Autophagy is an evolutionarily conserved lysosomal degradation pathway, yet the underlying mechanisms remain poorly understood. In previous studies, we showed the Nicotinic acid adenine dinucleotide phosphate (NAADP) as a potent Ca2+ mobilizing messenger in astrocytes, since NAADP releases Ca2+ from the endo-lysosomal system and this effect was mediated by the two-pore channels (TPCs), especially TPC type 2. Some independent studies have been shown that the specificity of NAADP-mediated Ca2+ release is inhibited by Ned-19. But, whether and how TPCs and NAADP act in astrocytes is unclear and the functions on autophagy pathway have yet to be defined. To better understand how TPCs functions are regulated, a proteomic screening by Mass Spectrometric of TPC1 and TPC2 interacting proteins will be performed. Because TPCs might be NAADP-sensitive Ca2+-permeable channel in lysosomes, will be examined the NAADP-AM on interacting proteins regulation in TPC1 or 2-overexpressing cells. Furthermore, the levels of interacting proteins autophagy-related also will be studied under NAADP or under autophagic inducers effects through western blotting and immunoprecipitation experiments. Taken together, our results further will demonstrate that the activation of NAADP/TPCs signaling might compromise the differently the autophagy progression, suggesting the potential physiological of NAADP/TPCs in central nervous system.
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