Membrane contact sites: physical attachment between chloroplasts and endoplasmic reticulum revealed by optical manipulation.

Chloroplasts and their surrounding cell are highly interdependent. One example is lipid metabolism, where the cell depends on its chloroplasts to provide fatty acids for lipid synthesis in the endoplasmic reticulum (ER) and in turn, chloroplasts rely on import of lipid precursors from the ER. Despite its fundamental importance, the route for lipid trafficking into and out of chloroplasts remains unknown. Biochemical studies of plant membrane lipid metabolism have suggested the possibility of lipid transport at membrane contact sites (MCSs) between the ER and chloroplasts. With the aid of optical manipulation, we recently could present physical evidence for this association. Leaf protoplasts isolated from Arabidopsis thaliana expressing green fluorescent protein (GFP) in the ER lumen were observed by confocal microscopy. A laser scalpel was used to rupture the protoplasts. ER fragments associated with the released chloroplasts could be stretched out by optical tweezers but remained attached to the chloroplast surface, even when a stretching force of 400 pN was applied. We thus provided the first physical evidence for MCSs between two membranes and we propose for the ER-chloroplast pair, that such tight associations are involved in bidirectional lipid trafficking between the two compartments.