Abstract
Genetic mosaic analyses represent an invaluable approach for the study of stem cell lineages in the Drosophila ovary. The generation of readily identifiable, homozygous mutant cells in the context of wild-type ovarian tissues within intact organisms allows the pinpointing of cellular requirements for gene function, which is particularly important for understanding the physiological control of stem cells and their progeny. Here, we provide a step-by-step guide to the generation and analysis of genetically mosaic ovaries using flippase (FLP)/FLP recognition target (FRT)-mediated recombination in adult Drosophila melanogaster, with a focus on the processes of oogenesis that are controlled by diet-dependent factors.
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References
Perrimon N (1998) Creating mosaics in Drosophila. Int J Dev Biol 42:243–247
Theodosiou NA, Xu T (1998) Use of FLP/FRT system to study Drosophila development. Methods 14:355–365
Le Lievre CS, Le Douarin NM (1975) Mesenchymal derivatives of the neural crest: analysis of chimaeric quail and chick embryos. J Embryol Exp Morphol 34:125–154
Lehmann R, Nusslein-Volhard C (1987) hunchback, a gene required for segmentation of an anterior and posterior region of the Drosophila embryo. Dev Biol 119:402–417
Hadorn E (1968) Transdetermination in cells. Sci Am 219:110–114 passim
Simon MA, Bowtell DD, Dodson GS et al (1991) Ras1 and a putative guanine nucleotide exchange factor perform crucial steps in signaling by the sevenless protein tyrosine kinase. Cell 67:701–716
Xu T, Rubin GM (1993) Analysis of genetic mosaics in developing and adult Drosophila tissues. Development 117:1223–1237
Lee T, Luo L (2001) Mosaic analysis with a repressible cell marker (MARCM) for Drosophila neural development. Trends Neurosci 24:251–254
Evans CJ, Olson JM, Ngo KT et al (2009) G-TRACE: rapid Gal4-based cell lineage analysis in Drosophila. Nat Methods 6:603–605
Struhl G, Basler K (1993) Organizing activity of wingless protein in Drosophila. Cell 72:527–540
Ables ET, Drummond-Barbosa D (2010) The steroid hormone ecdysone functions with intrinsic chromatin remodeling factors to control female germline stem cells in Drosophila. Cell Stem Cell 7:581–592
Ables ET, Drummond-Barbosa D (2013) Cyclin E controls Drosophila female germline stem cell maintenance independently of its role in proliferation by modulating responsiveness to niche signals. Development 140:530–540
Hsu HJ, Drummond-Barbosa D (2009) Insulin levels control female germline stem cell maintenance via the niche in Drosophila. Proc Natl Acad Sci U S A 106:1117–1121
Hsu HJ, Drummond-Barbosa D (2011) Insulin signals control the competence of the Drosophila female germline stem cell niche to respond to Notch ligands. Dev Biol 350:290–300
Hsu HJ, LaFever L, Drummond-Barbosa D (2008) Diet controls normal and tumorous germline stem cells via insulin-dependent and -independent mechanisms in Drosophila. Dev Biol 313:700–712
LaFever L, Drummond-Barbosa D (2005) Direct control of germline stem cell division and cyst growth by neural insulin in Drosophila. Science 309:1071–1073
LaFever L, Feoktistov A, Hsu HJ et al (2010) Specific roles of Target of rapamycin in the control of stem cells and their progeny in the Drosophila ovary. Development 137:2117–2126
Margolis J, Spradling A (1995) Identification and behavior of epithelial stem cells in the Drosophila ovary. Development 121:3797–3807
Spradling AC (1993) Developmental genetics of oogenesis. In: Bate M (ed) The development of Drosophila melanogaster. Cold Spring Harbor Laboratory Press, Plainview, NY
Haack T, Bergstralh DT, St Johnston D (2013) Damage to the Drosophila follicle cell epithelium produces “false clones” with apparent polarity phenotypes. Biol Open 2:1313–1320
Cummings MR, Brown NM, King RC (1971) The cytology of the vitellogenic stages of oogenesis in Drosophila melanogaster. 3. Formation of the vitelline membrane. Z Zellforsch Mikrosk Anat 118:482–492
Xie T, Spradling AC (2000) A niche maintaining germ line stem cells in the Drosophila ovary. Science 290:328–330
Sahai-Hernandez P, Castanieto A, Nystul TG (2012) Drosophila models of epithelial stem cells and their niches. Wiley Interdiscip Rev Dev Biol 1:447–457
de Cuevas M, Spradling AC (1998) Morphogenesis of the Drosophila fusome and its implications for oocyte specification. Development 125:2781–2789
Acknowledgements
We are grateful to members of the Drummond-Barbosa lab for critical comments during the preparation of this manuscript. This work was supported by National Institutes of Health (NIH) grant R01 GM069875 (D.D.B.). K.L. was supported by NIH training grant T32CA009110.
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Laws, K.M., Drummond-Barbosa, D. (2015). Genetic Mosaic Analysis of Stem Cell Lineages in the Drosophila Ovary. 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_4
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DOI: https://doi.org/10.1007/978-1-4939-2851-4_4
Publisher Name: Humana Press, New York, NY
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