Topographical distribution of a membrane-inserted fluorescent phospholipid analogue during cell fusion

Abstract

Potential alterations in the transbilayer distribution of lipid molecules during cell-cell fusion were studied, using the fluorescent phospholipid analogue 1-acyl, 2-(N-4-nitrobenzo-2-oxa-1,3-diazole)-aminocaproyl phosphatidylcholine (C6-NBD-PC). The fluophore was inserted into the outer leaflet of the plasma membrane of Chinese hamster fibroblasts from an exogenous source and cell-cell fusion was induced either with Sendai virus or polyethylene glycol (PEG). After fusion, the cells were examined under a fluorescence microscope and the pool of tagged lipid molecules in the external monolayer was determined quantitatively. The results showed that in contrast to PEG-induced cell fusion, substantial redistribution of the lipid marker occurred when cell fusion was induced by Sendai virus and it was estimated that approx. 40% of exogenously supplied lipid was internalized. The possible mechanism causing lipid redistribution in the case of Sendai virus-induced cell fusion is discussed.