Abstract
Understanding FA metabolism and lipid synthesis requires a lot of information about which FAs and lipids are formed within the cells. We focused on the use of deuterated substrates of 100 μM α-linolenic acid and linoleic acid to determine the relative amounts of their converted PUFAs and specific phospholipids that are incorporated into cell plasma membranes. Time-of-flight secondary ion mass spectrometry (TOF-SIMS) was used to image and analyze lipids in model cell membranes with and without FA treatment. Because of its high spatial resolution, TOF-SIMS can be used to simultaneously provide both chemical information and distribution of various molecules in the sample surface down to the subcellular scale. Data obtained from this analysis of isotopes in the cell samples were used to calculate the relative amounts of long-chain PUFAs and phospholipids from their precursors, α-linolenic acid and linoleic acid. Our results show that the FA treatments induced an increase in the amounts of α-linolenic acid and linoleic acid and their long-chain conversion products. Moreover, an enhanced level of phospholipid turnover of these FAs in lipids such as phosphatidylcholines, phosphatidylethanolamines, and phosphatidylinositols was also observed in the cell plasma membrane.
Highlights
Understanding FA metabolism and lipid synthesis requires a lot of information about which FAs and lipids are formed within the cells
The human body can synthesize all FAs it requires except for two, -linolenic acid (ALA; 18:3n-3), which belongs to the omega-3 (n-3) family, and linoleic acid (LA; 18:2n-6), which belongs to the omega-6 (n-6) family
The advantage of using deuterium-labeled FAs is the ability to determine the FA accumulation and incorporation into longer-chain FAs, as well as the relative amount of phospholipids that incorporate into the cell membrane
Summary
Understanding FA metabolism and lipid synthesis requires a lot of information about which FAs and lipids are formed within the cells. We focused on the use of deuterated substrates of 100 M -linolenic acid and linoleic acid to determine the relative amounts of their converted PUFAs and specific phospholipids that are incorporated into cell plasma membranes. Because of its high spatial resolution, TOF-SIMS can be used to simultaneously provide both chemical information and distribution of various molecules in the sample surface down to the subcellular scale Data obtained from this analysis of isotopes in the cell samples were used to calculate the relative amounts of long-chain PUFAs and phospholipids from their precursors, -linolenic acid and linoleic acid. Relative quantification of deuterated omega-3 and -6 FAs and their lipid turnover in PC12 cell membranes using TOF-SIMS.
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