PUBLICATION
Developmental adcyap1b loss leads to hemorrhage, disrupted hemostasis, and a blood coagulation cascade in zebrafish
- Authors
- Ma, X., Guo, R., Xu, H., Ma, Y., Zhang, R., Liu, X., Zhang, J., Han, Y.
- ID
- ZDB-PUB-231218-13
- Date
- 2023
- Source
- Journal of thrombosis and haemostasis : JTH 22(4): 951-964 (Journal)
- Registered Authors
- Han, Ying, Ma, Yuanyuan, Zhang, Rui
- Keywords
- CRISPR/Cas9, adcyap1b, blood coagulation, hemostasis, zebrafish
- MeSH Terms
-
- Animals
- Anticoagulants/metabolism
- Blood Coagulation/genetics
- CRISPR-Associated Protein 9/metabolism
- Factor V/metabolism
- Hemorrhage
- Mammals/metabolism
- Morpholinos/genetics
- Morpholinos/metabolism
- Zebrafish*/genetics
- Zebrafish*/metabolism
- Zebrafish Proteins*/genetics
- Zebrafish Proteins*/metabolism
- PubMed
- 38104724 Full text @ J. Thromb. Haemost.
Citation
Ma, X., Guo, R., Xu, H., Ma, Y., Zhang, R., Liu, X., Zhang, J., Han, Y. (2023) Developmental adcyap1b loss leads to hemorrhage, disrupted hemostasis, and a blood coagulation cascade in zebrafish. Journal of thrombosis and haemostasis : JTH. 22(4):951-964.
Abstract
Background Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) is a neuropeptide with diverse roles in biological processes. Its involvement in the blood coagulation cascade is unclear.
Objectives This study unraveled adcyap1b's role in blood coagulation using CRISPR/Cas9 in zebrafish. Effects were validated via adcyap1b knockdown. Gene expression changes in adcyap1b mutants were explored, linking them to clotting disorders. An analysis of proca gene splicing illuminated its role in adcyap1b-related anticoagulation deficiencies.
Methods Zebrafish were genetically modified using CRISPR/Cas9 to induce adcyap1b knockout. Morpholino-mediated gene knockdown was employed for validation. Expression levels of coagulation factors, anticoagulant proteins, and fibrinolytic system genes were assessed in adcyap1b mutant zebrafish. Alternative splicing of proca gene was analyzed.
Results Adcyap1b mutant zebrafish exhibited severe hemorrhage, clotting disorders, and disrupted blood coagulation. Morpholino-mediated knockdown replicated observed phenotypes. Downregulation in transcripts related to coagulation factors V and IX, anticoagulation protein C, and plasminogen was observed. Abnormal alternative splicing of the proca gene was identified, providing a mechanistic explanation for anticoagulation system deficiencies.
Conclusions Adcyap1b plays a crucial role in maintaining zebrafish blood coagulation and hemostasis. Its influence extends to the regulation of procoagulant and anticoagulant pathways, with abnormal alternative splicing contributing to observed deficiencies. These findings unveil a novel aspect of adcyap1b function, offering potential insights into similar processes in mammalian systems.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping