FIGURE

Fig. 4

ID
ZDB-FIG-200728-38
Publication
Troutwine et al., 2020 - The Reissner Fiber Is Highly Dynamic In Vivo and Controls Morphogenesis of the Spine
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Fig. 4

Loss of the Reissner Fiber Is Associated with Scoliosis in Additional Independent Scoliosis Mutant Zebrafish Strains (A and A’) Inverted grayscale maximal Z projections of confocal images of Et(druk-GFPdut26/+); scospondin-GFPut24/+ demonstrating typical assembly of the Reissner fiber, floor plate, and terminal ampulla expression in these mutant knockin embryos at 5 dpf. Note that GFP expression is contributed both by the Et(druk-GFPdut26/+) transgene insertion and from the scospondin-GFPut24/+ allele. Scale bars, 25 μm in (A) and 5 μm in (A’). (B–D) Bright-field images of a wild-type juvenile scospondin-GFPut24/+ (B), Et(druk-GFPdut26/+) (C), and Et(druk-GFPdut26/+); scospondin-GFPut24/+ (D) displaying the onset of mild scoliosis. Scale bar, 1 mm; standard length in upper right corner. (B’–D’) Inverted grayscale maximal Z projections of confocal images of the same fish in (B), (C), and (D) to highlight the spinal cord are shown. At 19 dpf, in wild-type scospondin-GFPut24/+, we observe high expression of the Reissner fiber (100%; n = 14 size range 6.3–6.8 mm; B’); in scoliosis mutant Et(druk-GFPdut26/+); +/+, the Reissner fiber is not labeled but some spinal cord cells express GFP (n = 6 size range 6.0–6.5 mm); and in double mutant Et(druk-GFPdut26/+); scospondin-GFPut24/+, we observed curvature of the spinal canal and a complete loss of a Reissner fiber (94%; n = 16 size range 6.1–7.0 mm). Scale bars, 25 μm. (E and F) Inverted grayscale maximal Z projections from confocal imaging of AFRU-stained zebrafish embryos at 1 dpf (30 hpf). The Reissner fiber is observed in heterozygous kinesin family member 6 (kif6sko/+) (E) and homozygous kif6sko mutants (F) (8/8; for each genotype). Scale bars, 10 μm. (G–H”) Inverted grayscale maximal Z projections of confocal imaging of AFRU- and ZO-1-stained zebrafish at 3 dpf. ZO-1 localizes to tight junctions and shows the location of the central canal epithelium. Heterozygous kif6sko/+ animals have an intact Reissner fiber in the central canal (13/13; G–G”), whereas in homozygous kif6sko, the Reissner fiber is disassembled and Reissner material fills the space of the central canal (12/13; H–H”). Scale bars, 25 μm. (I and J) Merge of maximal Z projections of confocal imaging of AFRU- and ZO-1-stained zebrafish at 5 dpf. Heterozygous kif6sko/+ animals have an intact Reissner fiber in the central canal (I), but in homozygous kif6sko animals, diffuse Reissner material fills the central canal lumen (J). Scale bars, 10 μm.

Expression Data
Antibody:
Fish:
Anatomical Term:
Stage Range: Protruding-mouth to Day 5

Expression Detail
Antibody Labeling
Phenotype Data
Fish:
Observed In:
Stage Range: Prim-15 to Days 14-20

Phenotype Detail
Acknowledgments
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