Monitoring phospholipid dynamics in vivo with a fluorescent dye octadecyl rhodamine B

Nov 3, 2025Cell structure and function

Tracking phospholipid changes in living organisms using a glowing dye

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Abstract

R18 is internalized in a phosphatidylethanolamine (PE)-dependent manner in vivo.

Internalization of R18 in Saccharomyces cerevisiae is blocked in PE-deficient mutants.
Supplementing with ethanolamine restores R18 internalization, indicating a strict need for PE.
R18 delivered to vacuoles through autophagy is not retained but is exported in a manner dependent on Pep4 and Atg15.
Exported R18 is redirected to endosomes after prolonged autophagy.
R18 may indicate PE dynamics and contribute to lipid transport between vacuoles and endosomes.

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Phospholipids are major components of biological membranes. They play an essential role in intracellular signaling and organelle dynamics; however, the availability of suitable lipid-specific probes is limited, which has hindered studies on their spatial distribution and functional dynamics in living cells. Previously, we demonstrated that octadecyl rhodamine B chloride (R18) is transported to the endoplasmic reticulum via nonvesicular membrane transport. In this study, we showed that R18 is internalized in a phosphatidylethanolamine (PE)-dependent manner in vivo. The internalization of R18 in Saccharomyces cerevisiae is blocked in PE-deficient mutants, but restored by ethanolamine supplementation, which suggests strict PE dependence. Moreover, R18 delivered to vacuoles through autophagy was not terminally retained, but underwent Pep4- and Atg15-dependent export from the vacuoles. The exported R18 was then redirected to endosomes following prolonged autophagy. These results suggest that R18 may serve as an indicator of PE dynamics and vacuole-endosome lipid transport, which contributes to lipid homeostasis inside vacuoles.Key words: autophagy, in vivo lipid dynamics, octadecyl rhodamine B (R18), phospholipase, phospholipid, vacuole, yeast.

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Monitoring phospholipid dynamics in vivo with a fluorescent dye octadecyl rhodamine B

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