Abstract
In the developing CNS, asymmetric cell division is critical for maintaining the balanced production of differentiating neurons while renewing the population of neural progenitors. In invertebrates, this process depends on asymmetric inheritance of fate determinants during progenitor divisions. A similar mechanism is widely believed to underlie asymmetrically fated divisions in vertebrates, but compelling evidence for this is missing. We used live imaging of individual progenitors in the intact zebrafish embryo CNS to test this hypothesis. We found that asymmetric inheritance of a subcellular domain is strongly correlated with asymmetric daughter fates and our results reveal an unexpected feature of this process. The daughter cell destined to become a neuron was derived from the more apical of the two daughters, whereas the more basal daughter inherited the basal process and replenished the apical progenitor pool.
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Acknowledgements
We thank C. Araya, C. Buckley, K. Cerveny, G. Girdler, A. Lumsden, X. Ren, M. Tada, M. Tawk and M. Wassef for discussions and comments on the manuscript and the University College London and King's College London fish facilities for providing and taking care of the fish. We thank F. Mahmood for the aPKC immunostaining. P.A. was supported by an European Molecular Biology Organization fellowship (ALTF-764-2005) and an EU Marie Curie fellowship (IEF 039695).
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P.A. and J.D.W.C. conceived the project, planned the experiments and wrote the manuscript. P.A. generated and analyzed most of the data and produced the Par3-RFP construct. A.M.R. made the Par3-GFP and Numb-GFP constructs and generated data on Numb. D.B. analyzed the size distribution of the apical end feet (Supplementary Fig. 1). E.B. carried out the statistical analysis.
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Supplementary Text and Figures
Supplementary Figures 1–8 (PDF 13638 kb)
Supplementary Movie 1
Time-lapse of two dividing cells labeled with membrane-GFP followed through to their fates; the cell on the left produces daughters with asymmetric fates (NP) and the cell on the right has daughters with symmetric fates (PP). White asterisk indicates neuronal detachment from the apical surface. (MOV 1044 kb)
Supplementary Movie 2
Symmetric inheritance of Par3-GFP during cell division. (MOV 108 kb)
Supplementary Movie 3
Asymmetric inheritance of Par3-GFP during cell division. (MOV 44 kb)
Supplementary Movie 4
Three-dimensional reconstruction and rotation of the cell shown in Movie 3 to illustrate asymmetric inheritance of Par3-GFP. (MOV 779 kb)
Supplementary Movie 5
Numb-GFP localization through out division. Numb-GFP localizes at the basal lateral cortex of dividing progenitors and becomes punctate after cell division. (MOV 193 kb)
Supplementary Movie 6
Three-dimensional reconstruction and rotation of a pair of daughters immediately after mitosis illustrating asymmetry of Numb-GFP inheritance. One daughter (initially on left hand side) inherits Numb-GFP all around its cortex while the other has a distinct Numb-GFP–free zone that corresponds to the progenitors apical domain as marked by Par3-RFP. (MOV 1624 kb)
Supplementary Movie 7
Asymmetric inheritance of Par3-GFP producing asymmetrically fated daughters (NP). The daughter cell that inherits Par3-GFP domain (white arrow) becomes a neuron (white arrow at the end of the time lapse). The non-apical daughter (blue asterisk) divides at the end of the time lapse (blue arrows indicate the two daughter cells). (MOV 2047 kb)
Supplementary Movie 8
Three-dimensional reconstruction and rotation of the cell shown in Movie 7 to illustrate asymmetric inheritance of Par3- GFP. (MOV 789 kb)
Supplementary Movie 9
Asymmetric inheritance of Par3-GFP with near perpendicular cleavage producing asymmetrically fated daughters (NP). The neuron derives from the daughter that inherits Par3-GFP. The daughter cell that inherits Par3-GFP domain (white arrow) becomes a neuron (white arrow at the end of the time lapse). The non-apical daughter (blue asterisk) divides (blue arrow) at the end of the time lapse. (MOV 795 kb)
Supplementary Movie 10
Three-dimensional reconstruction and rotation of the cell shown in Movie 9 to illustrate asymmetric inheritance of Par3-GFP. (MOV 1320 kb)
Supplementary Movie 11
Three-dimensional reconstruction and rotation of a dividing cell labelled with membrane-GFP to illustrate non-apical daughter cell (left hand side) is initially detached from apical surface but then forms a new apical attachment (dotted line labels apical). (MOV 1631 kb)
Supplementary Movie 12
Three-dimensional reconstruction of two time points from Par3-RFP labelled division (see also Fig. 6b). The two cells on the left of this cluster were generated by a division 6 minutes previously. In the first time point the left hand daughter does not have a Par3-RFP domain but this daughter generates an apical Par3-RFP domain by the second time point 9 minutes later. The third cell on the right of the cluster did not divide in the period of the time lapse (yellow asterisk). (MOV 2206 kb)
Supplementary Movie 13
Progenitor with split basal process. The basal process retracts from one daughter so that a single basal process is inherited by only one daughter cell. (MOV 169 kb)
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Alexandre, P., Reugels, A., Barker, D. et al. Neurons derive from the more apical daughter in asymmetric divisions in the zebrafish neural tube. Nat Neurosci 13, 673–679 (2010). https://doi.org/10.1038/nn.2547
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DOI: https://doi.org/10.1038/nn.2547
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