Lgl, Pins and aPKC regulate neuroblast self-renewal versus differentiation


How a cell chooses to proliferate or to differentiate is an important issue in stem cell and cancer biology. Drosophila neuroblasts undergo self-renewal with every cell division, producing another neuroblast and a differentiating daughter cell, but the mechanisms controlling the self-renewal/differentiation decision are poorly understood. Here we tested whether cell polarity genes, known to regulate embryonic neuroblast asymmetric cell division1, also regulate neuroblast self-renewal. Clonal analysis in larval brains showed that pins mutant neuroblasts rapidly fail to self-renew, whereas lethal giant larvae (lgl) mutant neuroblasts generate multiple neuroblasts. Notably, lgl pins double mutant neuroblasts all divide symmetrically to self-renew, filling the brain with neuroblasts at the expense of neurons. The lgl pins neuroblasts show ectopic cortical localization of atypical protein kinase C (aPKC), and a decrease in aPKC expression reduces neuroblast numbers, suggesting that aPKC promotes neuroblast self-renewal. In support of this hypothesis, neuroblast-specific overexpression of membrane-targeted aPKC, but not a kinase-dead version, induces ectopic neuroblast self-renewal. We conclude that cortical aPKC kinase activity is a potent inducer of neuroblast self-renewal.

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Figure 1: lgl and pins regulate larval neuroblast self-renewal.
Figure 2: lgl pins double mutants have ectopic neuroblasts and fewer neurons.
Figure 3: Lgl and Pins regulate aPKC localization in larval neuroblasts.
Figure 4: Overexpression of cortical aPKC promotes neuroblast self-renewal.


  1. 1

    Betschinger, J. & Knoblich, J. A. Dare to be different: asymmetric cell division in Drosophila, C. elegans and vertebrates. Curr. Biol. 14, R674–R685 (2004)

    CAS  Article  Google Scholar 

  2. 2

    Ohlstein, B., Kai, T., Decotto, E. & Spradling, A. The stem cell niche: theme and variations. Curr. Opin. Cell Biol. 16, 693–699 (2004)

    CAS  Article  Google Scholar 

  3. 3

    Urbach, R. & Technau, G. M. Neuroblast formation and patterning during early brain development in Drosophila. BioEssays 26, 739–751 (2004)

    CAS  Article  Google Scholar 

  4. 4

    Datta, S. Control of proliferation activation in quiescent neuroblasts of the Drosophila central nervous system. Development 121, 1173–1182 (1995)

    CAS  PubMed  Google Scholar 

  5. 5

    Gateff, E. & Schneiderman, H. A. Developmental capacities of benign and malignant neoplasms of Drosophila. Roux Arch. Dev. Biol. 176, 23–65 (1974)

    CAS  Article  Google Scholar 

  6. 6

    Ohshiro, T., Yagami, T., Zhang, C. & Matsuzaki, F. Role of cortical tumour-suppressor proteins in asymmetric division of Drosophila neuroblast. Nature 408, 593–596 (2000)

    ADS  CAS  Article  Google Scholar 

  7. 7

    Peng, C. Y., Manning, L., Albertson, R. & Doe, C. Q. The tumour-suppressor genes lgl and dlg regulate basal protein targeting in Drosophila neuroblasts. Nature 408, 596–600 (2000)

    ADS  CAS  Article  Google Scholar 

  8. 8

    Albertson, R. & Doe, C. Q. Dlg, Scrib and Lgl regulate neuroblast cell size and mitotic spindle asymmetry. Nature Cell Biol. 5, 166–170 (2003)

    CAS  Article  Google Scholar 

  9. 9

    Strand, D. et al. The Drosophila lethal(2)giant larvae tumour suppressor protein forms homo-oligomers and is associated with nonmuscle myosin II heavy chain. J. Cell Biol. 127, 1361–1373 (1994)

