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Rotation and asymmetry of the mitotic spindle direct asymmetric cell division in the developing central nervous system

Nature Cell Biology volume 2, pages 7–12 (2000)Cite this article

Abstract

The asymmetric segregation of cell-fate determinants and the generation of daughter cells of different sizes rely on the correct orientation and position of the mitotic spindle. In the Drosophila embryo, the determinant Prospero is localized basally and is segregated equally to daughters of similar cell size during epidermal cell division. In contrast, during neuroblast division Prospero is segregated asymmetrically to the smaller daughter cell. This simple switch between symmetric and asymmetric segregation is achieved by changing the orientation of cell division: neural cells divide in a plane perpendicular to that of epidermoblast division. Here, by labelling mitotic spindles in living Drosophila embryos, we show that neuroblast spindles are initially formed in the same axis as epidermal cells, but rotate before cell division. We find that daughter cells of different sizes arise because the spindle itself becomes asymmetric at anaphase: apical microtubules elongate, basal microtubules shorten, and the midbody moves basally until it is positioned asymmetrically between the two spindle poles. This observation contradicts the widely held hypothesis that the cleavage furrow is always placed midway between the two centrosomes.

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Figure 1: Drosophila neuroblasts divide in an axis perpendicular to that of epidermoblast division.
Figure 2: The neuroblast spindle rotates by 90° at metaphase.
Figure 3: Loss of Inscuteable impairs spindle rotation.
Figure 4: The mitotic spindle becomes asymmetric during neuroblast cell division.
Figure 5: Neuroblast centrosomes become asymmetric at anaphase.
Figure 6: Spindle dynamics during neuroblast and epidermoblast cell division.

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Acknowledgements

We thank J. Raff and C. Waterman-Storer for discussions; J. Haseloff, J. Raff, H. Chang and G. Rubin for providing DNA constructs, antibodies and/or Drosophila lines; T. Bossing, J. Gray, J. Raff and P. van Roessel for comments on the manuscript; and A. Sossick for technical assistance in video production. J.A.K. is a Wellcome Trust Prize Student. This work was funded by the Wellcome Trust.

Correspondence and requests for materials should be addressed to A.H.B.

Supplementary information is available on Nature Cell Biology’s World-Wide Web site (http://cellbio.nature.com).

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Authors and Affiliations

  1. Wellcome/CRC Institute and Department of Genetics University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK

    Julia A. Kaltschmidt, Catherine M. Davidson & Andrea H. Brand

  2. Wellcome/CRC Institute and Department of Anatomy, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK

    Nicholas H. Brown

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Correspondence to Andrea H. Brand.

Supplementary information

Movie 1 Cell division in epidermoblasts.

In epidermoblasts, the centrosome duplicates on the basal side of the nucleus, and the two centrosomes migrate laterally to opposite sides of the cell. The spindle forms parallel to the surface of the embryo and remains in this orientation throughout mitosis. Microtubules were labelled with tau-GFP, and confocal images were recorded every 12 s. Basal is at the top and apical at the bottom. Lateral view of a stage-10 embryo (also see Fig. 2 of article). (MOV 2702 kb)

Movie 2 Apical centrosome duplication at interphase.

Although the spindle can rotate in either direction, the direction of rotation usually correlates with the position of the centrosome at interphase. When the interphase centrosome duplicates apically, the primed centrosome moves to the more anterior side of the cell before moving apically. The mitotic spindle rotates 90° to a position along the apical-basal axis of the cell. Microtubules were labelled with tau-GFP, and confocal images were recorded every 12 s. Basal is at the top and apical at the bottom. Lateral view of a stage-10 embryo. (MOV 3811 kb)

Movie 3 Basal centrosome duplication at interphase.

Although the spindle can rotate in either direction, the direction of rotation usually correlates with the position of the centrosome at interphase. When the interphase centrosome duplicates basally, the primed centrosome first migrates to the more posterior side of the cell at prophase, and then apically at metaphase. The mitotic spindle rotates 90° to a position along the apical-basal axis of the cell. Microtubules were labelled with tau-GFP and cell cortices with Src-GFP. Images were collected every 20 s. Basal is at the top and apical at the bottom. Lateral view of a stage-10 embryo. (MOV 2490 kb)

Movie 4 The neuroblast spindle rotates 90° at metaphase.

In most cases the spindle is formed before rotation. In this case the bipolar spindle forms as it rotates 90° to its position along the apical-basal axis of the cell. Microtubules were labelled with tau-GFP and cell cortices with Src-GFP. Images were taken every 4 s. Basal is at the top and apical at the bottom. Lateral view of a stage-11 embryo (also see Fig. 2 of article). (MOV 1944 kb)

Movie 5 Loss of Inscuteable impairs spindle rotation.

In embryos injected with inscuteable dsRNA, spindle rotation fails to occur in 10% of neuroblasts. At first the spindle forms normally, perpendicular to the apical-basal axis. The spindle seesaws but does not rotate. In this example, cell division occurs at a 45° angle to the apical-basal axis. Microtubules were labelled with tau-GFP and cell cortices with Src-GFP. Images were taken every 10 s. Basal is at the top and apical at the bottom. Lateral view of a stage-10 embryo (also see Fig. 3 of article). (MOV 2650 kb)

Movie 6

The mitotic spindle becomes asymmetric during neuroblast cell division. The neuroblast mitotic spindle is symmetric and centrally placed at metaphase. At the start of anaphase, the basal cell membrane starts to invaginate and the spindle becomes asymmetric. The distance between the apical and basal centrosome stays the same throughout the division (white arrows mark their starting position and white dots their end point). The midbody, however, gradually moves basally (red arrow), until it is positioned at the site of the cleavage furrow. The neuroblast aster enlarges from anaphase onwards, and the GMC aster is reduced, while the neuroblast half of the spindle elongates and the GMC half becomes shorter. Microtubules were labelled with tau-GFP and cell cortices with Src-GFP. Confocal images were taken every 5 s. Basal is at the top and apical at the bottom. Lateral view of a stage-10 embryo (also see Fig. 4 of article). (MOV 1366 kb)

Movie 7 The mitotic spindle seesaws in epidermoblasts.

The mitotic spindle seesaws in epidermoblasts before undergoing cell division. Microtubules were labelled with tau-GFP, and confocal images were recorded every 20 s. Ventral view of a stage-11 embryo. Anterior is to the left, posterior to the right. (MOV 3857 kb)

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Kaltschmidt, J., Davidson, C., Brown, N. et al. Rotation and asymmetry of the mitotic spindle direct asymmetric cell division in the developing central nervous system. Nat Cell Biol 2, 7–12 (2000). https://doi.org/10.1038/71323

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