Raw images from: Genetic inactivation of the β1 adrenergic receptor prevents Cerebral Cavernous Malformations in zebrafish
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Feb 24, 2025 version files 1.29 GB
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Abstract
Propranolol reduces experimental murine Cerebral Cavernous Malformations (CCMs) and prevents embryonic caudal venous plexus (CVP) lesions in zebrafish that follow mosaic inactivation of ccm2(Li et al., 2021). Because morpholino silencing of the β1 adrenergic receptor (adrb1) prevents the embryonic CVP lesion, we proposed that adrb1 plays a role in CCM pathogenesis. Here we report that adrb1-/- zebrafish exhibited 86% fewer CVP lesions and 87% reduction of CCM lesion volume relative to wild type brood mates at 2dpf and 8-10 weeks stage, respectively. Treatment with metoprolol, a β1 selective antagonist, yielded a similar reduction in CCM lesion volume. Adrb1-/- zebrafish embryos exhibited reduced heart rate and contractility and reduced CVP blood flow. Similarly, slowing the heart and eliminating the blood flow in CVP by administration of 2,3-BDM suppressed the CVP lesion. In sum, our findings provide genetic and pharmacological evidence that the therapeutic effect of propranolol on CCM is achieved through β1 receptor antagonism.
https://doi.org/10.5061/dryad.dz08kps7n
Description of the data and file structure
These images (CZI file) are from Zeiss LSM 880 Confocal with Airyscan, and can be viewed with ZEN or ImageJ.
Figure 1: Following ccm2 CRISPR injection, representative confocal images of 2 dpf Tg(fli1:EGFP) embryos illustrate that wild-type embryos (file “WT CCM2 CRISPR 2DPF-4_Airyscan Processing”) exhibit CVP dilation, whereas adrb1⁻/⁻ embryos (file “ADRB1 NULL CCM2 CRISPR 2DPF-2_Airyscan Processing”) are resistant to this defect.
Figure 4: Adrb1 signaling does not affect klf2a expression in ccm2 CRISPR embryos. Tg(klf2a:H2b-EGFP; kdrl:mCherry) embryos were injected, and nuclear EGFP signal in mCherry-labeled vascular endothelial cells was recorded using confocal microscopy. Representative images from each group are shown.
Control embryos (file “control mo 2dpf-5_Airyscan Processing”) injected with control MO alone served as the baseline. Ccm2 morphant embryos co-injected with adrb1 MO (file “ccm2 moadrb1 mo 2dpf-2_Airyscan Processing”) or control MO (file “ccm2 moctrl1 mo 2dpf-3_Airyscan Processing”) exhibited a significant increase in endothelial nuclear EGFP intensity compared to controls, with no significant difference between the two groups. All ccm2 CRISPR embryos were co-injected with tnnt MO, leading to an absence of blood flow. Compared to control embryos (file “control mo 2dpf-5_Airyscan Processing”), ccm2 CRISPR embryos co-injected with adrb1 MO (file “ccm2 CRISPR TNNT ADRB1 moS 2dpf-1_Airyscan Processing_Subset”) or control MO (file “klf2aEGFP flk1mcherry ccm2 crispr tnnt MO control moS 2dpf-2_Airyscan Processing_Subset”) displayed a mosaic increase in nuclear EGFP intensity in vascular endothelial cells, with no significant difference between the two groups.