PUBLICATION
Loss of calpain3b in Zebrafish, a Model of Limb-Girdle Muscular Dystrophy, Increases Susceptibility to Muscle Defects Due to Elevated Muscle Activity
- Authors
- Prykhozhij, S.V., Caceres, L., Ban, K., Cordeiro-Santanach, A., Nagaraju, K., Hoffman, E.P., Berman, J.N.
- ID
- ZDB-PUB-230226-53
- Date
- 2023
- Source
- Genes 14(2): (Journal)
- Registered Authors
- Ban, Kevin, Berman, Jason, Caceres, Lucia, Prykhozhij, Sergey
- Keywords
- Duchenne muscular dystrophy, birefringence, calpain 3, cholinesterase inhibitor, disease model, limb-girdle muscle dystrophy, methylcellulose, muscle, zebrafish
- MeSH Terms
-
- Animals
- Muscle, Skeletal/metabolism
- Muscular Dystrophies, Limb-Girdle*/genetics
- Muscular Dystrophy, Duchenne*/pathology
- Zebrafish/genetics
- PubMed
- 36833417 Full text @ Genes (Basel)
Citation
Prykhozhij, S.V., Caceres, L., Ban, K., Cordeiro-Santanach, A., Nagaraju, K., Hoffman, E.P., Berman, J.N. (2023) Loss of calpain3b in Zebrafish, a Model of Limb-Girdle Muscular Dystrophy, Increases Susceptibility to Muscle Defects Due to Elevated Muscle Activity. Genes. 14(2):.
Abstract
Limb-Girdle Muscular Dystrophy Type R1 (LGMDR1; formerly LGMD2A), characterized by progressive hip and shoulder muscle weakness, is caused by mutations in CAPN3. In zebrafish, capn3b mediates Def-dependent degradation of p53 in the liver and intestines. We show that capn3b is expressed in the muscle. To model LGMDR1 in zebrafish, we generated three deletion mutants in capn3b and a positive-control dmd mutant (Duchenne muscular dystrophy). Two partial deletion mutants showed transcript-level reduction, whereas the RNA-less mutant lacked capn3b mRNA. All capn3b homozygous mutants were developmentally-normal adult-viable animals. Mutants in dmd were homozygous-lethal. Bathing wild-type and capn3b mutants in 0.8% methylcellulose (MC) for 3 days beginning 2 days post-fertilization resulted in significantly pronounced (20-30%) birefringence-detectable muscle abnormalities in capn3b mutant embryos. Evans Blue staining for sarcolemma integrity loss was strongly positive in dmd homozygotes, negative in wild-type embryos, and negative in MC-treated capn3b mutants, suggesting membrane instability is not a primary muscle pathology determinant. Increased birefringence-detected muscle abnormalities in capn3b mutants compared to wild-type animals were observed following induced hypertonia by exposure to cholinesterase inhibitor, azinphos-methyl, reinforcing the MC results. These mutant fish represent a novel tractable model for studying the mechanisms underlying muscle repair and remodeling, and as a preclinical tool for whole-animal therapeutics and behavioral screening in LGMDR1.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping