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J Vis Exp. 2014 Jul 13;(89). doi: 10.3791/51634.

A method of permeabilization of Drosophila embryos for assays of small molecule activity.

Author information

1
Department of Environmental Medicine, University of Rochester School of Dentistry and Medicine; Matthew_Rand@urmc.rochester.edu.

Abstract

The Drosophila embryo has long been a powerful laboratory model for elucidating molecular and genetic mechanisms that control development. The ease of genetic manipulations with this model has supplanted pharmacological approaches that are commonplace in other animal models and cell-based assays. Here we describe recent advances in a protocol that enables application of small molecules to the developing fruit fly embryo. The method details steps to overcome the impermeability of the eggshell while maintaining embryo viability. Eggshell permeabilization across a broad range of developmental stages is achieved by application of a previously described d-limonene embryo permeabilization solvent (EPS1) and by aging embryos at reduced temperature (18 °C) prior to treatments. In addition, use of a far-red dye (CY5) as a permeabilization indicator is described, which is compatible with downstream applications involving standard red and green fluorescent dyes in live and fixed preparations. This protocol is applicable to studies using bioactive compounds to probe developmental mechanisms as well as for studies aimed at evaluating teratogenic or pharmacologic activity of uncharacterized small molecules.

PMID:
25046169
PMCID:
PMC4214599
DOI:
10.3791/51634
[Indexed for MEDLINE]
Free PMC Article

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