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Analysis of Cell Cycle Switches in Drosophila Oogenesis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1328))

Abstract

The study of Drosophila oogenesis provides invaluable information about signaling pathway regulation and cell cycle programming. During Drosophila oogenesis, a string of egg chambers in each ovariole progressively develops toward maturity. Egg chamber development consists of 14 stages. From stage 1 to stage 6 (mitotic cycle), main-body follicle cells undergo mitotic divisions. From stage 7 to stage 10a (endocycle), follicle cells cease mitosis but continue three rounds of endoreduplication. From stage 10b to stage 13 (gene amplification), instead of whole genome duplication, follicle cells selectively amplify specific genomic regions, mostly for chorion production. So far, Drosophila oogenesis is one of the most well studied model systems used to understand cell cycle switches, which furthers our knowledge about cell cycle control machinery and sheds new light on potential cancer treatments. Here, we give a brief summary of cell cycle switches, the associated signaling pathways and factors, and the detailed experimental procedures used to study the cell cycle switches.

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References

  1. Spradling A (1993) Developmental genetics of oogenesis. The development of Drosophila melanogaster. Cold Spring Harbor Lab Press, New York

    Google Scholar 

  2. Nystul T, Spradling A (2010) Regulation of epithelial stem cell replacement and follicle formation in the Drosophila ovary. Genetics 184:503–515

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  3. Skora AD, Spradling AC (2010) Epigenetic stability increases extensively during Drosophila follicle stem cell differentiation. Proc Natl Acad Sci U S A 107:7389–7394

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  4. Calvi BR, Lilly MA, Spradling AC (1998) Cell cycle control of chorion gene amplification. Genes Dev 12:734–744

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Sun J, Deng WM (2005) Notch-dependent downregulation of the homeodomain gene cut is required for the mitotic cycle/endocycle switch and cell differentiation in Drosophila follicle cells. Development 132:4299–4308

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  6. Sun J, Deng WM (2007) Hindsight mediates the role of notch in suppressing hedgehog signaling and cell proliferation. Dev Cell 12:431–442

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Shyu LF, Sun J, Chung HM et al (2009) Notch signaling and developmental cell-cycle arrest in Drosophila polar follicle cells. Mol Biol Cell 20:5064–5073

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Ruohola H, Bremer KA, Baker D et al (1991) Role of neurogenic genes in establishment of follicle cell fate and oocyte polarity during oogenesis in Drosophila. Cell 66:433–449

    Article  CAS  PubMed  Google Scholar 

  9. Xu T, Caron LA, Fehon RG et al (1992) The involvement of the Notch locus in Drosophila oogenesis. Development 115:913–922

    CAS  PubMed  Google Scholar 

  10. Deng WM, Althauser C, Ruohola-Baker H (2001) Notch-Delta signaling induces a transition from mitotic cell cycle to endocycle in Drosophila follicle cells. Development 128:4737–4746

    CAS  PubMed  Google Scholar 

  11. Lopez-Schier H, St Johnston D (2001) Delta signaling from the germ line controls the proliferation and differentiation of the somatic follicle cells during Drosophila oogenesis. Genes Dev 15:1393–1405

    Article  CAS  PubMed  Google Scholar 

  12. Sun J, Smith L, Armento A et al (2008) Regulation of the endocycle/gene amplification switch by Notch and ecdysone signaling. J Cell Biol 182:885–896

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Yu J, Poulton J, Huang YC et al (2008) The hippo pathway promotes Notch signaling in regulation of cell differentiation, proliferation, and oocyte polarity. PLoS One 3:e1761

    Article  PubMed Central  PubMed  Google Scholar 

  14. Axelrod JD, Matsuno K, Artavanis-Tsakonas S et al (1996) Interaction between Wingless and Notch signaling pathways mediated by dishevelled. Science 271:1826–1832

    Article  CAS  PubMed  Google Scholar 

  15. Jordan KC, Schaeffer V, Fischer KA et al (2006) Notch signaling through tramtrack bypasses the mitosis promoting activity of the JNK pathway in the mitotic-to-endocycle transition of Drosophila follicle cells. BMC Dev Biol 6:16

    Article  PubMed Central  PubMed  Google Scholar 

  16. White AE, Leslie ME, Calvi BR et al (2007) Developmental and cell cycle regulation of the Drosophila histone locus body. Mol Biol Cell 18:2491–2502

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Royzman I, Austin RJ, Bosco G et al (1999) ORC localization in Drosophila follicle cells and the effects of mutations in dE2F and dDP. Genes Dev 13:827–840

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Zielke N, Korzelius J, van Straaten M et al (2014) Fly-FUCCI: A versatile tool for studying cell proliferation in complex tissues. Cell Rep 7:588–598

    Article  CAS  PubMed  Google Scholar 

  19. Jia D, Tamori Y, Pyrowolakis G et al (2014) Regulation of broad by the Notch pathway affects timing of follicle cell development. Dev Biol 392:52–61

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  20. de Celis JF, Tyler DM, de Celis J et al (1998) Notch signalling mediates segmentation of the Drosophila leg. Development 125:4617–4626

    PubMed  Google Scholar 

  21. Assa-Kunik E, Torres IL, Schejter ED et al (2007) Drosophila follicle cells are patterned by multiple levels of Notch signaling and antagonism between the Notch and JAK/STAT pathways. Development 134:1161–1169

    Article  CAS  PubMed  Google Scholar 

  22. Furriols M, Bray S (2001) A model Notch response element detects Suppressor of Hairless-dependent molecular switch. Curr Biol 11:60–64

    Article  CAS  PubMed  Google Scholar 

  23. Saj A, Arziman Z, Stempfle D et al (2010) A combined ex vivo and in vivo RNAi screen for notch regulators in Drosophila reveals an extensive notch interaction network. Dev Cell 18:862–876

    Article  CAS  PubMed  Google Scholar 

  24. Fuchs A, Cheung LS, Charbonnier E et al (2012) Transcriptional interpretation of the EGF receptor signaling gradient. Proc Natl Acad Sci U S A 109:1572–1577

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  25. Wong LC, Schedl P (2006) Dissection of Drosophila ovaries. J Vis Exp 52

    Google Scholar 

  26. Bryant Z, Subrahmanyan L, Tworoger M et al (1999) Characterization of differentially expressed genes in purified Drosophila follicle cells: toward a general strategy for cell type-specific developmental analysis. Proc Natl Acad Sci U S A 96:5559–5564

    Article  PubMed Central  CAS  PubMed  Google Scholar 

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Acknowledgements

We would like to thank Gabriel Calvin, Gengqiang Xie, Allison Jevitt, and Sarayu Row for critical reading of and comments on the manuscript. Special thanks to Jianjun Sun and other former and current members of the Deng lab for developing protocols on this topic; W.-M.D. is supported by NIH grant R01GM072562.

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Correspondence to Wu-Min Deng .

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Jia, D., Huang, YC., Deng, WM. (2015). Analysis of Cell Cycle Switches in Drosophila Oogenesis. In: Bratu, D., McNeil, G. (eds) Drosophila Oogenesis. Methods in Molecular Biology, vol 1328. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2851-4_15

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  • DOI: https://doi.org/10.1007/978-1-4939-2851-4_15

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2850-7

  • Online ISBN: 978-1-4939-2851-4

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