ZFIN ID: ZDB-FIG-161202-31
Benz et al., 2016 - miR-19b Regulates Ventricular Action Potential Duration in Zebrafish. Scientific Reports   6:36033 Full text @ Sci. Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage: Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Long-pec to Day 5

Fig. 3

Loss of miR-19 leads to bradycardia.

(a) Lateral brightfield images of control-injected and MO19-injected 48 hpf zebrafish embryos. Note the cardiac edema and the blood congestion at the inflow tract developed in MO19 injected embryos. (b) Quantification of heart rate as heart beats per minute and ventricular fractional shortening as a measure for ventricular contractility in control (wt) and MO19 injected embryos. Morpholino-mediated knockdown of miR-19 in zebrafish leads to bradycardia and impaired ventricular contractility (±sd; n ≥ 14; p < 0.005). (c) qRT-PCR confirmed efficient knockdown of miR-19a-d. Notice the consistent reduction in miR-363 expression (±sd; n = 45 animals from 3 independent experiments). (d) Specific knockdown of miR-19a, miR-19c and miR-19d altered expression of neighboring miRNAs. However, knockdown of miR-19b did not interfere with expression of neighboring miRNAs (±sd; n = 45 animals from 3 independent experiments; p < 0.05). (e) 48 hpf zebrafish embryos injected with control- and MO19b-morpholino revealed that miR-19b deficiency is sufficient to mimic MO19-induced phenotype with characteristics of heart dysfunction. (f) miR-19b deficient embryos developed bradycardia with up to 36% ± 5.7% reduced heart frequency (± = sd; n ≥ 12; p < 0.005) (g,h) Relative changes in heart rate normalized to controls (dotted line) indicates that CRSIPR/Cas9 mediated knockout of miR-19b caused an identical and long-term reduction in heart rate as observed for MO19b (±sd; n = 10; p < 0.005).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
dre-mir-19a WT + MO1-dre-mir-19b standard conditions Long-pec whole organism RTPCR
WT + MO2-dre-mir-19b standard conditions Long-pec whole organism RTPCR
WT + MO2-mir19c standard conditions Long-pec whole organism RTPCR
WT + MO3-dre-mir-19a standard conditions Long-pec whole organism RTPCR
dre-mir-19b WT + MO1-dre-mir-19b standard conditions Long-pec whole organism RTPCR
WT + MO2-dre-mir-19b standard conditions Long-pec whole organism RTPCR
WT + MO2-mir19c standard conditions Long-pec whole organism RTPCR
WT + MO3-dre-mir-19a standard conditions Long-pec whole organism RTPCR
dre-mir-20a WT + MO2-mir19c standard conditions Long-pec whole organism RTPCR
WT + MO3-dre-mir-19a standard conditions Long-pec whole organism RTPCR
mir19c WT + MO1-dre-mir-19b standard conditions Long-pec whole organism RTPCR
WT + MO2-mir19c standard conditions Long-pec whole organism RTPCR
mir19d WT + MO1-dre-mir-19b standard conditions Long-pec whole organism RTPCR
WT + MO2-mir19c standard conditions Long-pec whole organism RTPCR
WT + MO2-mir19d standard conditions Long-pec whole organism RTPCR
mir25 WT + MO2-mir19c standard conditions Long-pec whole organism RTPCR
WT + MO2-mir19d standard conditions Long-pec whole organism RTPCR
mir363 WT + MO1-dre-mir-19b standard conditions Long-pec whole organism RTPCR
WT + MO2-mir19c standard conditions Long-pec whole organism RTPCR
WT + MO2-mir19d standard conditions Long-pec whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + CRISPR1-dre-mir-19 standard conditions Long-pec heart contraction decreased frequency, abnormal
Protruding-mouth heart contraction decreased frequency, abnormal
Day 5 heart contraction decreased frequency, abnormal
WT + MO1-dre-mir-19b standard conditions Long-pec cardiac ventricle cardiac muscle contraction decreased magnitude, abnormal
Long-pec eye decreased size, abnormal
Long-pec heart contraction decreased frequency, abnormal
Long-pec retina development in camera-type eye decreased process quality, abnormal
Long-pec whole organism dre-mir-19a expression decreased amount, abnormal
Long-pec whole organism dre-mir-19b expression decreased amount, abnormal
Long-pec whole organism mir19c expression decreased amount, abnormal
Long-pec whole organism mir19d expression decreased amount, abnormal
Long-pec whole organism mir363 expression decreased amount, abnormal
Protruding-mouth cardiac ventricle cardiac muscle contraction magnitude, normal
Protruding-mouth heart contraction decreased frequency, abnormal
WT + MO2-dre-mir-19b standard conditions Long-pec heart contraction decreased frequency, abnormal
Long-pec whole organism dre-mir-19b expression decreased amount, abnormal
Protruding-mouth heart contraction decreased frequency, abnormal
WT + MO2-mir19c standard conditions Long-pec whole organism dre-mir-19a expression decreased amount, abnormal
Long-pec whole organism dre-mir-19b expression decreased amount, abnormal
Long-pec whole organism dre-mir-20a expression increased amount, abnormal
Long-pec whole organism mir19c expression decreased amount, abnormal
Long-pec whole organism mir19d expression increased amount, abnormal
Long-pec whole organism mir25 expression increased amount, abnormal
Long-pec whole organism mir363 expression decreased amount, abnormal
WT + MO2-mir19d standard conditions Long-pec whole organism mir19d expression decreased amount, abnormal
Long-pec whole organism mir25 expression increased amount, abnormal
Long-pec whole organism mir363 expression increased amount, abnormal
WT + MO3-dre-mir-19a standard conditions Long-pec whole organism dre-mir-19a expression decreased amount, abnormal
Long-pec whole organism dre-mir-20a expression decreased amount, 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 @ Sci. Rep.