Lowered GnT-I Activity Decreases Complex-Type N-Glycan Amounts and Results in an Aberrant Primary Motor Neuron Structure in the Spinal Cord

• Published in: Journal of developmental biology 2024-09, Vol.12 (3), p.21
• Main Authors: Hatchett, Cody J, Hall, M Kristen, Messer, Abel R, Schwalbe, Ruth A
• Format: Article
• Language: English
• Subjects: Cardiac muscle; Ethylenediaminetetraacetic acid; glycomics; Glycosylation; Hatching; Lectins; Lipids; Motor neurons; Motor skill; Muscles; Musculoskeletal system; Mutants; N-acetylglucosaminyltransferase-I (GnT-I); N-Acetylglucosinyldiphosphodolichol N-acetylglucosaminyltransferase; N-glycans; Nervous system; Neurons; oligomannose; Polysaccharides; Skeletal muscle; Skin; Skin tests; Spinal cord; Swim bladder; Swimming; Vertebrae; Zebrafish
• ISSN: 2221-3759; 2221-3759
• DOI: 10.3390/jdb12030021

The attachment of sugar to proteins and lipids is a basic modification needed for organismal survival, and perturbations in glycosylation cause severe developmental and neurological difficulties. Here, we investigated the neurological consequences of N-glycan populations in the spinal cord of Wt AB and mutant zebrafish. Mutant fish have reduced N-acetylglucosaminyltransferase-I (GnT-I) activity as remains intact. GnT-I converts oligomannose N-glycans to hybrid N-glycans, which is needed for complex N-glycan production. MALDI-TOF MS profiles identified N-glycans in the spinal cord for the first time and revealed reduced amounts of complex N-glycans in mutant fish, supporting a lesion in . Further lectin blotting showed that oligomannose N-glycans were more prevalent in the spinal cord, skeletal muscle, heart, swim bladder, skin, and testis in mutant fish relative to WT AB, supporting lowered GnT- I activity in a global manner. Developmental delays were noted in hatching and in the swim bladder. Microscopic images of caudal primary (CaP) motor neurons of the spinal cord transiently expressing EGFP in mutant fish were abnormal with significant reductions in collateral branches. Further motor coordination skills were impaired in mutant fish. We conclude that identifying the neurological consequences of aberrant N-glycan processing will enhance our understanding of the role of complex N-glycans in development and nervous system health.