Related Experiment Video
Updated: May 14, 2026
Cerebellar Regional Dissection for Molecular Analysis
Published on: December 5, 2020
Spatio-temporal analysis of molecular determinants of neuronal degeneration in the aging mouse cerebellum
Erik L de Graaf1, Wilbert P Vermeij, Monique C de Waard
1Biomolecular Mass Spectrometry and Proteomics Group, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Abstract:
The accumulation of cellular damage, including DNA damage, is hypothesized to contribute to aging-related neurodegenerative changes. DNA excision repair cross-complementing group 1 (Ercc1) knock-out mice represent an accepted model of neuronal aging, showing gradual neurodegenerative changes, including loss of synaptic contacts and cell body shrinkage. Here, we used the Purkinje cell-specific Ercc1 DNA-repair knock-out mouse model to study aging in the mouse cerebellum. We performed an in-depth quantitative proteomics analysis, using stable isotope dimethyl labeling, to decipher changes in protein expression between the early (8 weeks), intermediate (16 weeks), and late (26 weeks) stages of the phenotypically aging Ercc1 knock-out and healthy littermate control mice. The expression of over 5,200 proteins from the cerebellum was compared quantitatively, whereby 79 proteins (i.e. 1.5%) were found to be substantially regulated during aging. Nearly all of these molecular markers of the early aging onset belonged to a strongly interconnected network involved in excitatory synaptic signaling. Using immunohistological staining, we obtained temporal and spatial profiles of these markers confirming not only the proteomics data but in addition revealed how the change in protein expression correlates to synaptic changes in the cerebellum. In summary, this study provides a highly comprehensive spatial and temporal view of the dynamic changes in the cerebellum and Purkinje cell signaling in particular, indicating that synapse signaling is one of the first processes to be affected in this premature aging model, leading to neuron morphological changes, neuron degeneration, inflammation, and ultimately behavior disorders.
Insights
Cellular damage, particularly DNA damage, contributes to brain aging. This study reveals that impaired DNA repair in mice accelerates cerebellar aging, primarily affecting synaptic signaling and leading to neurodegeneration.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Cellular damage, including DNA damage, is a proposed driver of age-related neurodegeneration.
- DNA excision repair cross-complementing group 1 (Ercc1) deficient mice serve as a model for neuronal aging, exhibiting neurodegenerative traits.
- The cerebellum and its Purkinje cells are crucial for motor control and are affected in aging and neurodegenerative diseases.
Purpose of the Study:
- To investigate the molecular changes in the aging mouse cerebellum using a Purkinje cell-specific Ercc1 DNA-repair knock-out model.
- To identify early molecular markers of aging and neurodegeneration in the cerebellum.
- To correlate proteomic changes with synaptic and morphological alterations during premature aging.
Main Methods:
- Quantitative proteomics using stable isotope dimethyl labeling to compare protein expression across different aging stages (8, 16, 26 weeks).
- Utilized a Purkinje cell-specific Ercc1 knock-out mouse model and littermate controls.
- Immunohistological staining to validate proteomic findings and assess spatial-temporal protein distribution and synaptic changes.
Main Results:
- Over 5,200 cerebellar proteins were quantified, with 79 proteins (1.5%) significantly altered during aging.
- The majority of regulated proteins were part of an interconnected network involved in excitatory synaptic signaling.
- Proteomic data were confirmed by immunohistochemistry, demonstrating a correlation between protein expression changes and synaptic alterations.
Conclusions:
- Synaptic signaling is an early process affected in this premature aging model.
- Alterations in synaptic signaling precede and likely contribute to neuron morphological changes, degeneration, inflammation, and behavioral deficits.
- This study provides a comprehensive temporal and spatial view of cerebellar aging, highlighting the role of DNA repair deficiency in neurodegeneration.
