Fluorescent conjugates of brefeldin A selectively stain the endoplasmic reticulum and Golgi complex of living cells
| Autor: | Hee Chol Kang, Yupind Deng, R. P. Haugland, J. R. Bennink, J. W. Yewdell |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Boron Compounds
Histology Fluorescent Antibody Technique Golgi Apparatus Cyclopentanes Endoplasmic Reticulum Cell Line Mice chemistry.chemical_compound symbols.namesake L Cells Organelle Animals Lipid bilayer Chromatography High Pressure Liquid Fluorescent Dyes Protein Synthesis Inhibitors Brefeldin A Molecular Structure Staining and Labeling Endoplasmic reticulum Biological activity Golgi apparatus chemistry Cell culture symbols Biophysics Anatomy BODIPY |
| Zdroj: | Journal of Histochemistry & Cytochemistry. 43:907-915 |
| ISSN: | 1551-5044 0022-1554 |
| DOI: | 10.1177/43.9.7543914 |
| Popis: | The fungal metabolite brefeldin A (BFA) interferes with vesicular trafficking in most animal cells. To gain insight into the mechanism of BFA action, we esterified it to the fluorophore, boron dipyromethene difluoride (BODIPY). BODIPY-BEA localized predominantly in the endoplasmic reticulum (ER) and Golgi complex of viable cells and was extracted by detergent treatment, suggesting it interacts primarily with lipid bilayers. The localization of the conjugate is conferred by BFA, since free BODIPY or BODIPY esterified to cyclopentanol did not specifically localize to internal membranes. BODIPY-BFA exhibited a similar biological activity to BFA, but only when used at higher concentrations and after a delay. HPLC analysis revealed that over this period, cells converted BODIPY-BFA to species co-eluting with free BODIPY and BFA. Therefore, BODIPY-BFA is probably inactive until BFA is released by cellular esterases. The specific localization of BODIPY-BFA to the ER and Golgi complex suggests that BFA might exert its effects on vesicular trafficking by perturbing the lipid bilayer of its target organelles. Because BODIPY-BFA intensely stains the ER at concentrations that have no discernible effects on intracellular transport or other cellular functions, it should be useful for visualizing the ER in living cells. |
| Databáze: | OpenAIRE |
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