Mixed lineage kinase domain-like protein induces RGC-5 necroptosis following elevated hydrostatic pressure.

Mixed lineage kinase domain-like protein induces RGC-5 necroptosis following elevated hydrostatic pressure. Acta Biochim Biophys Sin (Shanghai). 2017 Oct 01;49(10):879-889 Authors: Liao L, Shang L, Li N, Wang S, Wang M, Huang Y, Chen D, Huang J, Xiong K Abstract Receptor-interacting protein 3 (RIP3) is an essential component of the necroptosis signaling pathway. Phosphorylation of its downstream target, mixed lineage kinase domain-like protein (MLKL), has been proposed to induce necroptosis by initiating Ca2+ influx. Our previous studies have shown that RGC-5 retinal ganglion cells undergo RIP3-mediated necroptosis following elevated hydrostatic pressure (EHP). However, the molecular mechanism underlying necroptosis induction downstream of RIP3 is still not well understood. Here, we investigated the effects of MLKL during EHP-induced necroptosis, and primarily explored the relationship between MLKL and Ca2+ influx. Immunofluorescence staining showed that the expression of MLKL was increased 12 h after EHP. Western blot analysis demonstrated that the phosphorylated and unphosphorylated forms of both RIP3 and MLKL were up-regulated 12 h after EHP, while inhibition of RIP3 by GSK'872 decreased the expression of phosphorylated MLKL at the same stage. Propidium iodide staining, lactate dehydrogenase release assays, flow cytometry, and electron microscopy revealed the increased necrosis of RGC-5 cells 12 h after EHP, which coincided with e...
Source: Acta Biochimica et Biophysica Sinica - Category: Biochemistry Authors: Tags: Acta Biochim Biophys Sin (Shanghai) Source Type: research