Figure 3

Tracking GFP CETN2 centriole loss from PGCs through maturation to metaphase-II arrest. Nested bar depiction of oocyte populations in PGCs, oogonia, adolescent (P4) primary oocytes, various adult oocytes during follicular growth, and meiotic oocytes showing the percentage of GFP CETN2 foci as doublets (1 pair), two doublets (2 pair), or > two doublets (i.e. more than 3 doublets) at each stage. Tightly apposed GFP CETN2 doublets in association with PCM are observed in all e11.5 PGCs, fetal stage oogonia (e14.5 and e18.5), and neonate early oocytes prior to sexual maturity (graph: a-c, red bars; image panel: 3A–C, green). In adult ovaries, however, follicular recruitment under the influence of sex hormones alters the pattern of GFP CETN2 doublets during oocyte growth to full maturity, reducing both the number of doublets (graph: d-e, red bars) and their tight association (image panel: D–E, green; arrows point to GFP CETN2 doublets; arrowheads: GFP aggregates). GFP CETN2 doublets continue to decline with the onset of meiotic maturation (graph: f-h, red bars), with a significant increase in GFP CETN2 doublets or single pairs within the PCM at a single spindle pole in metaphase-I and -II oocytes (graph: f-h, blue bars; image panel: F–H, green). Other GFP CETN2 configurations include multiple doublets observed in immature, fully grown, and early prometaphase-I oocytes (graph: d-f, green bars), but these are significantly decreased by the end of first meiosis and not visible in metaphase-II-arrested oocytes. Minimum of three trials, except for e14.5 and primary follicles (two trials). Image panel: direct detection of GFP CETN2 doublets and singles after fixation and co-labeling with PCM antibody (γ-tubulin or pericentrin; not shown) and DNA (not shown). Images are at similar exposure and size for comparative analysis. The drop in GFP signal background (F–H) reflects dilution of GFP signal as the oocyte volume increases during follicular growth. Bar = 0.4 µm.