Abstract
Fluorescent in situ hybridization (FISH) is a technique for determining the cytological localization of RNA or DNA molecules. There are many approaches available for generating in situ hybridization probes and conducting the subsequent hybridization steps. Here, we describe a simple and reliable FISH method to label small RNAs (200–500 nucleotides in length) that are enriched in nuclear bodies in Drosophila melanogaster ovaries, such as Cajal bodies (CBs) and histone locus bodies (HLBs). This technique can also be applied to other Drosophila tissues, and to abundant mRNAs such as histone transcripts.
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References
Nizami Z, Deryusheva S, Gall JG (2010) The Cajal body and histone locus body. Cold Spring Harb Perspect Biol 2:a000653.
Nizami ZF, Deryusheva S, Gall JG (2010) Cajal bodies and histone locus bodies in Drosophila and Xenopus. Cold Spring Harb Symp Quant Biol 75:313-320.
Liu J-L, Murphy C, Buszczak M et al. (2006) The Drosophila melanogaster Cajal body. J Cell Biol 172:875–884.
Liu J-L, Wu Z, Nizami Z et al. (2009) Coilin is essential for Cajal body organization in Drosophila melanogaster. Mol Biol Cell 20:1661–1670.
Deryusheva S, Gall JG (2013) Novel small Cajal-body-specific RNAs identified in Drosophila: probing guide RNA function. RNA 19:1802–1814.
Dej KJ, Spradling AC (1999) The endocycle controls nurse cell polytene chromosome structure during Drosophila oogenesis. Development 126:293–303.
Liu JL, Gall JG (2007) U bodies are cytoplasmic structures that contain uridine-rich small nuclear ribonucleoproteins and associate with P bodies. Proc Natl Acad Sci USA 104:11655–11659.
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.
Godfrey AC, Kupsco JM, Burch BD et al. (2006) U7 snRNA mutations in Drosophila block histone pre-mRNA processing and disrupt oogenesis. RNA 12:396–409.
Buckingham M, Liu JL (2011) U bodies respond to nutrient stress in Drosophila. Exp Cell Res 317:2835–2844.
Shimada Y, Burn KM, Niwa R et al. (2011) Reversible response of protein localization and microtubule organization to nutrient stress during Drosophila early oogenesis. Dev Biol 355:250–262.
Singer AB, Gall JG (2011) An inducible nuclear body in the Drosophila germinal vesicle. Nucleus 2:403–409.
Acknowledgements
We thank Svetlana Deryusheva (Carnegie Institution for Science) for useful advice and enhancements to this protocol.
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Nizami, Z.F., Liu, JL., Gall, J.G. (2015). Fluorescent In Situ Hybridization of Nuclear Bodies in Drosophila melanogaster Ovaries. 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_10
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DOI: https://doi.org/10.1007/978-1-4939-2851-4_10
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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