Skip to main content
SpringerLink
Log in

Transcriptome Analysis of Drosophila Neural Stem Cells

  • Protocol
  • First Online:
Progenitor Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 916))

  • 3089 Accesses

Abstract

In Drosophila, the central nervous system is populated by a set of asymmetrically dividing neural stem cells called neuroblasts. Neuroblasts are derived from epithelial or neuroepithelial precursors, and divide along their apico-basal axes to produce a large apical neuroblast and a smaller basal ganglion mother cell. The ganglion mother cell will divide once again to produce two post-mitotic neurons or glia. In this chapter we outline a method for labeling different types of neural precursors in the Drosophila central nervous system, followed by their extraction and processing for transcriptome analysis. This technique has allowed us to capture and compare the expression profiles of neuroblasts and neuroepithelial cells, resulting in the identification of key genes required for the regulation of self-renewal and differentiation.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Technau GM, Campos-Ortega JA (1985) Fate-mapping in wild-type Drosophila melanogaster. Dev Genes Evol 194:196–212

    Google Scholar 

  2. Technau GM (1987) A single cell approach to problems of cell lineage and commitment during embryogenesis of Drosophila melanogaster. Development 100:1–12

    PubMed  CAS  Google Scholar 

  3. Bossing T, Technau GM (1994) The fate of the CNS midline progenitors in Drosophila as revealed by a new method for single cell labelling. Development 120:1895–1906

    PubMed  CAS  Google Scholar 

  4. Lee T, Luo L (1999) Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis. Neuron 22:451–461

    Article  PubMed  CAS  Google Scholar 

  5. Evans CJ, Olson JM, Ngo KT, Kim E, Lee NE, Kuoy E, Patananan AN, Sitz D, Tran P, Do MT, Yackle K, Cespedes A, Hartenstein V, Call GB, Banerjee U (2009) G-TRACE: rapid Gal4-based cell lineage analysis in Drosophila. Nat Methods 6:603–605

    Article  PubMed  CAS  Google Scholar 

  6. Griffin R, Sustar A, Bonvin M, Binari R, del Valle Rodriguez A, Hohl AM, Bateman JR, Villalta C, Heffern E, Grunwald D, Bakal C, Desplan C, Schubiger G, Wu CT, Perrimon N (2009) The twin spot generator for differential Drosophila lineage analysis. Nat Methods 6:600–602

    Article  PubMed  CAS  Google Scholar 

  7. Yu HH, Chen CH, Shi L, Huang Y, Lee T (2009) Twin-spot MARCM to reveal the developmental origin and identity of neurons. Nat Neurosci 12:947–953

    Article  PubMed  CAS  Google Scholar 

  8. Lee T (2009) New genetic tools for cell lineage analysis in Drosophila. Nat Methods 6:566–568

    Article  PubMed  CAS  Google Scholar 

  9. Doe CQ (1992) Molecular markers for identified neuroblasts and ganglion mother cells in the Drosophila central nervous system. Development 116:855–863

    PubMed  CAS  Google Scholar 

  10. Broadus J, Skeath JB, Spana EP, Bossing T, Technau G, Doe CQ (1995) New neuroblast markers and the origin of the aCC/pCC neurons in the Drosophila central nervous system. Mech Dev 53:393–402

    Article  PubMed  CAS  Google Scholar 

  11. Bossing T, Udolph G, Doe CQ, Technau GM (1996) The embryonic central nervous system lineages of Drosophila melanogaster. I. Neuroblast lineages derived from the ventral half of the neuroectoderm. Dev Biol 179:41–64

    Article  PubMed  CAS  Google Scholar 

  12. Schmidt H, Rickert C, Bossing T, Vef O, Urban J, Technau GM (1997) The embryonic central nervous system lineages of Drosophila melanogaster. II. Neuroblast lineages derived from the dorsal part of the neuroectoderm. Dev Biol 189:186–204

    Article  PubMed  CAS  Google Scholar 

  13. Spindler SR, Hartenstein V (2010) The Drosophila neural lineages: a model system to study brain development and circuitry. Dev Genes Evol 220:1–10

    Article  PubMed  Google Scholar 

  14. Urbach R, Schnabel R, Technau GM (2003) The pattern of neuroblast formation, mitotic domains and proneural gene expression during early brain development in Drosophila. Development 130:3589–3606

    Article  PubMed  CAS  Google Scholar 

  15. Urbach R, Technau GM (2003) Segment polarity and DV patterning gene expression reveals segmental organization of the Drosophila brain. Development 130:3607–3620

    Article  PubMed  CAS  Google Scholar 

  16. Urbach R, Technau GM (2003) Molecular markers for identified neuroblasts in the developing brain of Drosophila. Development 130:3621–3637

    Article  PubMed  CAS  Google Scholar 

  17. Megason SG, Fraser SE (2003) Digitizing life at the level of the cell: high-performance laser-scanning microscopy and image analysis for in to imaging of development. Mech Dev 120:1407–1420

    Article  PubMed  CAS  Google Scholar 

  18. Walter T, Shattuck DW, Baldock R, Bastin ME, Carpenter AE, Duce S, Ellenberg J, Fraser A, Hamilton N, Pieper S, Ragan MA, Schneider JE, Tomancak P, Heriche JK (2010) Visualization of image data from cells to organisms. Nat Methods 7:S26–41

