Figure 1.
Leader cell nuclei transiently deform during the onset of border cell migration. (A) Overview of the Drosophila ovary, ovariole, and egg chamber stages. (B) Illustration of the border cells (green) and polar cells (yellow). (C) A max projection of images from a time lapse of a stage 9 egg chamber to show the border cell cluster movement over time. The egg chamber is shown with differential interference contrast and border cells expressing GFP-moesin are labeled with a different color for each time point. (D) A stage 9 egg chamber expressing fruitlessGal4;UAS-GFP-moesin (border cells);UASdsRED.NLS (nuclei) and incubated with 10 kDa dextran-Alexa647 to label junctures between cells. (D′and D″) The inverted fluorescent image of the 10 kDa dextrans in both XY and YZ views to show possible paths for border cells to migrate through. Arrowheads mark central extracellular spaces where the border cells migrate. Scale bars: 10 μm. (E) Individual and mean ± SEM values for diameters of the indicated features. n = 10 (clusters), 58 (cells and nuclei), 63 (>3 nurse cell [NC] junctures), 10 (initial juncture). One-way ANOVA followed by a post-hoc Tukey was performed, all diameters are significantly different from one another, P < 0.0001 for all comparisons except initial juncture versus >3 NC juncture, P = 0.004. Measurements are from N = 10 egg chambers. (F and G) XY (F) and YZ (G) images of border cells and junctures from a time-lapse series labeled with the indicated markers. Yellow box marks the same juncture shown in F and G. White box in F shows the leading cell’s nucleus in grayscale. (H) Plot showing individual juncture widths relative to time of protrusion entry (green) and nuclear entry (magenta) normalized to the size of the initial juncture. A mixed effects model (REML) followed by Tukey’s multiple comparison test was performed for statistics. N = 10 egg chambers. (I and J) Example images from time-lapse series of border cell nuclei (I) or polar cell nuclei (J) labeled with UAS-dsRED.NLS expressed by c306Gal4 (I) or UpdGaL4 (J). Magenta arrowheads label the lead cell’s nucleus. (K and L) Plots showing individual and average ± SEM nuclear aspect ratios (K) and changes in nuclear aspect ratio (L) for each individual nucleus. n = 20 (leader), 43 (follower), and 24 (polar) cell nuclei. One-way ANOVA followed by Tukey post-hoc was performed, data were collected from N = 10 movies for border cells and N = 12 movies for polar cells. Scale bars: 10 μm. Genotypes and experimental replicates are reported in Table S2.

Leader cell nuclei transiently deform during the onset of border cell migration. (A) Overview of the Drosophila ovary, ovariole, and egg chamber stages. (B) Illustration of the border cells (green) and polar cells (yellow). (C) A max projection of images from a time lapse of a stage 9 egg chamber to show the border cell cluster movement over time. The egg chamber is shown with differential interference contrast and border cells expressing GFP-moesin are labeled with a different color for each time point. (D) A stage 9 egg chamber expressing fruitlessGal4;UAS-GFP-moesin (border cells);UASdsRED.NLS (nuclei) and incubated with 10 kDa dextran-Alexa647 to label junctures between cells. (D′and D″) The inverted fluorescent image of the 10 kDa dextrans in both XY and YZ views to show possible paths for border cells to migrate through. Arrowheads mark central extracellular spaces where the border cells migrate. Scale bars: 10 μm. (E) Individual and mean ± SEM values for diameters of the indicated features. n = 10 (clusters), 58 (cells and nuclei), 63 (>3 nurse cell [NC] junctures), 10 (initial juncture). One-way ANOVA followed by a post-hoc Tukey was performed, all diameters are significantly different from one another, P < 0.0001 for all comparisons except initial juncture versus >3 NC juncture, P = 0.004. Measurements are from N = 10 egg chambers. (F and G) XY (F) and YZ (G) images of border cells and junctures from a time-lapse series labeled with the indicated markers. Yellow box marks the same juncture shown in F and G. White box in F shows the leading cell’s nucleus in grayscale. (H) Plot showing individual juncture widths relative to time of protrusion entry (green) and nuclear entry (magenta) normalized to the size of the initial juncture. A mixed effects model (REML) followed by Tukey’s multiple comparison test was performed for statistics. N = 10 egg chambers. (I and J) Example images from time-lapse series of border cell nuclei (I) or polar cell nuclei (J) labeled with UAS-dsRED.NLS expressed by c306Gal4 (I) or UpdGaL4 (J). Magenta arrowheads label the lead cell’s nucleus. (K and L) Plots showing individual and average ± SEM nuclear aspect ratios (K) and changes in nuclear aspect ratio (L) for each individual nucleus. n = 20 (leader), 43 (follower), and 24 (polar) cell nuclei. One-way ANOVA followed by Tukey post-hoc was performed, data were collected from N = 10 movies for border cells and N = 12 movies for polar cells. Scale bars: 10 μm. Genotypes and experimental replicates are reported in Table S2.

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