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ZFIN ID: ZDB-FIG-110622-76
Fish et al., 2011 - A Slit/miR-218/Robo regulatory loop is required during heart tube formation in zebrafish. Development (Cambridge, England)   138(7):1409-1419 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: 20-25 somites
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: 20-25 somites

Fig. 3

Slit/Robo signaling regulates heart tube formation. (A) Early heart field migration defects in zebrafish morphants: slit2, miR-218, robo1 and robo4. Shown is Tg(kdrl:ras-mCherry);Tg(myl7:GFP) expression at 20 somites. Dorsal views, anterior to the top. slit2 morphants have disrupted endocardial migration, including loss of sheet-like migration and endocardial cells have multiple filopodial protrusions (arrows). miR-218 and robo1 morphants have delayed migration of the endocardium and myocardium. The endocardial cells of robo1 morphants display a rounded phenotype (arrow). Migration of the heart fields is not delayed in robo4 morphants, but the morphology of the endocardial and myocardial cells is altered compared with controls. (B) Migration velocities of endocardial [Tg(kdrl:ras-mCherry) expression] and myocardial [Tg(myl7:GFP) expression] cells were quantified from time-lapse images. The number of cells tracked is shown above each bar. slit2 morphants have enhanced endocardial migration rates, whereas miR-218 and robo1 have reduced endocardial and myocardial migration rates. *, P<0.05 compared with control. (C) Directionality of migration was assessed by quantifying the ratio of the net distance traveled to the total distance traveled. A decrease in the ratio indicates an increase in the randomness of migration. slit2 and robo4 morphants have defects in directional migration. *, P<0.05 compared with control. Data in B and C are mean + s.e.m. (D) In situ analysis of robo1 expression at the 20-somite stage reveals an increase in expression in miR-218 morphants. Transverse sections, dorsal to the top. ec, endocardium; m, myocardium. Scale bars: 50 μm in A and D.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP s843Tg control 20-25 somites myocardium IFL
s843Tg + MO1-mir218 standard conditions 20-25 somites myocardium IFL
s896Tg; twu26Tg control 20-25 somites myocardium IFL
s896Tg; twu26Tg + MO1-mir218 standard conditions 20-25 somites myocardium IFL
s896Tg; twu26Tg + MO1-robo1 standard conditions 20-25 somites myocardium IFL
s896Tg; twu26Tg + MO2-slit2 standard conditions 20-25 somites myocardium IFL
s896Tg; twu26Tg + MO3-robo4 standard conditions 20-25 somites myocardium IFL
mCherry s896Tg; twu26Tg control 20-25 somites endocardium IFL
s896Tg; twu26Tg + MO1-mir218 standard conditions 20-25 somites endocardium IFL
s896Tg; twu26Tg + MO1-robo1 standard conditions 20-25 somites endocardium IFL
s896Tg; twu26Tg + MO2-slit2 standard conditions 20-25 somites endocardium IFL
s896Tg; twu26Tg + MO3-robo4 standard conditions 20-25 somites endocardium IFL
robo1 s843Tg control 20-25 somites endocardium ISH
20-25 somites myocardium ISH
s843Tg + MO1-mir218 standard conditions 20-25 somites endocardium ISH
20-25 somites myocardium ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
s896Tg; twu26Tg + MO1-mir218 standard conditions 20-25 somites cardiac muscle tissue morphogenesis decreased process quality, abnormal
20-25 somites cardioblast migration to the midline involved in heart field formation decreased process quality, abnormal
20-25 somites endocardium mislocalised, abnormal
20-25 somites endocardium morphogenesis decreased process quality, abnormal
20-25 somites myocardium mislocalised, abnormal
s896Tg; twu26Tg + MO1-robo1 standard conditions 20-25 somites cardiac muscle tissue morphogenesis decreased process quality, abnormal
20-25 somites cardioblast migration to the midline involved in heart field formation decreased process quality, abnormal
20-25 somites endocardium mislocalised, abnormal
20-25 somites endocardium cell malformed, abnormal
20-25 somites endocardium morphogenesis decreased process quality, abnormal
20-25 somites myocardium mislocalised, abnormal
s896Tg; twu26Tg + MO2-slit2 standard conditions 20-25 somites endocardial progenitor cell migration to the midline involved in heart field formation decreased process quality, abnormal
s896Tg; twu26Tg + MO2-slit2 standard conditions 20-25 somites endocardium mislocalised, abnormal
s896Tg; twu26Tg + MO2-slit2 standard conditions 20-25 somites endocardium cell malformed, abnormal
s896Tg; twu26Tg + MO2-slit2 standard conditions 20-25 somites endocardium formation decreased process quality, abnormal
s896Tg; twu26Tg + MO3-robo4 standard conditions 20-25 somites cardioblast migration decreased process quality, abnormal
20-25 somites endocardium cell malformed, abnormal
20-25 somites myocardium cell malformed, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development