Figure 1.
The interkinetic envelope forms between meiosis I and II in the C. elegans oocyte. (A) Representative time-lapse images of GFP::TBA-2α-tubulin (green) and mCherry::HIS-11H2B (magenta)–expressing oocytes during meiosis I and II (n = 7). Timings indicated at the bottom left corners of images are from anaphase I onset. The specific meiotic stages used for electron microscopy are highlighted in blue (mid-anaphase), orange (mid interkinesis), and purple (late interkinesis). Scale bar, 5 μm. (B) Three-dimensional reconstructions centered on chromosomes of a mid-anaphase I (left, n = 1), a mid-interkinesis (center, n = 1), and a late-interkinesis (right, n = 1) oocytes acquired by SBF-SEM. Chromosomes in magenta, membranes in contact with chromosomes in green, plasma membrane in gray, and eggshell in gold. Each reconstruction is accompanied (bottom) by a two-dimensional single section showing each chromosome set in magenta and a magnification of a region of interest (ROI) of the MII chromosomal set. Scale bar, 1 µm for the full view, 0.25 μm for the magnification. (C and D) Quantifications of the total (C) and mean (D) length of membrane contours from five reconstructed oocytes represented according to the distance between the segregating chromosomal sets (chromosome mass separation) for the PB chromosomal set (empty dots) and for the MII chromosomal set (solid dots). (E) Left: Representative time-lapse images of GFP::LEM-2LEMD2/3 and mCherry::H2B–expressing oocytes (n = 9) during meiosis I (top) and meiosis II (bottom). Timing relative to anaphase I onset is indicated at the bottom left corner of each image. The cyan arrows indicate the GFP::LEM-2LEMD2/3 plaque. Scale bar, 5 μm. Right: Quantification of the normalized GFP::LEM-2LEMD2/3 integrated intensity over time from anaphase I onset to interphase for the MII chromosomal set. Error bars correspond to the standard error of the mean. The orange and gray boxes indicate interkinesis and interphase, respectively. The same control oocyte is displayed in Fig. S2 E and Fig. S3 D. (F) Representative images centered on chromosomes of fixed oocytes showing the immunolocalization of LEM-2LEMD2/3, DNA, and Tubulin in mid anaphase I, mid and late interkinesis. Scale bar, 5 μm.

The interkinetic envelope forms between meiosis I and II in the C. elegans oocyte. (A) Representative time-lapse images of GFP::TBA-2α-tubulin (green) and mCherry::HIS-11H2B (magenta)–expressing oocytes during meiosis I and II (n = 7). Timings indicated at the bottom left corners of images are from anaphase I onset. The specific meiotic stages used for electron microscopy are highlighted in blue (mid-anaphase), orange (mid interkinesis), and purple (late interkinesis). Scale bar, 5 μm. (B) Three-dimensional reconstructions centered on chromosomes of a mid-anaphase I (left, n = 1), a mid-interkinesis (center, n = 1), and a late-interkinesis (right, n = 1) oocytes acquired by SBF-SEM. Chromosomes in magenta, membranes in contact with chromosomes in green, plasma membrane in gray, and eggshell in gold. Each reconstruction is accompanied (bottom) by a two-dimensional single section showing each chromosome set in magenta and a magnification of a region of interest (ROI) of the MII chromosomal set. Scale bar, 1 µm for the full view, 0.25 μm for the magnification. (C and D) Quantifications of the total (C) and mean (D) length of membrane contours from five reconstructed oocytes represented according to the distance between the segregating chromosomal sets (chromosome mass separation) for the PB chromosomal set (empty dots) and for the MII chromosomal set (solid dots). (E) Left: Representative time-lapse images of GFP::LEM-2LEMD2/3 and mCherry::H2B–expressing oocytes (n = 9) during meiosis I (top) and meiosis II (bottom). Timing relative to anaphase I onset is indicated at the bottom left corner of each image. The cyan arrows indicate the GFP::LEM-2LEMD2/3 plaque. Scale bar, 5 μm. Right: Quantification of the normalized GFP::LEM-2LEMD2/3 integrated intensity over time from anaphase I onset to interphase for the MII chromosomal set. Error bars correspond to the standard error of the mean. The orange and gray boxes indicate interkinesis and interphase, respectively. The same control oocyte is displayed in Fig. S2 E and Fig. S3 D. (F) Representative images centered on chromosomes of fixed oocytes showing the immunolocalization of LEM-2LEMD2/3, DNA, and Tubulin in mid anaphase I, mid and late interkinesis. Scale bar, 5 μm.

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