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Broadie Laboratory

Structural mass spectrometry analysis of lipid changes in a Drosophila epilepsy model brain.

AUTHORS

Kliman MMichal , Vijayakrishnan N Niranjana , Wang L Lily , Tapp JT John T , Broadie K Kendal , McLean JA John A . Molecular bioSystems. 2010 6 ; 6(6). 958-66

ABSTRACT

Phosphatidylethanolamine (PtdEtn) is one of the most abundant phospholipids in many animal cell types. The Drosophila easily shocked (eas(2)) mutant, used as an epilepsy model, is null for the PtdEtn biosynthetic enzyme, ethanolamine kinase. This mutant displays bang sensitive paralysis, and was previously shown to have decreased levels of PtdEtn. We have developed a highly selective and sensitive measurement strategy using ion mobility-mass spectrometry for the relative quantitation of intact phospholipid species directly from isolated brain tissue of eas mutants. Over 1200 distinct lipid signals are observed and within this population 38, including PtdEtn, phosphatidylinositol (PtdIns) and phosphatidylcholine (PtdCho) species are identified to have changed significantly (p < 0.03) between mutant and control tissue. This method has revealed for the first time the structural complexity and biosynthetic interconnectedness of specific PtdEtn and PtdIns lipid species within tissue, and provides great molecular detail compared to traditionally used detection techniques.

Phosphatidylethanolamine (PtdEtn) is one of the most abundant phospholipids in many animal cell types. The Drosophila easily shocked (eas(2)) mutant, used as an epilepsy model, is null for the PtdEtn biosynthetic enzyme, ethanolamine kinase. This mutant displays bang sensitive paralysis, and was previously shown to have decreased levels of PtdEtn. We have developed a highly selective and sensitive measurement strategy using ion mobility-mass spectrometry for the relative quantitation of intact phospholipid species directly from isolated brain tissue of eas mutants. Over 1200 distinct lipid signals are observed and within this population 38, including PtdEtn, phosphatidylinositol (PtdIns) and phosphatidylcholine (PtdCho) species are identified to have changed significantly (p < 0.03) between mutant and control tissue. This method has revealed for the first time the structural complexity and biosynthetic interconnectedness of specific PtdEtn and PtdIns lipid species within tissue, and provides great molecular detail compared to traditionally used detection techniques.

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