrusion and chromosome alignment return to control values upon Cenp-V-Gfp mRNA injection. (a) Selected time frames from a time-lapse experiment showing progression through meiotic prophase until metaphase II in Cenp-V+/+ and Cenp-V−/− oocytes expressing β-TUBULIN-GFP (magenta), H2B-RFP (cyan, large image in grey) and/or CENP-V-GFP (green) after mRNA injection. At 09:30 h in metaphase I, the spindle is assembled and chromosomes are aligned in the equatorial plate (white dash line) in Cenp-V+/+ compare to Cenp-V−/− oocyte. Note that Cenp-V−/− oocytes show normal chromosome alignment and extrude the PB only when expressing CENP-V-GFP. Oocytes were imaged after 4 h release from GV every 30 m. Maximum intensity projection is shown. See also supplementary Movies S2,S3,S4). Scale bar = 10 µm. b Percentage of cells that show chromosome alignment overtime of the experiment shown in (a). in Cenp-V+/+ and Cenp-V−/− oocytes. Quantification finishes before the first polar body extrusion. Note that at 10 h, 75 % of control oocytes have reached full chromosome alignment compared to the 35% of Cenp-V−/− oocytes, and only the injection of Cenp-V-Gfp (green square line) mRNA restores the values to the control. c Cumulative percentage of first polar body extrusion overtime of the experiment shown in (a). Note that Cenp-V−/− oocytes properly extrude the PB only when expressing CENP-V-GFP (green square line) and not when expressing β-TUBULIN-GFP/H2B-RFP. See also supplementary movie S1. Statistic: (b, c) data were expressed as mean ± SD and statistical differences were tested by a two way Anova test (***p < 0.001, **p < 0.01, *p < 0.05). The exact p values are available in the source data file. Statistical differences refer to the average of all data points. Anova test between Cenp-V+/+ and Cenp-V+/+ injected was not significant. Anova test between Cenp-V−/− and Cenp-V−/− β-Tubulin-Gfp/H2B-Rfp injected was not significant. n = 249 cells from 5 different experiments.

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