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Updated: Oct 18, 2025
Force Measurement During Contraction to Assess Muscle Function in Zebrafish Larvae
Published on: July 23, 2013
A CRISPR/Cas9 zebrafish lamin A/C mutant model of muscular laminopathy
Hannah A Nicolas1, Khang Hua1, Hailey Quigley1
1Department of Biology, Faculty of Science, University of Ottawa, Ottawa, Ontario, Canada.
Background:
Lamin A/C gene (LMNA) mutations frequently cause cardiac and/or skeletal muscle diseases called striated muscle laminopathies. We created a zebrafish muscular laminopathy model using CRISPR/Cas9 technology to target the zebrafish lmna gene.
Results:
Heterozygous and homozygous lmna mutants present skeletal muscle damage at 1 day post-fertilization (dpf), and mobility impairment at 4 to 7 dpf. Cardiac structure and function analyses between 1 and 7 dpf show mild and transient defects in the lmna mutants compared to wild type (WT). Quantitative RT-PCR analysis of genes implicated in striated muscle laminopathies show a decrease in jun and nfκb2 expression in 7 dpf homozygous lmna mutants compared to WT. Homozygous lmna mutants have a 1.26-fold protein increase in activated Erk 1/2, kinases associated with striated muscle laminopathies, compared to WT at 7 dpf. Activated Protein Kinase C alpha (Pkc α), a kinase that interacts with lamin A/C and Erk 1/2, is also upregulated in 7 dpf homozygous lmna mutants compared to WT.
Conclusions:
This study presents an animal model of skeletal muscle laminopathy where heterozygous and homozygous lmna mutants exhibit prominent skeletal muscle abnormalities during the first week of development. Furthermore, this is the first animal model that potentially implicates Pkc α in muscular laminopathies.
Insights
We developed a zebrafish model for muscular laminopathies caused by Lamin A/C gene (LMNA) mutations. This model shows skeletal muscle damage and mobility issues, potentially implicating Pkc α in the disease.
Area of Science:
- Genetics and Molecular Biology
- Developmental Biology
- Cardiovascular Biology
Background:
- Mutations in the Lamin A/C gene (LMNA) are a common cause of striated muscle laminopathies, affecting cardiac and skeletal muscles.
- A zebrafish model was engineered using CRISPR/Cas9 to study muscular laminopathies.
Purpose of the Study:
- To establish a zebrafish model for muscular laminopathies.
- To investigate the effects of lmna gene disruption on skeletal and cardiac muscle development and function.
- To explore potential molecular pathways involved in lmna-related muscle diseases.
Main Methods:
- CRISPR/Cas9 gene editing was used to target the zebrafish lmna gene, creating heterozygous and homozygous mutants.
- Skeletal muscle integrity and mobility were assessed from 1 to 7 days post-fertilization (dpf).
- Cardiac structure and function were analyzed, and gene expression (jun, nfκb2) and protein levels (Erk 1/2, Pkc α) were quantified using RT-PCR and Western blotting.
Main Results:
- lmna mutants displayed skeletal muscle damage at 1 dpf and impaired mobility from 4 to 7 dpf.
- Mild, transient cardiac defects were observed in lmna mutants compared to wild type (WT) zebrafish.
- Homozygous lmna mutants showed decreased jun and nfκb2 expression, increased activated Erk 1/2, and upregulated Protein Kinase C alpha (Pkc α) compared to WT at 7 dpf.
Conclusions:
- The developed zebrafish model exhibits significant skeletal muscle abnormalities in early development, consistent with skeletal muscle laminopathy.
- This model is the first to suggest a potential role for Pkc α in muscular laminopathies.
