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Endothelial cell plasma membrane obtained by chemically induced vesiculation.

N I Moldovan, A N Radu, N Simionescu

    Experimental Cell Research
    |June 1, 1987
    PubMed
    Summary

    Researchers developed a chemical method to isolate right-side out luminal plasmalemma vesicles from endothelial cells (EC). This technique provides pure EC plasma membrane for studying its composition and function in vascular beds.

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    Area of Science:

    • Biochemistry
    • Cell Biology
    • Vascular Biology

    Background:

    • Endothelial cells (EC) form the inner lining of blood vessels.
    • Understanding the composition and function of the EC plasma membrane is crucial for vascular health.

    Purpose of the Study:

    • To develop a method for isolating right-side out luminal plasmalemma-derived vesicles from EC.
    • To characterize these vesicles for purity and fidelity to the native EC plasma membrane.

    Main Methods:

    • Chemical vesiculation using low concentrations of formaldehyde and dithiothreitol.
    • Microscopy techniques including phase-contrast, transmission electron microscopy (TEM), and scanning electron microscopy (SEM).
    • Biochemical and cytochemical analyses, including freeze-fracture, Ruthenium Red staining, lectin agglutination, and 125I-labeling.

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    Main Results:

    • Vesicles were successfully isolated and exhibited the characteristic trilaminar membrane structure without organelles.
    • Vesicle membranes contained intramembrane particles and cholesterol, and their surfaces showed characteristic binding patterns with Ruthenium Red and ferritin conjugates.
    • Vesicles retained EC glycoconjugates and expressed 5'-nucleotidase activity, with similar external polypeptide profiles to intact EC.

    Conclusions:

    • Chemically induced vesiculation is a suitable method for obtaining pure endothelial cell (EC) luminal plasmalemma.
    • These isolated vesicles can be used to study the composition and functions of the EC plasma membrane in various vascular beds.