The Reissner Fiber Is Highly Dynamic In Vivo and Controls Morphogenesis of the Spine

Troutwine, Benjamin R; Gontarz, Paul; Konjikusic, Mia J.; Minowa, Ryoko; Monstad-Rios, Adrian T.; Sepich, Diane S.; Kwon, Ronald Y.; Solnica‐Krezel, Lilianna; Gray, Ryan S.

doi:10.1016/j.cub.2020.04.015

Cited by 70 publications

(154 citation statements)

References 40 publications

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“…SCO-spondin is the major protein component of the RF in zebrafish (Cantaut-Belarif et al, 2018). More comprehensive analysis of the non-complementing Group IA mutants, stl297 and stl300, found in this screen, demonstrated these are nonsynonymous mutations of the scospondin gene, causing disassembly of the RF during larval development leading to AIS-like spine defects (Troutwine et al, 2020). Given the similarity of Group IA phenotypes and the molecular implications for these mutations causing instability of the RF during larval development, it will be intriguing to determine whether the onset of whole- (Knafo and Wyart, 2018;Orts-Del'Immagine et al, 2020;Zhang et al, 2018).…”

Section: Discussionmentioning

confidence: 75%

“…We recently demonstrated that kif6 mutants have a defect in Reissner Fiber (RF) formation after 1 dpf (Troutwine et al, 2020), yet how Kif6 controls RF assembly is still under investigation. Kif6 is critical in zebrafish and mouse for the formation of ependymal cell cilia (Konjikusic et al, 2018), which are thought to contribute to 'near-wall' laminar flow of the cerebrospinal fluid in the ventricles and spinal cord (Spassky and Meunier, 2017).…”

Section: Discussionmentioning

confidence: 99%

“…3D), however the causative mutation remains undefined. Using WGS and WES we determined that both stl297 and stl300 were associated with non-synonymous mutations in the scospondin gene, which are described in greater detail in another manuscript (Troutwine et al, 2020). The remaining mutations have been cryopreserved and are being systematically introduced into our exome sequencing pipeline to facilitate mapping and identification of the causative mutations.…”

Section: Additional Group I Mutant Loci Map To Chromosomes 12 and 24mentioning

confidence: 99%

“…Given the dilated central canal in the adamts9 stl316/stl316 zebrafish mutants, we next wanted to assess whether central canal structures were defective. Recent studies demonstrate that disruption of a central canal resident structure called the Reissner fiber (RF) results in defects of spine morphogenesis (Troutwine et al, 2020). To determine whether the RF is defective in adamts9 stl316/stl316 mutants, we utilized the AFRU antiserum (Rodriguez et al, 1984) to stain zebrafish RF.…”

Section: Additional Group I Mutant Loci Map To Chromosomes 12 and 24mentioning

confidence: 99%

“…Partial complementation testing of the phenotypic group comprising 16 mutations, which displays the onset of pathology and morphology of scoliosis reminiscent of human AIS, defined at least 10 loci. Whole genome or exome sequencing of these scoliosis mutants, identified new mutations in the kif6 gene, which we previously implicated in zebrafish to model AIS (Buchan et al, 2014;Konjikusic et al, 2018), and two hypomorphic alleles of the scospondin gene, which encodes a critical protein component necessary for the development of the Reissner fiber (RF) and regulation of straightening of the body axis during embryonic development in zebrafish (Cantaut-Belarif et al, 2018;Troutwine et al, 2020). We also report several alleles of the a disintegrin and metalloproteinase with thrombospondin motifs 9 (adamts9) gene, which is associated with a unique scoliosis morphology beginning in the caudal portion of the spine.…”

Section: Introductionmentioning

confidence: 99%

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Postembryonic screen for mutations affecting spine development in zebrafish

