Germline Clone Analysis for Maternally Acting Drosophila Hedgehog Components
Many of the genes of Drosophila melanogaster have their transcripts deposited in developing oocytes. These maternally loaded gene products enable an otherwise homozygous mutant embryo to survive beyond the first stage of development for which the gene product is required. Zygotic mutations that disrupt the Hedgehog signal transduction pathway typically yield a segment polarity ‘lawn of denticles’ cuticle phenotype. However, an embryo homozygous mutant for a gene can achieve normal embryonic segmentation precluding classification of the gene as a component of the Hh pathway, if wild-type transcripts from the mother are present. This chapter discusses the theory and importance of analyzing germline clone embryos for maternally acting genes involved in Hh signal transduction, and describes in detail the method to generate mutant germline clone embryos.
Key WordsGermline clone embryos germline stem cells Hh signal transduction recombination FLP recombinase maternal transcripts Drosophila
- 10.Martinez-Arias, A., Baker, N., and Ingham, P. W. (1988) Role of segment polarity genes in the definition and maintenance of cell states in Drosophila embryo. Development 103, 157–170.Google Scholar
- 13.Rothwell, F. and Sullivan, W. (2000) Fluorescent analysis of Drosophila embryos. In Drosophila Protocols, Cold Spring Harbor Laboratory Press, New York, pp. 141–157.Google Scholar
- 15.Parthasarathy, N., Lecuyer, E., and Krause, H. M. (2005) Optimized protocols for fluorescent in situ hybridization in Drosophila tissues. In Methods in Molecular Biology: Protocols in Confocal Microscopy (Paddock, S., ed.) Humana Press Inc.Google Scholar