    CAS  Article  Google Scholar 

  10. 10

    Betschinger, J., Mechtler, K. & Knoblich, J. A. The Par complex directs asymmetric cell division by phosphorylating the cytoskeletal protein Lgl. Nature 422, 326–330 (2003)

    ADS  CAS  Article  Google Scholar 

  11. 11

    Buenzow, D. E. & Holmgren, R. Expression of the Drosophila gooseberry locus defines a subset of neuroblast lineages in the central nervous system. Dev. Biol. 170, 338–349 (1995)

    CAS  Article  Google Scholar 

  12. 12

    Freeman, M. R. & Doe, C. Q. Asymmetric Prospero localization is required to generate mixed neuronal/glial lineages in the Drosophila CNS. Development 128, 4103–4112 (2001)

    CAS  PubMed  Google Scholar 

  13. 13

    Spana, E. P. & Doe, C. Q. The Prospero transcription factor is asymmetrically localized to the cell cortex during neuroblast mitosis in Drosophila. Development 121, 3187–3195 (1995)

    CAS  PubMed  Google Scholar 

  14. 14

    Bilder, D., Li, M. & Perrimon, N. Cooperative regulation of cell polarity and growth by Drosophila tumour suppressors. Science 289, 113–116 (2000)

    ADS  CAS  Article  Google Scholar 

  15. 15

    Hutterer, A., Betschinger, J., Petronczki, M. & Knoblich, J. A. Sequential roles of Cdc42, Par-6, aPKC, and Lgl in the establishment of epithelial polarity during Drosophila embryogenesis. Dev. Cell 6, 845–854 (2004)

    CAS  Article  Google Scholar 

  16. 16

    Yamanaka, T. et al. Mammalian Lgl forms a protein complex with PAR-6 and aPKC independently of PAR-3 to regulate epithelial cell polarity. Curr. Biol. 13, 734–743 (2003)

    CAS  Article  Google Scholar 

  17. 17

    Chalmers, A. D. et al. aPKC, Crumbs3 and Lgl2 control apicobasal polarity in early vertebrate development. Development 132, 977–986 (2005)

    CAS  Article  Google Scholar 

  18. 18

    Rolls, M. M., Albertson, R., Shih, H. P., Lee, C. Y. & Doe, C. Q. Drosophila aPKC regulates cell polarity and cell proliferation in neuroblasts and epithelia. J. Cell Biol. 163, 1089–1098 (2003)

    CAS  Article  Google Scholar 

  19. 19

    Sotillos, S., Diaz-Meco, M. T., Caminero, E., Moscat, J. & Campuzano, S. DaPKC-dependent phosphorylation of Crumbs is required for epithelial cell polarity in Drosophila. J. Cell Biol. 166, 549–557 (2004)

    CAS  Article  Google Scholar 

  20. 20

    Caussinus, E. & Gonzalez, C. Induction of tumour growth by altered stem-cell asymmetric division in Drosophila melanogaster. Nature Genet. 37, 1125–1129 (2005)

    CAS  Article  Google Scholar 

  21. 21

    Klezovitch, O., Fernandez, T. E., Tapscott, S. J. & Vasioukhin, V. Loss of cell polarity causes severe brain dysplasia in Lgl1 knockout mice. Genes Dev. 18, 559–571 (2004)

    CAS  Article  Google Scholar 

  22. 22

    Sanada, K. & Tsai, L. H. G Protein βγ subunits and AGS3 control spindle orientation and asymmetric cell fate of cerebral cortical progenitors. Cell 122, 119–131 (2005)

    CAS  Article  Google Scholar 

  23. 23

    Yu, F., Morin, X., Cai, Y., Yang, X. & Chia, W. Analysis of partner of inscuteable, a novel player of Drosophila asymmetric divisions, reveals two distinct steps in inscuteable apical localization. Cell 100, 399–409 (2000)

    CAS  Article  Google Scholar 

  24. 24

    Albertson, R., Chabu, C., Sheehan, A. & Doe, C. Q. Scribble protein domain mapping reveals a multistep localization mechanism and domains necessary for establishing cortical polarity. J. Cell Sci. 117, 6061–6070 (2004)

