ZFIN ID: ZDB-FIG-180621-81
Müller-Deile et al., 2018 - Overexpression of preeclampsia induced microRNA-26a-5p leads to proteinuria in zebrafish. Scientific Reports   8:3621 Full text @ Sci. Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Day 4
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Day 4

Fig. 4

Vegf-Aa knockdown by vegf-Aa-MO or miR-26a-5p mimic cause impairments in intersegmental tail vessel sprouting. (A) Western blot for Vegf-Aa and normalization with Gapdh of whole zebrafish larvae lysate (10 µg of a pool of 10 fish). Zebrafish were injected with either a recombinant zebrafish Vegf-Aa protein (Vegf-Aa protein), a control morpholino (CTRL-MO), a vegf-Aa morpholino (vegf-Aa-MO), a control miR mimic (miR-CTRL) or a miR-26a-5p mimic (miR-26a) at the one to four cell stages. Zebrafish were harvested at 96 hpf. Quantification of western blot bands are shown below the blot. n = 12 fish per group, total n = 48. Full-length blots are presented in supplementary Fig. 3. (B) Phenotype pictures of zebrafish show blood pooling in tail region (arrow heads) in zebrafish that were injected with a vegf-Aa morpholino (vegf-Aa-MO) or a miR-26a-5p mimic (miR-26a-5p) were as control injected fish (CTRL-MO and miR-CTRL) appeared normal. Scale bar: 4 mM. (C) Pictures of tail vessel region of Tg(l-fabp:DBP-eGFP/flk-mcherry) zebrafish. These transgenic fish express a red fluorescent endothelium and green fluorescent plasma protein. Zebrafish were injected with either a control morpholino (CTRL-MO), a vegf-Aa morpholino (vegf-Aa-MO) a miR-26a-5p mimic (miR-26a-5p), a control miR mimic (miR-CTRL) alone (−) or together with a recombinant zebrafish Vegf-Aa protein (Vegf-Aa) (+) at the one to four cell stages. Vessel pictures were taken at 96 hpf. Arrow heads illustrate defects in sprouting of intersegmental vessel in vegf-Aa-MO and miR-26a-5p mimic injected fish. DLAV: dorsal lateral anatomic vessel. Scale bar = 500 µM. (D) Representative tail vessel pictures of Tg(l-fabp:DBP-eGFP/flk-mcherry) zebrafish with normal intersegmental tail vessel sprouting (left) and quantification of intersegmental vessels sprouting in zebrafish that were injected with miR mimics and morpholinos as indicated (right). n = 12 per group, total n = 96.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP lri500Tg; zf2134Tg + MO10-vegfaa standard conditions Day 4 post-vent vasculature blood plasma IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-vegf WB WT + MO10-vegfaa standard conditions Day 4 whole organism
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO10-vegfaa standard conditions Day 4 caudal fin blood accumulation caudal fin, abnormal
Day 4 post-vent vasculature blood circulation decreased occurrence, abnormal
Day 4 whole organism ab1-vegf labeling decreased amount, abnormal
lri500Tg; zf2134Tg + MO10-vegfaa standard conditions Day 4 intersegmental vessel sprouting angiogenesis decreased occurrence, abnormal
Day 4 post-vent vasculature blood plasma EGFP expression decreased amount, abnormal
Day 4 post-vent vasculature intersegmental vessel morphology, abnormal
Day 4 post-vent vasculature morphology, abnormal
Acknowledgments:
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