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Fig. 3. XKCM1 and NuMA are localized to the MTOC-TMA and meiotic spindles during maturation. Maturing oocytes were fixed and stained with
anti- -tubulin (A, D), anti-XKCM1 (B, C, E, G, I), anti-NuMA (J–L), and TO-PRO-3 to visualize chromosomes (F, H). (A) At WSF, MTs of the
MTOC-TMA were observed extending from the transient MTOC near the vegetal surface of the GV into the nucleoplasm. (B) Anti-XKCM1 stained the base
of the MTOC-TMA and faintly stained MTs of the TMA extending into the nucleoplasm (a projection of 11 optical sections). (C) XKCM1 antibodies stained
the base of a late MTOC-TMA as it approached the animal cortex (0–15 min after WSF, a single optical section). (D–F) XKCM1 colocalized with MTs of
the compacted meiotic spindle (15–30 min after WSF, a projection of 14 optical sections). XKCM1 antibodies also stained foci associated with condensed
meiotic chromosomes that may correspond to the centromeres (arrows). (G–I) XKCM1 is localized to centromeres and MTs of the first meiotic spindle at
30–45 min after WSF (arrows, projection of 17 optical sections). (J) Anti-NuMA stains the base of the MTOC-TMA at 0–20 min after WSF in this single
optical section. (K) NuMA is localized to the compacting meiotic spindle 15–30 min after WSF (a projection of 10 optical sections). (L) Antibodies against
NuMA stain the poles of the first meiotic spindle 30–45 min after WSF (a projection of 7 optical sections). Scale bars are 50 m (A, B, and J), 10 m (C–H,
K, and L), and 2 m (I). |