Gray

González

Ackerman

et al. 2020

Preprint

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The spinal vertebral column gives structural support for the adult body plan, protects the spinal cord, and provides muscle attachment and stability, which allows the animal to move within its environment. The development and maturation of the spine and its physiology involve the integration of multiple musculoskeletal tissues including bone, cartilage, and fibrocartilaginous joints, as well as innervation and control by the nervous system. One of the most common disorders of the spine in human is adolescent idiopathic scoliosis (AIS), which is characterized by the onset of an abnormal lateral curvature of the spine of <10° around adolescence, in otherwise healthy children. The genetic basis of AIS is largely unknown. Systematic genome-wide mutagenesis screens for embryonic phenotypes in zebrafish have been instrumental in the understanding of early patterning of embryonic tissues necessary to build and pattern the embryonic spine. However, the mechanisms required for postembryonic maturation and homeostasis of the spine remain poorly understood. Here we report the results from a small-scale forward genetic screen for adult-viable recessive and dominant mutant zebrafish, displaying overt morphological abnormalities of the adult spine. Germline mutations induced with N-ethyl N-nitrosourea (ENU) were transmitted and screened for dominant phenotypes in 1,229 F1 animals, and subsequently bred to homozygosity in F3 families, from these, 314 haploid genomes were screened for recessive phenotypes. We cumulatively found 39 adult-viable (3 dominant and 36 recessive) mutations each leading to a defect in the morphogenesis of the spine. The largest phenotypic group displayed larval onset axial curvatures, leading to whole-body scoliosis without vertebral dysplasia in adult fish. Pairwise complementation testing within this phenotypic group revealed at least 16 independent mutant loci. Using massively-parallel whole genome or whole exome sequencing and meiotic mapping we defined the molecular identity of several loci for larval onset whole-body scoliosis in zebrafish. We identified a new mutation in the skolios/kinesin family member 6 (kif6) gene, causing neurodevelopmental and ependymal cilia defects in mouse and zebrafish. We also report several recessive alleles of the scospondin and a disintegrin and metalloproteinase with thrombospondin motifs 9 (adamts9) genes, which all display defects in spine morphogenesis. Many of the alleles characterized thus far are non-synonymous mutations in known essential scospondin and adamts9 genes. Our results provide evidence of monogenic traits that are critical for normal spine development in zebrafish, that may help to establish new candidate risk loci for spine disorders in humans.

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Section: Discussionmentioning

confidence: 75%

Section: Discussionmentioning

confidence: 99%

Section: Additional Group I Mutant Loci Map To Chromosomes 12 and 24mentioning

confidence: 99%

Section: Additional Group I Mutant Loci Map To Chromosomes 12 and 24mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Postembryonic screen for mutations affecting spine development in zebrafish

Gray

González

Ackerman

et al. 2020

Preprint

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Amphioxus neuroglia: Molecular characterization and evidence for early compartmentalization of the developing nerve cord

Bozzo

Lacalli

Obino

et al. 2021

Glia

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Glial cells play important roles in the development and homeostasis of metazoan nervous systems. However, while their involvement in the development and function in the central nervous system (CNS) of vertebrates is increasingly well understood, much less is known about invertebrate glia and the evolutionary history of glial cells more generally. An investigation into amphioxus glia is therefore timely, as this organism is the best living proxy for the last common ancestor of all chordates, and hence provides a window into the role of glial cell development and function at the transition of invertebrates and vertebrates. We report here our findings on amphioxus glia as characterized by molecular probes correlated with anatomical data at the transmission electron microscopy (TEM) level. The results show that amphioxus glial lineages express genes typical of vertebrate astroglia and radial glia, and that they segregate early in development, forming what appears to be a spatially separate cell proliferation zone positioned laterally, between the dorsal and ventral zones of neural cell proliferation. Our study provides strong evidence for the presence of vertebrate‐type glial cells in amphioxus, while highlighting the role played by segregated progenitor cell pools in CNS development. There are implications also for our understanding of glial cells in a broader evolutionary context, and insights into patterns of precursor cell deployment in the chordate nerve cord.

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Mutations in KIF7 implicated in idiopathic scoliosis in humans and axial curvatures in zebrafish

et al. 2021

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Idiopathic scoliosis (IS) is a spinal disorder affecting up to 3% of otherwise healthy children. IS has a strong familial genetic component and is believed to be genetically complex due to significant variability in phenotype and heritability. Previous studies identified putative loci and variants possibly contributing to IS susceptibility, including within extracellular matrix, cilia, and actin networks, but the genetic architecture and underlying mechanisms remain unresolved. Here, we used whole‐exome sequencing from three affected individuals in a multigenerational family with IS and identified 19 uncommon variants (minor allele frequency < 0.05). Genotyping of additional family members identified a candidate heterozygous variant (H1115Q, G>C, rs142032413) within the ciliary gene KIF7, a regulator within the hedgehog (Hh) signaling pathway. Resequencing of the second cohort of unrelated IS individuals and controls identified several severe mutations in KIF7 in affected individuals only. Subsequently, we generated a mutant zebrafish model of kif7 using CRISPR‐Cas9. kif7co63/co63 zebrafish displayed severe scoliosis, presenting in juveniles and progressing through adulthood. We observed no deformities in the brain, Reissner fiber, or central canal cilia in kif7co63/co63 embryos, although alterations were seen in Hh pathway gene expression. This study suggests defects in KIF7‐dependent Hh signaling, which may drive pathogenesis in a subset of individuals with IS.

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The Reissner Fiber Is Highly Dynamic In Vivo and Controls Morphogenesis of the Spine

Cited by 70 publications

References 40 publications

Postembryonic screen for mutations affecting spine development in zebrafish

Postembryonic screen for mutations affecting spine development in zebrafish

Amphioxus neuroglia: Molecular characterization and evidence for early compartmentalization of the developing nerve cord

Mutations in KIF7 implicated in idiopathic scoliosis in humans and axial curvatures in zebrafish

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