PUBLICATION
Microarray analysis of prothrombin knockdown in zebrafish
- Authors
- Day, K.R., and Jagadeeswaran, P.
- ID
- ZDB-PUB-090518-21
- Date
- 2009
- Source
- Blood cells, molecules & diseases 43(2): 202-210 (Journal)
- Registered Authors
- Day, Kenneth, Jagadeeswaran, Pudur
- Keywords
- Zebrafish, Prothrombin, Morpholino, Embryonic development
- MeSH Terms
-
- Animals
- Down-Regulation
- Embryo, Nonmammalian/cytology
- Embryo, Nonmammalian/drug effects
- Embryo, Nonmammalian/metabolism
- Gene Expression Regulation, Developmental*
- Gene Knockdown Techniques
- Oligonucleotide Array Sequence Analysis
- Oligonucleotides, Antisense/genetics
- Prothrombin/genetics*
- Thrombin/antagonists & inhibitors
- Thrombin/metabolism*
- Up-Regulation
- Zebrafish/embryology*
- Zebrafish/genetics*
- PubMed
- 19442542 Full text @ Blood Cells Mol. Dis.
Citation
Day, K.R., and Jagadeeswaran, P. (2009) Microarray analysis of prothrombin knockdown in zebrafish. Blood cells, molecules & diseases. 43(2):202-210.
Abstract
The serine protease thrombin is generated from its precursor, prothrombin, in the coagulation cascade and plays a central role in fibrin deposition and platelet activation mediated through the protease activated receptors. Knockdown of prothrombin in the zebrafish was previously shown to recapitulate the phenotype observed in prothrombin knockout mice, such as an absence of blood pericardial edema, and hemorrhage. However, the role of thrombin during embryogenesis is not fully understood. To find genes affected by potential thrombin signaling in embryogenesis before blood circulation, microarray analysis was performed using total RNA prepared from antisense-injected, knockdown embryos versus mismatch-injected at 20 h post fertilization. A total of 63 upregulated and downregulated genes were identified with duplicate microarrays using dye reversal and a two-fold difference limitation. Real time RT-PCR for 10 selected genes identified by the microarray confirmed the expression changes in these genes. One particular gene, phlda3, was at least eleven fold upregulated, and in situ hybridization revealed expansion of phlda3 expression in the central nervous system, branchial arches, and head endoderm in knockdown embryos. The identification of these genes regulated by thrombin according to microarray analysis should provide a greater understanding of the effects of thrombin activity in the early vertebrate embryo.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping