title: Loss of Mgat5a-mediatedN-glycosylation stimulates regeneration in zebrafish.
abstract: BACKGROUND
  We are using genetics to identify genes specifically involved in hearing regeneration. In a large-scale genetic screening, we identifiedmgat5a, a gene in theN-glycosylation biosynthesis pathway whose activity negatively impacts hair cell regeneration.
  
  
  METHODS
  We used a combination of mutant analysis in zebrafish and a hair cell regeneration assay to phenotype the loss of Mgat5a activity in zebrafish. We used pharmacological inhibition ofN-glycosylation by swansonine. We also used over-expression analysis by mRNA injections to demonstrate how changes inN-glycosylation can alter cell signaling.
  
  
  RESULTS
  We found thatmgat5awas expressed in multiple tissues during zebrafish embryo development, particularly enriched in neural tissues including the brain, retina, and lateral line neuromasts. Anmgat5ainsertional mutation and a CRISPR/Cas9-generated truncation mutation both caused an enhancement of hair cell regeneration which could be phenocopied by pharmacological inhibition with swansonine. In addition to hair cell regeneration, inhibition of theN-glycosylation pathway also enhanced the regeneration of lateral line axon and caudal fins. Further analysis showed thatN-glycosylation altered the responsiveness of TGF-beta signaling.
  
  
  CONCLUSIONS
  The findings from this study provide experimental evidence for the involvement ofN-glycosylation in tissue regeneration and cell signaling.
authors: Functional and Translation Genome Branch, National Human Genome Research Institute, 9000 Rockville Pike, Building 50, Room 5537, Bethesda, MD 20892 USA.
authors: Wuhong
authors: W
authors: Pei
authors: Functional and Translation Genome Branch, National Human Genome Research Institute, 9000 Rockville Pike, Building 50, Room 5537, Bethesda, MD 20892 USA.
authors: Sunny C
authors: SC
authors: Huang
authors: Functional and Translation Genome Branch, National Human Genome Research Institute, 9000 Rockville Pike, Building 50, Room 5537, Bethesda, MD 20892 USA.
authors: Lisha
authors: L
authors: Xu
authors: Functional and Translation Genome Branch, National Human Genome Research Institute, 9000 Rockville Pike, Building 50, Room 5537, Bethesda, MD 20892 USA.
authors: Kade
authors: K
authors: Pettie
authors: Center for Genome Regulation, Facultad de Ciencias, Universidad de Chile, Casilla 653, Santiago, Chile.
authors: María Laura
authors: ML
authors: Ceci
authors: Center for Genome Regulation, Facultad de Ciencias, Universidad de Chile, Casilla 653, Santiago, Chile.
authors: Mario
authors: M
authors: Sánchez
authors: Center for Genome Regulation, Facultad de Ciencias, Universidad de Chile, Casilla 653, Santiago, Chile.
authors: Miguel L
authors: ML
authors: Allende
authors: Functional and Translation Genome Branch, National Human Genome Research Institute, 9000 Rockville Pike, Building 50, Room 5537, Bethesda, MD 20892 USA.
authors: Shawn M
authors: SM
authors: Burgess
keywords: CRISPR/Cas9; N-glycosylation; Regeneration; Zebrafish; mgat5a