title: Nrk2b-mediated NAD+ production regulates cell adhesion and is required for muscle morphogenesis in vivo: Nrk2b and NAD+ in muscle morphogenesis.
abstract: Cell-matrix adhesion complexes (CMACs) play fundamental roles during morphogenesis. Given the ubiquitous nature of CMACs and their roles in many cellular processes, one question is how specificity of CMAC function is modulated. The clearly defined cell behaviors that generate segmentally reiterated axial skeletal muscle during zebrafish development comprise an ideal system with which to investigate CMAC function during morphogenesis. We found that Nicotinamide riboside kinase 2b (Nrk2b) cell autonomously modulates the molecular composition of CMACs in vivo. Nrk2b is required for normal Laminin polymerization at the myotendinous junction (MTJ). In Nrk2b-deficient embryos, at MTJ loci where Laminin is not properly polymerized, muscle fibers elongate into adjacent myotomes and are abnormally long. In yeast and human cells, Nrk2 phosphorylates Nicotinamide Riboside and generates NAD+ through an alternative salvage pathway. Exogenous NAD+ treatment rescues MTJ development in Nrk2b-deficient embryos, but not in laminin mutant embryos. Both Nrk2b and Laminin are required for localization of Paxillin, but not beta-Dystroglycan, to CMACs at the MTJ. Overexpression of Paxillin in Nrk2b-deficient embryos is sufficient to rescue MTJ integrity. Taken together, these data show that Nrk2b plays a specific role in modulating subcellular localization of discrete CMAC components that in turn plays roles in musculoskeletal development. Furthermore, these data suggest that Nrk2b-mediated synthesis of NAD+ is functionally upstream of Laminin adhesion and Paxillin subcellular localization during MTJ development. These results indicate a previously unrecognized complexity to CMAC assembly in vivo and also elucidate a novel role for NAD+ during morphogenesis.
authors.affiliation: School of Biology and Ecology, University of Maine, Orono, ME 04469, USA.
authors.forename: Michelle F
authors.initials: MF
authors.lastname: Goody
authors.forename: Meghan W
authors.initials: MW
authors.lastname: Kelly
authors.forename: Kevin N
authors.initials: KN
authors.lastname: Lessard
authors.forename: Andre
authors.initials: A
authors.lastname: Khalil
authors.forename: Clarissa A
authors.initials: CA
authors.lastname: Henry
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D049030:Dystroglycans / Q000378:metabolism; D004625:Embryo, Nonmammalian; D007797:Laminin / Q000378:metabolism; D009024:Morphogenesis / Q000502:physiology*; D024510:Muscle Development / Q000502:physiology; D018482:Muscle, Skeletal / Q000254:growth & development* / Q000378:metabolism*; D009132:Muscles / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D051419:Paxillin / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor); D013710:Tendons / Q000378:metabolism / Q000502:physiology; D015027:Zebrafish / Q000378:metabolism
chemical_list: D007797:Laminin; D051419:Paxillin; D009243:NAD; D049030:Dystroglycans; D017853:Phosphotransferases (Alcohol Group Acceptor); C061783:nicotinamide riboside kinase