ZFIN ID: ZDB-FIG-170815-9
Garcia et al., 2017 - Sheath Cell Invasion and Trans-differentiation Repair Mechanical Damage Caused by Loss of Caveolae in the Zebrafish Notochord. Current biology : CB   27(13):1982-1989.e3 Full text @ Curr. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: Protruding-mouth to Days 7-13
PHENOTYPE:
Fish:
Observed In:
Stage Range: Protruding-mouth to Days 7-13

Fig. 4

Invading Sheath Cells Differentiate into Vacuolated Cells

(A and B) Projections of confocal stacks of 72 hpf WT and cavin1b−/− larvae expressing rcn3:GFP-rab32a and col9a2:mcherry. Brackets mark areas of vacuolated cell collapse and sheath invasion; arrows point to new vacuoles in sheath cells.

(C and D) Following invasion, sheath cell vacuoles (arrows) enlarge. Intact primary vacuolated cells are traced with dashed lines. The asterisks mark an intact vacuolated cell.

(E) Confocal images of live sheath cells obtained from 48 hpf embryos expressing col9a2:mcherry and rcn3:GFP-rab32a. Cells were treated with suramin and AR-C 118925XX (AR-C) or ATPγS/UTPγS or left untreated for 2 hr. Arrows point to newly formed vacuoles.

(F) Quantitation using one-way ANOVA followed by multiple comparisons using Tukey’s test, ∗∗∗p < 0.001; n.s., not significant; n = 3 experiments.

(G–O) LSM imaging of cavin1b mutants expressing col8a1a:GFPCaax and col9a2:mcherry.

(G–I) 3.6-mm fish. Arrows point to vacuolated sheath cells; asterisks mark a primary vacuolated cell.

(J–L) 4.5-mm cavin1b−/− fish showing new vacuolated cells that retained col9a2:mcherry expression (arrows) next to a primary vacuolated cell (asterisks).

(M–O) 4.3-mm cavin1b−/− fish with remnants of primary vacuolated cells (arrow), a primary vacuolated cell (asterisk), and newly differentiated vacuolated cells (blue arrows).

Scale bars, 50 μm. Error bars are SD. See also Figures S3 and S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
GFP cavin1bbns110/bns110; pd1150Tg; pd1152Tg (EKW) standard conditions Day 4 notochord inner cell IFL
Day 4 Not Detected notochord outer sheath cell IFL
Days 7-13 notochord inner cell IFL
Days 7-13 notochord outer sheath cell IFL
cavin1bbns110/bns110; pd1150Tg; pd1153Tg (EKW) standard conditions Protruding-mouth notochord outer sheath cell vacuole IFL
pd1150Tg; pd1153Tg (EKW) standard conditions Protruding-mouth notochord outer sheath cell vacuole IFL
mCherry cavin1bbns110/bns110; pd1150Tg; pd1152Tg (EKW) standard conditions Day 4 notochord outer sheath cell IFL
Days 7-13 notochord outer sheath cell IFL
cavin1bbns110/bns110; pd1150Tg; pd1153Tg (EKW) standard conditions Protruding-mouth notochord outer sheath cell IFL
pd1150Tg; pd1153Tg (EKW) standard conditions Protruding-mouth notochord outer sheath cell IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
cavin1bbns110/bns110; pd1150Tg; pd1152Tg (EKW) standard conditions Day 4 notochord inner cell metaplastic, abnormal
Day 4 notochord outer sheath cell displaced to notochord central region, abnormal
Day 4 notochord outer sheath cell metaplastic, abnormal
Days 7-13 notochord inner cell metaplastic, abnormal
Days 7-13 notochord outer sheath cell displaced to notochord central region, abnormal
Days 7-13 notochord outer sheath cell GFP expression increased amount, abnormal
Days 7-13 notochord outer sheath cell metaplastic, abnormal
cavin1bbns110/bns110; pd1150Tg; pd1153Tg (EKW) standard conditions Protruding-mouth notochord inner cell collapsed, abnormal
Protruding-mouth notochord outer sheath cell displaced to notochord central region, abnormal
Protruding-mouth notochord outer sheath cell metaplastic, abnormal
Protruding-mouth notochord outer sheath cell vacuole GFP expression increased amount, abnormal
Protruding-mouth notochord outer sheath cell vacuole increased size, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Curr. Biol.