Studying fatty aldehyde metabolism in living cells with pyrene-labeled compounds

Markus A Keller,Katrin Watschinger,Karsten Lange,Georg Golderer,Gabriele Werner-Felmayer,Albin Hermetter,Ronald J.A Wanders,Ernst R Werner

doi:10.1194/jlr.d025650

Abstract

The lack of fatty aldehyde dehydrogenase function in Sjögren Larsson Syndrome (SLS) patient cells not only impairs the conversion of fatty aldehydes into their corresponding fatty acid but also has an effect on connected pathways. Alteration of the lipid profile in these cells is thought to be responsible for severe symptoms such as ichtyosis, mental retardation, and spasticity. Here we present a novel approach to examine fatty aldehyde metabolism in a time-dependent manner by measuring pyrene-labeled fatty aldehyde, fatty alcohol, fatty acid, and alkylglycerol in the culture medium of living cells using HPLC separation and fluorescence detection. Our results show that in fibroblasts from SLS patients, fatty aldehyde is not accumulating but is converted readily into fatty alcohol. In control cells, in contrast, exclusively the corresponding fatty acid is formed. SLS patient cells did not display a hypersensitivity toward hexadecanal or hexadecanol, but 3-fold lower concentrations of the fatty alcohol than the corresponding fatty aldehyde were needed to induce toxicity in SLS patient and in control cells.

Highlights

The lack of fatty aldehyde dehydrogenase function in Sjögren Larsson Syndrome (SLS) patient cells impairs the conversion of fatty aldehydes into their corresponding fatty acid and has an effect on connected pathways
HPLC-based quantification of lipid metabolites formed from fluorescent labeled compounds in living cells
Cells were cultivated for 24 h and harvested, and fatty aldehyde dehydrogenase activities were determined

Summary

Introduction

The lack of fatty aldehyde dehydrogenase function in Sjögren Larsson Syndrome (SLS) patient cells impairs the conversion of fatty aldehydes into their corresponding fatty acid and has an effect on connected pathways. We present a novel approach to examine fatty aldehyde metabolism in a time-dependent manner by measuring pyrene-labeled fatty aldehyde, fatty alcohol, fatty acid, and alkylglycerol in the culture medium of living cells using HPLC separation and fluorescence detection. In contrast, exclusively the corresponding fatty acid is formed. SLS patient cells did not display a hypersensitivity toward hexadecanal or hexadecanol, but 3-fold lower concentrations of the fatty alcohol than the corresponding fatty aldehyde were needed to induce toxicity in SLS patient and in control cells.—Keller, M. Studying fatty aldehyde metabolism in living cells with pyrene-labeled compounds.

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