PUBLICATION
Mlck1a is expressed in zebrafish thrombocytes and an essential component for thrombus formation
- Authors
- Tournoij, E., Weber, G.J., Akkerman, J.W., de Groot, P.G., Zon, L.I., Moll, F.L., and Schulte-Merker, S.
- ID
- ZDB-PUB-091221-3
- Date
- 2010
- Source
- Journal of thrombosis and haemostasis : JTH 8(3): 588-595 (Journal)
- Registered Authors
- Schulte-Merker, Stefan, Zon, Leonard I.
- Keywords
- gene expression, mlck, Platelet, thrombosis, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Blood Platelets/enzymology*
- Cell Shape
- Disease Models, Animal
- Fibrinogen/metabolism
- Gene Expression Profiling/methods
- Gene Expression Regulation, Developmental
- Gene Expression Regulation, Enzymologic
- Gene Knockdown Techniques
- Myosin-Light-Chain Kinase/blood*
- Myosin-Light-Chain Kinase/genetics
- Oligonucleotide Array Sequence Analysis
- Platelet Adhesiveness
- Platelet Membrane Glycoprotein IIb/blood
- RNA, Messenger/blood
- Recombinant Fusion Proteins/blood
- Thrombosis/blood
- Thrombosis/enzymology*
- Thrombosis/genetics
- Zebrafish/blood*
- Zebrafish/embryology
- Zebrafish/genetics
- Zebrafish Proteins/blood*
- Zebrafish Proteins/genetics
- PubMed
- 20002541 Full text @ J. Thromb. Haemost.
Citation
Tournoij, E., Weber, G.J., Akkerman, J.W., de Groot, P.G., Zon, L.I., Moll, F.L., and Schulte-Merker, S. (2010) Mlck1a is expressed in zebrafish thrombocytes and an essential component for thrombus formation. Journal of thrombosis and haemostasis : JTH. 8(3):588-595.
Abstract
Summary Background: We have used the advantages of the zebrafish model system to demonstrate which of the vertebrate Myosin Light Chain Kinase (MLCK) genes is expressed in thrombocytes and important for thrombus formation. Methods and Results: Here we report that Mlck1a is an essential component for thrombus formation. Phylogenetic data revealed four zebrafish orthologous for three human MLCK genes. To investigate expression of the zebrafish mlck genes in thrombocytes we, compared GFP-tagged platelets with other cells by microarray analysis, and showed that mlck1a expression was 4.5 fold enriched in platelets. Furthermore, mlck1a mRNA and mRNA for the platelet-specific cd41 co-localized in thrombi. Expression of other mlck subtypes was lower in GFP-tagged platelets (mlck1b; 0.77 fold enriched) and absent in thrombi (mlck1b, -2, -3). To investigate the role of Mlck1a in thrombus formation, we knocked down mlck1a using two morpholinos. This resulted in impaired morphology changes of platelets adhering on fibrinogen. In a thrombosis model, in which thrombocytes adhere to the vessel wall damaged by laser irradiation, thrombus formation was slowed down in mlck1a deficient embryos. Conclusion: We conclude that Mlck1a is the subtype of MLCK that contributes to platelet shape change and thrombus formation.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping