PUBLICATION

NAD+ improves neuromuscular development in a zebrafish model of FKRP-associated dystroglycanopathy

Authors
Bailey, E.C., Alrowaished, S.S., Kilroy, E.A., Crooks, E.S., Drinkert, D.M., Karunasiri, C.M., Belanger, J.J., Khalil, A., Kelley, J.B., Henry, C.A.
ID
ZDB-PUB-190809-1
Date
2019
Source
Skeletal muscle   9: 21 (Journal)
Registered Authors
Henry, Clarissa A.
Keywords
Dystroglycanopathy, FKRP, NAD+, Neuromuscular junction, Zebrafish
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Disease Models, Animal
  • Dystroglycans/deficiency*
  • Dystroglycans/genetics*
  • Glycosylation
  • Glycosyltransferases/genetics*
  • Glycosyltransferases/metabolism*
  • Humans
  • Muscle Development/genetics
  • Muscle Development/physiology
  • Muscular Dystrophy, Animal/genetics*
  • Muscular Dystrophy, Animal/metabolism*
  • Muscular Dystrophy, Animal/pathology
  • Mutation
  • NAD/administration & dosage
  • NAD/metabolism*
  • Neuromuscular Junction/genetics
  • Neuromuscular Junction/growth & development
  • Neuromuscular Junction/metabolism
  • Paxillin/genetics
  • Paxillin/metabolism
  • Up-Regulation
  • Zebrafish
  • Zebrafish Proteins/deficiency*
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism*
PubMed
31391079 Full text @ Skelet Muscle
Abstract
Secondary dystroglycanopathies are muscular dystrophies that result from mutations in genes that participate in Dystroglycan glycosylation. Glycosylation of Dystroglycan is essential for muscle fibers to adhere to the muscle extracellular matrix (myomatrix). Although the myomatrix is disrupted in a number of secondary dystroglycanopathies, it is unknown whether improving the myomatrix is beneficial for these conditions. We previously determined that either NAD+ supplementation or overexpression of Paxillin are sufficient to improve muscle structure and the myomatrix in a zebrafish model of primary dystroglycanopathy. Here, we investigate how these modulations affect neuromuscular phenotypes in zebrafish fukutin-related protein (fkrp) morphants modeling FKRP-associated secondary dystroglycanopathy.
We found that NAD+ supplementation prior to muscle development improved muscle structure, myotendinous junction structure, and muscle function in fkrp morphants. However, Paxillin overexpression did not improve any of these parameters in fkrp morphants. As movement also requires neuromuscular junction formation, we examined early neuromuscular junction development in fkrp morphants. The length of neuromuscular junctions was disrupted in fkrp morphants. NAD+ supplementation prior to neuromuscular junction development improved length. We investigated NMJ formation in dystroglycan (dag1) morphants and found that although NMJ morphology is disrupted in dag1 morphants, NAD+ is not sufficient to improve NMJ morphology in dag1 morphants. Ubiquitous overexpression of Fkrp rescued the fkrp morphant phenotype but muscle-specific overexpression only improved myotendinous junction structure.
These data indicate that Fkrp plays an early and essential role in muscle, myotendinous junction, and neuromuscular junction development. These data also indicate that, at least in the zebrafish model, FKRP-associated dystroglycanopathy does not exactly phenocopy DG-deficiency. Paxillin overexpression improves muscle structure in dag1 morphants but not fkrp morphants. In contrast, NAD+ supplementation improves NMJ morphology in fkrp morphants but not dag1 morphants. Finally, these data show that muscle-specific expression of Fkrp is insufficient to rescue muscle development and homeostasis.
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