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Spot Variation Fluorescence Correlation Spectroscopy for Analysis of Molecular Diffusion at the Plasma Membrane of Living Cells
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Analysis of lipid-composition changes in plasma membrane microdomains
Hideo Ogiso1, Makoto Taniguchi2, Toshiro Okazaki3
1Department of Hematology/Immunology Kanazawa Medical University, 1-1 Daigaku, Uchinada, Ishikawa 920-0293, Japan.
Journal of Lipid Research
|June 28, 2015
Summary
This study reveals how plasma membrane microdomains adapt to lipid changes, finding glycosphingolipids can substitute for sphingomyelin. This research offers a new method for analyzing membrane lipids and their roles in cell signaling.
Area of Science:
- Cell Biology
- Biochemistry
- Lipidomics
Background:
- Plasma membrane microdomains, like lipid rafts and caveolae, are crucial for cell signaling.
- The precise lipid composition of these microdomains and how it changes during signaling are not fully understood.
Purpose of the Study:
- To investigate the lipid composition of plasma membrane microdomains.
- To understand how microdomain lipids change in response to stimuli, particularly in sphingomyelin-deficient cells.
Main Methods:
- A novel cationic colloidal silica bead method was developed to isolate and analyze plasma membrane lipids.
- Detergent-resistant fractions from silica bead-coated membranes were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Main Results:
- Glycosphingolipids were found to constitute microdomains in sphingomyelin-deficient cells, substituting for sphingomyelin (SM).
- Stimulation with C2-ceramide (Cer) in wild-type cells led to increased diacylglycerol and phosphatidic acid levels within microdomains.
- Serum stimulation induced similar lipid changes in microdomains of both SM-deficient and wild-type cells.
Conclusions:
- The developed method provides a practical approach for analyzing membrane microdomain lipids.
- This technique will aid future comprehensive studies on microdomain-associated cellular responses and lipid dynamics.
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