Related Experiment Video
Updated: Mar 23, 2026
Adult and Embryonic Skeletal Muscle Microexplant Culture and Isolation of Skeletal Muscle Stem Cells
Published on: September 21, 2010
Isolation and characterization of neural stem cells from dystrophic mdx mouse
Tiziana Annese1, Patrizia Corsi2, Simona Ruggieri1
1Department of Basic Medical Sciences, Neurosciences and Sensory Organs, Section of Human Anatomy and Histology, University of Bari Medical School, Bari, Italy.
Abstract:
The blood-brain barrier (BBB) is altered in mdx mouse, an animal model to study Duchenne muscular dystrophy (DMD). Our previous work demonstrated that perivascular glial endfeet control the selective exchanges between blood and neuropil as well as the BBB development and integrity; the alterations of dystrophin and dystrophin-associated protein complex (DAPs) in the glial cells of mdx mouse, parallel damages of the BBB and increase in vascular permeability. The aim of this study was to improve our knowledge about brain cellular components in the mdx mouse through the isolation, for the first time, of the adult neural stem cells (ANSCs). We characterized them by FACS, electron microscopy, confocal immunofluorescence microscopy, Real Time-PCR and western blotting, and we studied the expression of the DAPs aquaporin-4 (AQP4), potassium channel Kir4.1, α- and β-dystroglycan (αDG, βDG), α-syntrophin (αSyn), and short dystrophin isoform Dp71 proteins. The results showed that the mdx ANSCs expressed CD133 and Nestin receptor as the control ones, but showed a reduction in Notch receptor and altered cell proliferation with an increment in the apoptotic nuclei. Ultrastructurally, they appeared 50% size reduced compared to control ones, with a few cytoplasmic organelles. Moreover, the mdx ANSCs are devoid in full length dystrophin 427, and they expressed post-transcriptional reduction in the Dp71 in parallel with the ubiquitin proteasome activation, and decrement of DAPs proteins which appeared diffused in the cytoplasm and not polarized on the stem cells plasmamembrane, as prevalently observed in the controls. Overall, these results indicate that structural and molecular alterations affect the neural stem cells in the dystrophic brain, whose increased apoptosis and reduced Dp71 and DAPs proteins expression, together with loss in Dp427 dystrophin, could be responsible of the altered mdx glial maintenance and differentiation and consequent failure in the vessels barrier control occurring in the adult dystrophic brain.
Insights
Neural stem cells in Duchenne muscular dystrophy (DMD) mice show structural and molecular changes. These dystrophic brain cells exhibit increased apoptosis and altered protein expression, impacting blood-brain barrier integrity.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- The blood-brain barrier (BBB) is compromised in mdx mice, an animal model for Duchenne muscular dystrophy (DMD).
- Perivascular glial endfeet are crucial for BBB integrity, and their function is impaired by dystrophin and dystrophin-associated protein complex (DAPs) alterations in mdx mice.
Purpose of the Study:
- To investigate the characteristics of adult neural stem cells (ANSCs) in the mdx mouse model.
- To analyze the expression of key proteins, including dystrophin and DAPs, in mdx ANSCs.
Main Methods:
- Isolation and characterization of adult neural stem cells (ANSCs) from mdx and control mice.
- Utilized flow cytometry (FACS), electron microscopy, confocal immunofluorescence microscopy, Real Time-PCR, and western blotting.
- Assessed expression of aquaporin-4 (AQP4), Kir4.1, dystroglycans (αDG, βDG), α-syntrophin (αSyn), and Dp71.
Main Results:
- mdx ANSCs exhibited reduced Notch receptor expression, increased apoptosis, and smaller size with fewer organelles compared to controls.
- Full-length dystrophin (Dp427) was absent in mdx ANSCs, with reduced Dp71 expression and activated ubiquitin-proteasome system.
- DAPs were downregulated and mislocalized in mdx ANSCs, not polarized on the cell membrane as seen in controls.
Conclusions:
- Neural stem cells in the dystrophic brain exhibit significant structural and molecular abnormalities.
- Increased apoptosis and altered Dp71 and DAPs expression in mdx ANSCs contribute to glial dysfunction.
- These ANSC alterations likely lead to impaired blood-brain barrier control in adult dystrophic brains.
