Dynamics of asymmetric membranes and interleaflet coupling as intermediates in membrane fusion

Membrane fusion is a tool to increase the complexity of model membrane systems. Here, we use silica nanoparticles to fuse liquid-disordered DOPC giant unilamellar vesicles (GUVs) and liquid-ordered DPPC:cholesterol (7:3) GUVs. After fusion, GUVs display large membrane domains as confirmed by fluorescence confocal microscopy. Laurdan spectral imaging of the membrane phases in the fused GUVs shows differences compared with the initial vesicles indicating some lipid redistribution between phase domains as dictated by the tie lines of the phase diagram. Remarkably, using real-time confocal microscopy we were able to record the dynamics of formation of asymmetric membrane domains in hemifused GUVs and detected interleaflet coupling phenomena by which the DOPC-rich liquid-disordered domains in outer monolayer modulates the phase state of the DPPC:cholesterol inner membrane leaflet which transitions from liquid-ordered to liquid-disordered phase. We find that internal membrane stresses generated by membrane asymmetry enhance the efficiency of full fusion compared with our previous studies on symmetric vesicle fusion. Furthermore, under these conditions, the liquid-disordered monolayer dictates the bilayer phase state of asymmetric membrane domains in >90% of observed cases. By comparison to the findings of previous literature, we suggest that the monolayer phase that dominates the bilayer properties could be a mechanoresponsive signaling mechanism sensitive to the local membrane environment.