Cholesterol stabilizes recombinant exocytic fusion pores by altering membrane bending rigidity

No abstract

Face masks are a primary preventive measure against airborne pathogens. Thus, they have become one of the keys to controlling the spread of the COVID-19 virus. Common examples, including N95 masks, surgical masks, and face coverings, are passive devices that minimize the spread of suspended pathogens by inserting an aerosol-filtering barrier between the user’s nasal and oral cavities and the environment. However, the filtering process does not adapt to changing pathogen levels or other environmental factors, which reduces its effectiveness in real-world scenarios. This paper addresses the limitations of passive masks by proposing ADAPT, a smart IoT-enabled “active mask”. This wearable device contains a real-time closed-loop control system that senses airborne particles of different sizes near the mask by using an on-board particulate matter (PM) sensor. It then intelligently mitigates the threat by using mist spray, generated by a piezoelectric actuator, to load nearby aerosol particles such that they rapidly fall to the ground. The system is controlled by an on-board micro-controller unit that collects sensor data, analyzes it, and activates the mist generator as necessary. A custom smartphone application enables the user to remotely control the device and also receive real-time alerts related to recharging, refilling, and/or decontamination of the mask before reuse. Experimental results on a working prototype confirm that aerosol clouds rapidly fall to the ground when the mask is activated, thus significantly reducing PM counts near the user. Also, usage of the mask significantly increases local relative humidity levels.

Cholesterol is essential for neuronal activity and function. Cholesterol depletion in the plasma membrane impairs synaptic transmission. However, the molecular mechanisms by which cholesterol deficiency leads to defects in vesicle fusion remain poorly understood. Here we show that cholesterol is required for Ca2+-dependent fusion using the in-vitro reconstitution of vesicle fusion, atomic force microscopy (AFM), and amperometry to monitor exocytosis in chromaffin cells. Purified native vesicles were crucial for the complete reconstitution of physiological Ca2+-dependent fusion, whereas vesicle-mimicking liposomes failed to reproduce the cholesterol effect. Intriguingly, cholesterol had no effect on membrane insertion of synaptotagmin-1, a Ca2+ sensor for ultrafast fusion. Cholesterol stabilizes local membrane bending induced by synaptotagmin-1 insertion, thereby lowering the energy barrier for Ca2+-dependent fusion to occur. Our data provide evidence that cholesterol depletion abolishes Ca2+-dependent vesicle fusion by disrupting synaptotagmin-1-induced membrane bending, and suggests that cholesterol is a master regulator for Ca2+-dependent fusion.