Lateral phase separation of phospholipids as a basis for increased permeability of membranes towards fluorescein and other chemical species

Summary

Using mouse spleen cells, before and after treatment with glutaraldehyde or mild hyperthermia, we observe a strong correlation between permeability to fluorescein and susceptibility to staining with Nɛ-dansyl-l-lysine (irrespective of the cells' ability to exclude trypan blue). We observe the same correlation using liposomes prepared from phosphatidylcholine and varying amounts of cholesterol. We have recently introduced Nɛ-dansyl-l-lysine as a fluorescent membrane stain, or “probe,” whose uptake, we propose, is selective for phospholipid domains in membranes (G.M.K. Humphries & J.P. LovejoyBiophys. J. 42:307–310, 1983; G.M.K. Humphries & J.P. LovejoyJ. Membrane Biol. 77:115–122, 1984). The results presented here are consistent with the hypothesis that the presence or absence of phospholipid domains in membranes also modifies their permeability toward fluorescein, and suggests that permeability towards other chemical species may be similarly affected. On the basis of work using liposomes, we believe this to be the case for carboxy-fluorescein and for glucose.