Related Experiment Video
Updated: Jun 20, 2025
Experimental Approaches to Study Mitochondrial Localization and Function of a Nuclear Cell Cycle Kinase, Cdk1
Published on: February 25, 2016
Mutant NOTCH3ECD Triggers Defects in Mitochondrial Function and Mitophagy in CADASIL Cell Models
Wan Wang1, Zhenping Gong2, Yadan Wang3
1Department of Neurology, Zhengzhou University People's Hospital, Henan Provincial People's Hospital, Zhengzhou, Henan, China.
Insights
NOTCH3ECD deposition in Cerebral Autosomal-Dominant Arteriopathy with Subcortical Infarction and Leukoencephalopathy (CADASIL) impairs mitochondrial function and mitophagy. This study reveals a common pathology across different NOTCH3 mutations, impacting brain small vessel disease.
Area of Science:
- Neuroscience
- Cell Biology
- Genetics
Background:
- Cerebral Autosomal-Dominant Arteriopathy with Subcortical Infarction and Leukoencephalopathy (CADASIL) is a genetic small-vessel brain disease.
- NOTCH3 extracellular domain (NOTCH3ECD) deposition is the primary pathology in CADASIL.
- The specific impact of NOTCH3ECD on mitochondrial function in CADASIL remains unclear.
Purpose of the Study:
- To investigate the role of mitochondrial dysfunction in CADASIL.
- To determine how NOTCH3ECD deposition affects mitochondrial structure and function.
Main Methods:
- Established human embryonic kidney-293T cell models with NOTCH3ECD alterations.
- Assessed mitochondrial function using flow cytometry and structure via transmission electron microscopy.
- Evaluated mitophagy using western blotting and immunofluorescence.
Main Results:
- NOTCH3ECD deposition altered mitochondrial morphology and function.
- NOTCH3ECD protein levels correlated with mitochondrial quality and directly bound to mitochondria.
- Autophagy and mitophagy were induced but impaired, leading to abnormal mitochondrial accumulation.
Conclusions:
- NOTCH3ECD deposition presents a common pathological feature across different NOTCH3 mutations in CADASIL.
- This study provides novel insights into NOTCH3ECD's role in mitochondrial dysfunction and mitophagy.
- Findings highlight potential therapeutic targets for CADASIL by addressing mitochondrial health.
Background:
Cerebral autosomal-dominant arteriopathy with subcortical infarction and leukoencephalopathy (CADASIL) is an inherited small-vessel disease that affects the white matter of the brain. Recent studies have confirmed that the deposition of NOTCH3ECD is the main pathological basis of CADASIL; however, whether different mutations present the same pathological characteristics remains to be further studied. Some studies have found that mitochondrial dysfunction is related to CADASIL; however, the specific effects of NOTCH3ECD on mitochondrial remain to be determined.
Objective:
We aimed to explore the role of mitochondrial dysfunction in CADASIL.
Methods:
We established transgenic human embryonic kidney-293T cell models (involving alterations in cysteine and non-cysteine residues) via lentiviral transfection. Mitochondrial function and structure were assessed using flow cytometry and transmission electron microscopy, respectively. Mitophagy was assessed using western blotting and immunofluorescence.
Results:
We demonstrated that NOTCH3ECD deposition affects mitochondrial morphology and function, and that its protein levels are significantly correlated with mitochondrial quality and can directly bind to mitochondria. Moreover, NOTCH3ECD deposition promoted the induction of autophagy and mitophagy. However, these processes were impaired, leading to abnormal mitochondrial accumulation.
Conclusions:
This study revealed a common pathological feature of NOTCH3ECD deposition caused by different NOTCH3 mutations and provided new insights into the role of NOTCH3ECD in mitochondrial dysfunction and mitophagy.
Related Concept Videos
03:14Notch Signaling Pathway
The Notch gene came into the limelight in 1914 after the discovery that its mutation in Drosophila melanogaster leads to a serrated (or "notched") wing margin phenotype. It was not...
01:12Role Of Notch Signalling In Intestinal Stem Cell Renewal
Direct cell-to-cell contact is needed for the activation of Notch signaling. The signal is initiated when a notch ligand binds to a receptor on an adjacent cell, also...
01:46Electron Transport Chain: Complex I and II
ROS generation is regulated and maintained at moderate levels necessary...