CHIR-124 Our data show that oxidative stress high levels of

Our data show that oxidative stress (high levels of ROS) suppresses the process of lysosomal exocytosis, which would be predicted to impair the expulsion of toxic compounds from cells. Therefore, we propose that ROS have a biphasic effect on the lysosomal clearance: consistent with their role as a second messenger moderate levels of ROS stimulate the lysosomal exocytosis, but pathological levels of ROS inhibit the lysosomal exocytosis. Therefore, the impact of oxidative stress pathology on CHIR-124 is two-pronged: beyond direct damage, it impairs the ability of the cell to limit and repair damage by extrusion of damaged materials. Indeed, an accumulation of damaged mitochondria and protein aggregates, which in turn can further amplify oxidative stress [24], is observed in multiple human diseases. An impact of oxidative stress on the cellular clearance pathway has been studied in many models; its specific effects are being disputed. An inhibitory effect of oxidative stress on the cellular clearance is also supported by data emerging from studies of ALS [72] and Alzheimer’s disease [66], [73]. Several candidate mechanisms for the observed effects can be discussed. Ca2+ release through TRPML1 has been implicated in lysosomal exocytosis [32]. Lysosomal exocytosis probably involves the stages of vesicle delivery to the plasma membrane and their fusion. TRPML1 has been implicated in vesicle positioning and motility [39] and membrane fusion [32]. Impairments in either of these processes would impact lysosomal exocytosis. Both processes appear to require Ca2+. Our previously published data [26] and the results presented here show that extracellular Ca2+ is necessary for the lysosomal exocytosis as well. These data suggest a contribution of the extracellular Ca2+ influx into cellular clearance and repair processes driven by lysosomal exocytosis. Oxidative stress was shown to inhibit the store-operated Ca2+ entry component Orai [74], [75], [76], and thus the inhibition of the store-operated Ca2+ entry by oxidative stress may be responsible for some of the loss of lysosomal exocytosis under oxidative stress. Since cell signaling involving G protein-coupled receptors drives the store-operated Ca2+ entry, this may serve as evidence connecting G protein-coupled receptor signaling and the cellular clearance via lysosomal exocytosis. We show that increasing extracellular Ca2+ stimulates lysosomal exocytosis to the extent that renders oxidative stress ineffective (Fig. 4). We think that the under these conditions, the increased Ca2+ influx triggers the processes normally actuated by TRPML1-dependent Ca2+ release, such as lysosomal positioning or fusion. Since the loss of lysosomal exocytosis was proposed to contribute to mucolipidosis type IV pathology [32], [38], it would be interesting to answer whether increased Ca2+ influx reverses some aspects of this pathology.
Conclusions
Approval statement
Conflict of interest statement
The Cortical Actin Network Regulates Exocytosis: From Micro- to Nanoscale Regulated exocytosis of secretory granules or vesicles is a fundamental cellular process involved in many functions, including neurotransmission, hormone secretion, cell migration, and thrombosis [1]. Successful secretion relies on the effective delivery of granules from inside the cell and the timely fusion of secretory vesicles with the plasma membrane. These processes involve the cortical actin network; a dense mesh of actin filaments that lies beneath the plasma membrane. This network has two seemingly opposing functions during regulated exocytosis: a negative role of the actin network providing a diffusion barrier that prevents the access of granules to secretory sites and traps secretory vesicles in cortical network 2, 3, 4, and the positive role works by driving exocytic vesicles to the fusion site, regulating the fusion pore, and providing the force to complete fusion 5, 6, 7. Such simultaneous opposing functions raise the question of how the actin cortex interacts with secretory granules and changes the environment to facilitate fusion and subsequent exocytosis 8, 9. The cortical actin network is key to recruiting secretory granules [10] in the immediate vicinity of the plasma membrane. It therefore constitutes a large area (microscopic) in which secretory vesicles are embedded in readiness to undergo myosin-II-dependent translocation to the plasma membrane in an activity-dependent manner 11, 12. Upon the arrival of these vesicles at hotspots of the plasma membrane 13, 14, a continuum of nanoscopic events allows docking and priming to occur. The environmental changes surrounding individual secretory vesicles are defined as nanoscopic, considering that the individual filaments display dynamic polymerization and depolymerization or reorganization around the granules, together with enrichment in molecules such as lipids and lipid-binding proteins. We therefore propose that the continuum of microscopic events that culminates in the nanoscale changes that occur around newly docked vesicles constitute a biological mesoscopic (see Glossary) effect that is specific to the continuum of interactions initiated by these vesicles at the cortical actin network. This in turn leads to the functionalization of hotspots (release sites) on the plasma membrane.