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A spindle-independent cleavage pathway controls germ cell formation in Drosophila

Nature Cell Biology volume 15, pages 839–845 (2013)Cite this article

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Abstract

The primordial germ cells (PGCs) are the first cells to form during Drosophila melanogaster embryogenesis. Whereas the process of somatic cell formation has been studied in detail, the mechanics of PGC formation are poorly understood. Here, using four-dimensional multi-photon imaging combined with genetic and pharmacological manipulations, we find that PGC formation requires an anaphase spindle-independent cleavage pathway. In addition to using core regulators of cleavage, including the small GTPase RhoA (Drosophila rho1) and the Rho-associated kinase, ROCK (Drosophila drok), we show that this pathway requires Germ cell-less (GCL), a conserved BTB-domain protein not previously implicated in cleavage mechanics. This alternative form of cell formation suggests that organisms have evolved multiple molecular strategies for regulating the cytoskeleton during cleavage.

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Figure 1: Anillin–GFP and Myosin–GFP localize to paired cleavage furrows during Drosophila PGC formation.
Figure 2: A spindle-independent cleavage pathway directs BF cleavage.
Figure 3: Germ cell-less is required for BF constriction.
Figure 4: Germ cell-less is a rate-limiting component of BF constriction.
Figure 5: Mis-expression of Germ cell-less with Anillin is sufficient for ectopic cell formation.

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Acknowledgements

We thank all members of the Lehmann laboratory for discussions and reagents. We thank A. Blum, S. Burden, T. Hurd, M. Slaidina, F. Teixeira and A. Zamparini for critical reading of the manuscript. We also thank S. Burden, H. Knaut, J. Nance, G. Schubiger, S. Small, J. Treisman and E. Wieschaus for discussions during the course of this work. We thank the Drosophila Bloomington Stock Center, T. Jongens, C. Fields, H. Sano, and A. Wilde for reagents. This work was supported by a NSF Predoctoral Fellowship to R.M.C. R.L. is a Howard Hughes Medical Institute Investigator. We dedicate this manuscript to the memory of G. Schubiger whose work in the early Drosophila embryo largely inspired the experiments presented here.

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Authors and Affiliations

  1. Department of Cell Biology, HHMI and Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, New York 10016, USA

    Ryan M. Cinalli & Ruth Lehmann

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  1. Ryan M. Cinalli
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Contributions

R.M.C and R.L. conceived the project. R.M.C carried out the experiments and analysed the data. R.M.C and R.L. wrote the paper.

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Correspondence to Ruth Lehmann.

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Supplementary information

Supplementary Information

Supplementary Information (PDF 441 kb)

PGC formation in an embryo expressing Anillin–GFP.

This video shows four paired furrows forming and then constricting. (MOV 158 kb)

Paired furrow constriction in a Myosin–GFP and Vasa-KO embryo.

This video shows a single paired furrow constricting. Myosin–GFP (green) and Vasa-KO (red). (MOV 69 kb)

Anillin–GFP expression at the BF following DMSO injection.

DMSO injection does not disrupt the localization of Anillin at the BF. (MOV 20 kb)

Anillin–GFP expression at the BF following C3 peptide injection.

C3 peptide injection does disrupt the localization of Anillin at the BF. (MOV 31 kb)

BF constriction following colcemid injection in an embryo expressing Anillin–GFP.

Note that although the BF constricts, the AF does not form. (MOV 54 kb)

PGC formation in a gcl mutant; embryo expressing Anillin–GFP.

This video shows that the BF fails to constrict in gcl mutants. (MOV 159 kb)

BF constriction following colcemid injection in an embryo expressing Anillin–GFP.

Note that although the BF constricts, the AF does not form. (MOV 69 kb)

BF constriction following colcemid injection in a gcl mutant; embryo expressing Anillin–GFP.

Note that the BF fails to constrict. (MOV 78 kb)

BF constriction following in an embryo expressing Anillin–GFP. (MOV 11 kb)

BF constriction in a gcl mutant; embryo expressing Anillin–GFP. (MOV 9 kb)

BF constriction following colcemid injection in an embryo over-expressing gcl and expressing Anillin–GFP. (MOV 11 kb)

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Cinalli, R., Lehmann, R. A spindle-independent cleavage pathway controls germ cell formation in Drosophila. Nat Cell Biol 15, 839–845 (2013). https://doi.org/10.1038/ncb2761

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