![Nuclear crowding in a common epithelial plane after MTdisruption. (A) 3D reconstruction of cell membranes (pink) and nuclei (blue) from live imaging data of control (vehicle-injected, top panel) or nuclear dispersion defective (colchicine-injected [MT-], bottom panel) embryos. Slanted view showing highly featured nuclear surface (left), side view (mid), and top view (right) showing nuclei flattened for optimal packing in MT- embryos (related to Video 7). (B) Probability of nuclear position relative to neighboring nuclei at pre- (−5 to 0 min, blue), early (0–10 min, yellow), and mid- (15–20 min, red) GBE; n = 10,781 pairs of nuclei from k = 3 embryos. (C) Orthogonal views showing decreased nuclear crowding in control (compare 0 min to 10 and 20 min) and enhanced nuclear crowding in MT- embryos (compare 0 min to 10 and 20 min). (D) Still image showing pavement stone-like phenotype in dispersion defective MT- embryos at late GBE (20 min) (compare to control) (related to Video 6). Nuclei are color-coded according to their major to minor axis ratio. Image slice from ∼12 µm below the apical surface. (E) The ratio of nucleus area to cell area at the nuclear midplane in MT- embryos compared at early (0 min) and mid- (+20 min) GBE; n = 290 cells and nuclei from k = 3 embryos. (F) Peak nuclear velocities in control and MT- embryos; n = 769 nuclei for control from k = 4 embryos and n = 325 nuclei for MT- from k = 3 embryos for each background. (G) Maximum packing ratio achieved by clusters of nuclei as measured at pre- (−5 min) and mid- (+20 min) GBE for MT- embryos; n = 420 clusters of cells from k = 3 embryos. (H) Average cytoplasmic area ratio available in clusters of cells at the midplane of a given central cell’s nucleus in control (red) and MT- (blue) embryos during GBE (left). Available cytoplasmic area at −5 min (pre) and +20 min (mid) after the onset of GBE (right); n = 370 control cell clusters from k = 4 embryos and n = 120 clusters for MT- from k = 3 embryos for each background. Error envelopes indicate SD. The measured n values are regardless of time points where not indicated. Scale bar = 10 µm for C and D. Statistical significance was calculated with Student’s t test in E and Mann–Whitney U-test in F–H. ns = not significant, ****P < 0.0001.](https://cdn.rupress.org/rup/content_public/journal/jcb/223/12/10.1083_jcb.202405078/2/jcb_202405078_fig6.png?Expires=1767803070&Signature=dYXsykoq2sFFTio8Qm7rUEDdifnnz66RLaxEwjIEvctCqwEWuY0A09qsYcdpgnKz6iAsOAsmegOE-LDM2vzUc475nxPL8Jc2BorlI-DCToY23QI0l5PGJLj7sUDSXdiyoPjdTJzN9nZ6FJ0LrduEuhST4pLn9C5YqKFlqGdfM~5MQo~fwCBb~F8EXMcAnp-nCYqnLXHK7RqUL-slMHtzr~rf5JJKBPvYimc~bqxZyPMsLgUvgz21KTwCGroa3KW11-dg-A50297qTaEr4wgSRK74lV4sazjf6xtQZ-rXhH7QTTNc8LiVR0Q7BxgROPiKOoLqrHwE0K5i77DxM4kVVA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Nuclear crowding in a common epithelial plane after MT disruption. (A) 3D reconstruction of cell membranes (pink) and nuclei (blue) from live imaging data of control (vehicle-injected, top panel) or nuclear dispersion defective (colchicine-injected [MT-], bottom panel) embryos. Slanted view showing highly featured nuclear surface (left), side view (mid), and top view (right) showing nuclei flattened for optimal packing in MT- embryos (related to Video 7). (B) Probability of nuclear position relative to neighboring nuclei at pre- (−5 to 0 min, blue), early (0–10 min, yellow), and mid- (15–20 min, red) GBE; n = 10,781 pairs of nuclei from k = 3 embryos. (C) Orthogonal views showing decreased nuclear crowding in control (compare 0 min to 10 and 20 min) and enhanced nuclear crowding in MT- embryos (compare 0 min to 10 and 20 min). (D) Still image showing pavement stone-like phenotype in dispersion defective MT- embryos at late GBE (20 min) (compare to control) (related to Video 6). Nuclei are color-coded according to their major to minor axis ratio. Image slice from ∼12 µm below the apical surface. (E) The ratio of nucleus area to cell area at the nuclear midplane in MT- embryos compared at early (0 min) and mid- (+20 min) GBE; n = 290 cells and nuclei from k = 3 embryos. (F) Peak nuclear velocities in control and MT- embryos; n = 769 nuclei for control from k = 4 embryos and n = 325 nuclei for MT- from k = 3 embryos for each background. (G) Maximum packing ratio achieved by clusters of nuclei as measured at pre- (−5 min) and mid- (+20 min) GBE for MT- embryos; n = 420 clusters of cells from k = 3 embryos. (H) Average cytoplasmic area ratio available in clusters of cells at the midplane of a given central cell’s nucleus in control (red) and MT- (blue) embryos during GBE (left). Available cytoplasmic area at −5 min (pre) and +20 min (mid) after the onset of GBE (right); n = 370 control cell clusters from k = 4 embryos and n = 120 clusters for MT- from k = 3 embryos for each background. Error envelopes indicate SD. The measured n values are regardless of time points where not indicated. Scale bar = 10 µm for C and D. Statistical significance was calculated with Student’s t test in E and Mann–Whitney U-test in F–H. ns = not significant, ****P < 0.0001.