    CAS  Article  Google Scholar 

  25. 25

    Pearson, B. J. & Doe, C. Q. Regulation of neuroblast competence in Drosophila. Nature 425, 624–628 (2003)

    ADS  CAS  Article  Google Scholar 

  26. 26

    Freeman, M. R., Delrow, J., Kim, J., Johnson, E. & Doe, C. Q. Unwrapping glial biology: Gcm target genes regulating glial development, diversification, and function. Neuron 38, 567–580 (2003)

    CAS  Article  Google Scholar 

  27. 27

    Grosskortenhaus, R., Pearson, B. J., Marusich, A. & Doe, C. Q. Regulation of temporal identity transitions in Drosophila neuroblasts. Dev. Cell 8, 193–202 (2005)

    CAS  Article  Google Scholar 

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We thank J. Knoblich, S. Campuzano, B. Chia, J. Skeath and B. Holmgren for fly stocks and/or antibody reagents; B. Bowerman, J. Eisen, K. Siller and S. Siegrist for comments on the manuscript; and C. Chabu for discussion. C.-Y.L. is supported by a Damon Runynon postdoctoral fellowship. C.Q.D. is supported by the Howard Hughes Medical Institute, where he is an Investigator.

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Correspondence to Chris Q. Doe.

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Supplementary information

Supplementary Figure 1

lgl and pins regulate larval neuroblast numbers. (PDF 132 kb)

Supplementary Figure 2

Apoptosis is not increased in pins mutant brains. (PDF 59 kb)

Supplementary Figure 3

A comparison of neuroblast and GMC markers reveals that lgl pins mutant brains have supernumerary neuroblasts. (PDF 326 kb)

Supplementary Figure 4

aPKC is required for neuroblast self-renewal in lgl mutants. (PDF 120 kb)

Supplementary Data

Gene/protein list (DOC 19 kb)

Supplementary Methods

Additional description of the methods used in this study. (DOC 23 kb)

Supplementary Movie1

Thousands of differentiating GMCs and neurons rapidly down-regulate neuroblast markers and express nuclear Prospero and/or Elav. (MOV 2660 kb)

Supplementary Movie 2

There was a clear increase in neuroblast number in lgl and dlg mutants; and also supernumerary neuroblasts at all stages examined. All extra neuroblasts expressed Deadpan and Miranda neuroblast markers and were proliferative based on their ability to incorporate BrdU. (MOV 2750 kb)

Supplementary Movie3

Gαi zygotic mutants had a complex phenotype, but pins zygotic mutants showed a striking decrease in neuroblast number. (MOV 2450 kb)

Supplementary Movie 4

A novel phenotype was detected, in which the larval brain was full of cells expressing the neuroblast markers Worniu, Miranda and Deadpan, and lacking expression of the neuronal marker Elav. (MOV 2209 kb)

Supplementary Movie 5

Neuroblast-specific expression of aPKC targeted to the plasma membrane with a CAAX prenylation motif (UAS-aPKCCAAXWT) resulted in ectopic cortical aPKC localization, loss of cortical Miranda and a massive increase in the number of neuroblasts. (MOV 2686 kb)

Supplementary Movie 6

Neuroblast-specific expression of aPKC targeted to the plasma membrane with a CAAX prenylation motif (UAS-aPKCCAAXWT) resulted in ectopic cortical aPKC localization, loss of cortical Miranda and a massive increase in the number of neuroblasts. These effects were not observed following overexpression of wild type aPKC or a membrane-targeted kinase-dead aPKC (UAS-aPKCCAAXKD). (MOV 2431 kb)

Supplementary Figure Legends

Text to accompany the above Supplementary Figures. (DOC 21 kb)

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Lee, C., Robinson, K. & Doe, C. Lgl, Pins and aPKC regulate neuroblast self-renewal versus differentiation. Nature 439, 594–598 (2006). https://doi.org/10.1038/nature04299

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