    Article  PubMed  CAS  Google Scholar 

  19. Southall TD, Egger B, Gold KS, Brand AH (2008) Regulation of self-renewal and differentiation in the Drosophila nervous system. Cold Spring Harb Symp Quant Biol 73:523–528

    Article  PubMed  CAS  Google Scholar 

  20. Egger B, Chell JM, Brand AH (2008) Insights into neural stem cell biology from flies. Phil Trans R Soc Lond 363:39–56

    Article  CAS  Google Scholar 

  21. Doe CQ (2008) Neural stem cells: balancing self-renewal with differentiation. Development 135:1575–1587

    Article  PubMed  CAS  Google Scholar 

  22. Choksi SP, Southall TD, Bossing T, Edoff K, de Wit E, Fischer BE, van Steensel B, Micklem G, Brand AH (2006) Prospero acts as a binary switch between self-renewal and differentiation in Drosophila neural stem cells. Dev Cell 11:775–789

    Article  PubMed  CAS  Google Scholar 

  23. Egger B, Gold KS, Brand AH (2010) Notch regulates the switch from symmetric to asymmetric neural stem cell division in the Drosophila optic lobe. Development 137:2981–2987

    Article  PubMed  CAS  Google Scholar 

  24. Technau GM (1986) Lineage analysis of transplanted individual cells in embryos of Drosophila melanogaster. Dev Genes Evol 195:389–398

    Google Scholar 

  25. Prokop A, Technau GM (1993) Cell transplantation. In: Hartley D (ed) Cellular interactions in development: a practical approach. Oxford University Press, p 33–57

    Google Scholar 

  26. Iscove NN, Barbara M, Gu M, Gibson M, Modi C, Winegarden N (2002) Representation is faithfully preserved in global cDNA amplified exponentially from sub-picogram quantities of mRNA. Nat Biotechnol 20:940–943

    Article  PubMed  CAS  Google Scholar 

  27. Osawa M, Egawa G, Mak SS, Moriyama M, Freter R, Yonetani S, Beermann F, Nishikawa S (2005) Molecular characterization of melanocyte stem cells in their niche. Development 132:5589–5599

    Article  PubMed  CAS  Google Scholar 

  28. Saito H, Kubota M, Roberts RW, Chi Q, Matsunami H (2004) RTP family members induce functional expression of mammalian odorant receptors. Cell 119:679–691

    Article  PubMed  CAS  Google Scholar 

  29. Subkhankulova T, Livesey FJ (2006) Comparative evaluation of linear and exponential amplification techniques for expression profiling at the single-cell level. Genome Biol 7:R18

    Article  PubMed  Google Scholar 

  30. Egger B, Boone JQ, Stevens NR, Brand AH, Doe CQ (2007) Regulation of spindle orientation and neural stem cell fate in the Drosophila optic lobe. Neural Dev 2:1

    Article  PubMed  Google Scholar 

  31. Hofbauer A, Campos-Ortega JA (1990) Proliferation pattern and early differentiation of the optic lobes in Drosophila melanogaster. Dev Genes Evol 198:264–274

    Google Scholar 

  32. Pfeiffer BD, Jenett A, Hammonds AS, Ngo TT, Misra S, Murphy C, Scully A, Carlson JW, Wan KH, Laverty TR, Mungall C, Svirskas R, Kadonaga JT, Doe CQ, Eisen MB, Celniker SE, Rubin GM (2008) Tools for neuroanatomy and neurogenetics in Drosophila. Proc Natl Acad Sci USA 105:9715–9720

    Article  PubMed  CAS  Google Scholar 

  33. Pfeiffer BD, Ngo TT, Hibbard KL, Murphy C, Jenett A, Truman JW, Rubin GM (2010) Refinement of tools for targeted gene expression in Drosophila. Genetics 186:735–755

    Article  PubMed  CAS  Google Scholar 

  34. Potter CJ, Tasic B, Russler EV, Liang L, Luo L (2010) The Q system: a repressible binary system for transgene expression, lineage tracing, and mosaic analysis. Cell 141:536–548

    Article  PubMed  CAS  Google Scholar 

  35. Diegelmann S, Bate M, Landgraf M (2008) Gateway cloning vectors for the LexA-based binary expression system in Drosophila. Fly 2:236–239

    PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Karin Edoff, Boris Egger, Paul Wu, Adrian Carr, Hiroaki Matsunami, and Rick Livesey for sharing protocols and their considerable expertise. This work was supported by the Wellcome Trust.

Author information

Authors and Affiliations

  1. The Gurdon Institute, University of Cambridge, Cambridge, UK

    Katrina S. Gold & Andrea H. Brand

Authors
  1. Katrina S. Gold
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andrea H. Brand
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Andrea H. Brand .

Editor information

Editors and Affiliations

  1. Fac. Life Sciences, Healing Foundation Centre, University of Manchester, Oxford Road, Manchester, M13 9PT, United Kingdom

    Kimberly A. Mace

  2. School of Medicine & Dentistry, Centre for Cutaneous Research, Blizard I, Barts and The London, Newark Street 4, London, E1 2AT, United Kingdom

    Kristin M. Braun

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media New York

About this protocol

Cite this protocol

Gold, K.S., Brand, A.H. (2012). Transcriptome Analysis of Drosophila Neural Stem Cells. In: Mace, K., Braun, K. (eds) Progenitor Cells. Methods in Molecular Biology, vol 916. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-980-8_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-980-8_8

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-979-2

  • Online ISBN: 978-1-61779-980-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation