Metabolic modulation regulates cardiac wall morphogenesis in zebrafish
- Max Planck Institute for Heart and Lung Research, Germany
- Weizmann Institute of Science, Israel
Figures
Figure 1 with 3 supplements
Cardiomyocyte behavior during cardiac trabeculation.
(a) Schematic of the transplantation experiment. (b–e) 3D time-course images of chimeric hearts; magnified view (b’, c’, d’, e’) of area in white boxes and Y-Z plane images (b”, c”, d”, e”) along …
Figure 1—figure supplement 1
Cardiomyocytes change morphology and exhibit cell-cell junction rearrangements when entering the trabecular layer.
(a) Schematic of the transplantation experiment. (b) Circularity analysis of CMs in the compact and trabecular layers (n = 7–9 CMs from 3 to 5 ventricles). (c–c”) 3D images of chimeric hearts; Y-Z …
Figure 1—video 1
Z-stack images of 126 hpf CMs related to Figure 1b’.
Figure 1—video 2
Z-stack images of 175 hpf CMs related to Figure 1c’.
Figure 2 with 2 supplements
ERBB2 signaling activates glycolysis in cardiomyocytes.
(a) qPCR analysis of mRNA levels of glycolytic enzyme genes in control and Erbb2 overexpressing (OE) rat neonatal CMs (n = 3). Error bars, s.e.m. (b) Staining for PKM2, CTNI and DNA (DAPI) in …
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Figure 2—source data 1
Mass spectrometry data.
P7 WT and CAERBB2 OE mouse hearts were isolated and protein expression levels analyzed by mass spectrometry. All presented proteins are statistically significant at p<0.05.
- https://cdn.elifesciences.org/articles/50161/elife-50161-fig2-data1-v2.docx
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Figure 2—source data 2
Primer sequences for qPCR analysis.
Primer sequences used in Figure 2a and e.
- https://cdn.elifesciences.org/articles/50161/elife-50161-fig2-data2-v2.docx
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Figure 2—source data 3
Mean Ct values of qPCR analysis in Figure 2a and e.
- https://cdn.elifesciences.org/articles/50161/elife-50161-fig2-data3-v2.docx
Figure 2—figure supplement 1
NRG1/ERBB2 signaling activates glycolysis in CMs.
(a) ECAR analysis in control and NRG1 treated rat neonatal CMs; glycolytic capacity shown on the right (n = 7). (b) Confocal images (mid-sagittal sections) of 77 hpf zebrafish hearts treated with …
Figure 2—figure supplement 2
Uncropped images related to western blotting data.
Figure 3 with 1 supplement
Glycolysis regulates cardiac trabeculation.
(a) Confocal images (mid-sagittal sections) of 77 hpf hearts treated with DMSO, dichloroacetate (DCA) or Erbb2 inhibitor; magnified view of area in white boxes shown below; arrowheads point to CMs …
Figure 3—figure supplement 1
Cardiomyocyte proliferation does not appear to be affected by modulation of glycolysis.
(a) Confocal images (mid-sagittal sections) of 131 hpf Tg(myl7:EGFP-HRAS) alone or in combination with Tg(myl7:pdha1aSTA-P2A-tdTomato) and of 131 hpf Tg(myl7:EGFP-HRAS); pkma2-/-; pkmb-/- hearts; …
Figure 4 with 1 supplement
Loss of pkm2 impairs cardiac trabeculation.
(a) Schematic of the transplantation experiment. (b, b’) 3D and mid-sagittal section images of chimeric hearts using pkma2+/-; pkmb+/-; Tg(myl7:EGFP-HRAS) (b) and pkma2-/-; pkmb-/-; …
Figure 4—figure supplement 1
Glycolytic enzymes regulate cardiac trabeculation.
(a) Analysis of pkma and pkmb mRNA expression by in situ hybridization; magnified view shown in the top right corner; arrows point to the heart. (b) Expression levels of pkma and pkmb in 52 hpf and …
Author response image 1
Redox analysis.
70 hpf Tg(myl7:mitochondrial-Rogofp2Orp1) hearts were examined and 405 nm/488 nm ratios determined for redox state analysis. Relative values of 405 nm/488 nm in compact and trabecular cardiomyocytes …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (Danio rerio) | pkma2s717 | Stone et al., 2018 | ||
| Genetic reagent (Danio rerio) | pkmbs718 | Stone et al., 2018 | ||
| Gene (Danio rerio) | pdk3b | NM_001080688 | ||
| Gene (Danio rerio) | pdha1a | NM_213393 | ||
| Gene (Rattus norvegicus) | Pdk3 | NM_001106581 | ||
| Gene (Homo sapiens) | ERBB2 | NM_004448 | ||
| Antibody | anti-Cardiac Troponin I (Goat polyclonal) | Abcam | AB_880622 Cat# ab56357 | IF(1:500) |
| Antibody | anti-PKM2 (Rabbit monoclonal) | Cell Signaling | AB_1904096 Cat# D78A4 | IF(1:100) WB(1:1000) |
| Chemical compound, drug | Sodium dichloroacetate | Sigma Aldrich | 347795 | |
| Software, algorithm | Zen 2012 (Blue edition) | Carl Zeiss Microscopy | Version 1.1.2.0 | |
| Software, algorithm | Imaris x64 | Bitplane | Version 9.3.0 | |
| Other | DAPI stain | Sigma | D954 | (1 µg/mL) |
