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Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis
Published on: August 7, 2018
RIPK3 activation promotes DAXX-dependent neuronal necroptosis after intracerebral hemorrhage in mice
Qingqing Bai1,2, Shuoyang Wang1, Dongmei Rao1
1Department of Neurology, First Affiliated Hospital of Wannan Medical College, Yijishan Hospital, Wuhu, Anhui, China.
Background:
Necroptosis induced by receptor-interacting protein kinase 3 (RIPK3) is engaged in intracerebral hemorrhage (ICH) pathology. In this study, we explored the impact of RIPK3 activation on neuronal necroptosis and the mechanism of the death domain-associated protein (DAXX)-mediated nuclear necroptosis pathway after ICH.
Methods:
Potential molecules linked to the progression of ICH were discovered using RNA sequencing. The level of DAXX was assessed by quantitative real-time PCR, ELISA, and western blotting. DAXX localization was determined by immunofluorescence and immunoprecipitation assays. The RIPK3 inhibitor GSK872 and DAXX knockdown with shRNA-DAXX were used to examine the nuclear necroptosis pathway associated with ICH. Neurobehavioral deficit assessments were performed.
Results:
DAXX was increased in patients and mice after ICH. In an ICH mouse model, shRNA-DAXX reduced brain water content and alleviated neurologic impairments. GSK872 administration reduced the expression of DAXX. shRNA-DAXX inhibited the expression of p-MLKL. Immunofluorescence and immunoprecipitation assays showed that RIPK3 and AIF translocated into the nucleus and then bound with nuclear DAXX.
Conclusions:
RIPK3 revitalization promoted neuronal necroptosis in ICH mice, partially through the DAXX signaling pathway. RIPK3 and AIF interacted with nuclear DAXX to aggravate ICH injury.
Insights
Receptor-interacting protein kinase 3 (RIPK3) activation drives neuronal necroptosis in intracerebral hemorrhage (ICH) via the DAXX pathway. Targeting RIPK3 and DAXX may offer new therapeutic strategies for ICH.
Area of Science:
- Neuroscience
- Cellular Biology
- Pathology
Background:
- Necroptosis, a programmed cell death pathway regulated by RIPK3, plays a role in intracerebral hemorrhage (ICH).
- The specific mechanisms of RIPK3-mediated neuronal death and the involvement of the DAXX-nuclear necroptosis pathway in ICH remain unclear.
Purpose of the Study:
- To investigate the role of RIPK3 activation in neuronal necroptosis following ICH.
- To elucidate the mechanism of the DAXX-mediated nuclear necroptosis pathway in the context of ICH.
Main Methods:
- RNA sequencing to identify ICH-related molecules.
- Quantitative PCR, ELISA, and western blotting to assess DAXX levels.
- Immunofluorescence and immunoprecipitation assays for DAXX localization.
- Utilized RIPK3 inhibitor GSK872 and DAXX knockdown (shRNA-DAXX) in an ICH mouse model.
- Performed neurobehavioral assessments.
Main Results:
- DAXX expression was elevated in patients and mice post-ICH.
- shRNA-DAXX treatment reduced brain water content and improved neurological function in an ICH mouse model.
- GSK872 decreased DAXX expression, and shRNA-DAXX inhibited p-MLKL.
- RIPK3 and AIF translocated to the nucleus, binding with nuclear DAXX.
Conclusions:
- RIPK3 activation promotes neuronal necroptosis in ICH, partly via the DAXX signaling pathway.
- RIPK3 and AIF interaction with nuclear DAXX exacerbates ICH-induced brain injury.
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