Abstract
Phosphoinositides (PIs), the seven phosphorylated derivatives of phosphatidylinositol, are recognized as key molecules in the control of multiple molecular events in eukaryotic cells. Within cells, PIs are low-abundance lipids making their detection and quantification challenging. While many methods that allow radiolabeling and quantification of PIs in the context of cultured cells are available, these are not useful in the context of in vivo animal models where cell and developmental processes are best studied. In this chapter, we describe radionuclide-free, mass spectrometry-based methods for the detection and quantification of PIs from Drosophila tissues in vivo. The use of these methods should facilitate the discovery of novel modes by which PIs regulate cellular and developmental processes in complex metazoans.
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Acknowledgments
This work was supported by the National Centre for Biological Sciences-TIFR and a Wellcome DBT India Alliance Senior Fellowship to PR (IA/S/14/2/501540). We thank Dhananjay Shinde, Sanjeev Sharma, Sruthi Balakrishnan, and the NCBS mass spectrometry Facility for assistance while developing these methods.
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Ghosh, A., Raghu, P. (2021). Label-Free Quantification of Phosphoinositides in Drosophila by Mass Spectrometry. In: Botelho, R.J. (eds) Phosphoinositides. Methods in Molecular Biology, vol 2251. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1142-5_2
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DOI: https://doi.org/10.1007/978-1-0716-1142-5_2
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