PUBLICATION
asb5a/asb5b Double Knockout Affects Zebrafish Cardiac Contractile Function
- Authors
- Cai, W., Wang, Y., Luo, Y., Gao, L., Zhang, J., Jiang, Z., Fan, X., Li, F., Xie, Y., Wu, X., Li, Y., Yuan, W.
- ID
- ZDB-PUB-231126-7
- Date
- 2023
- Source
- International Journal of Molecular Sciences 24(22): (Journal)
- Registered Authors
- Wu, Xiushan
- Keywords
- CRISPR/Cas9, RNA-seq, asb5a, asb5b, hub genes, morpholino, semi-automatic heartbeat analysis
- MeSH Terms
-
- Protein Isoforms/metabolism
- Zebrafish*/metabolism
- Muscle Contraction
- Zebrafish Proteins*/genetics
- Zebrafish Proteins*/metabolism
- Animals
- Organogenesis
- PubMed
- 38003559 Full text @ Int. J. Mol. Sci.
Citation
Cai, W., Wang, Y., Luo, Y., Gao, L., Zhang, J., Jiang, Z., Fan, X., Li, F., Xie, Y., Wu, X., Li, Y., Yuan, W. (2023) asb5a/asb5b Double Knockout Affects Zebrafish Cardiac Contractile Function. International Journal of Molecular Sciences. 24(22):.
Abstract
Ankyrin repeat and suppression-of-cytokine-signaling box (Asb) proteins, a subset of ubiquitin ligase E3, include Asb5 with six ankyrin-repeat domains. Zebrafish harbor two asb5 gene isoforms, asb5a and asb5b. Currently, the effects of asb5 gene inactivation on zebrafish embryonic development and heart function are unknown. Using CRISPR/Cas9, we generated asb5a-knockout zebrafish, revealing no abnormal phenotypes at 48 h post-fertilization (hpf). In situ hybridization showed similar asb5a and asb5b expression patterns, indicating the functional redundancy of these isoforms. Morpholino interference was used to target asb5b in wild-type and asb5a-knockout zebrafish. Knocking down asb5b in the wild-type had no phenotypic impact, but simultaneous asb5b knockdown in asb5a-knockout homozygotes led to severe pericardial cavity enlargement and atrial dilation. RNA-seq and cluster analyses identified significantly enriched cardiac muscle contraction genes in the double-knockout at 48 hpf. Moreover, semi-automatic heartbeat analysis demonstrated significant changes in various heart function indicators. STRING database/Cytoscape analyses confirmed that 11 cardiac-contraction-related hub genes exhibited disrupted expression, with three modules containing these genes potentially regulating cardiac contractile function through calcium ion channels. This study reveals functional redundancy in asb5a and asb5b, with simultaneous knockout significantly impacting zebrafish early heart development and contraction, providing key insights into asb5's mechanism.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping