Induction of Vesicle Formation by Exposing Apple Tissue to Vacuum Impregnation

This paper demonstrates that vacuum impregnation of mature apple tissue in the presence of different sugars results in the formation of membrane vesicles inside the cells. Vesiculation is regarded to be a metabolic consequence of the impregnation process. Vesiculation is shown when the endocytic marker FM4-64 was impregnated into the apple tissue together with the sugar solutions. Vesicles were formed at the plasma membrane already 30 min after impregnation and remained inside the cells for at least 24 h, a metabolic process that was inhibited in the presence of chloroquine, a specific endosomic inhibitor. This phenomenon was not dependent on the osmotic strength when sucrose was used for impregnation. However, the vesiculation drastically dropped when a hypertonic trehalose solution was impregnated. We suggest that the impregnated sugars may not totally remain in the extracellular space between the cells, as normally believed, but at least a fraction might be incorporated into the cells.