Developmental expression of vitamin K-dependent gamma-carboxylase activity in zebrafish embryos: Effect of warfarin

Ravikumar Hanumanthaiah, Bharath Thankavel, Kenneth Day, Michael Gregory, Pudur Jagadeeswaran

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Vitamin K-dependent gamma-carboxylation is an essential posttranslational modification required for the functional activity of coagulation proteins such as factors VII, IX, X, and prothrombin. Warfarin, an inhibitor of vitamin K-dependent gamma-carboxylation, was used in earlier work on adult zebrafish to provide evidence for the presence of vitamin K-dependent carboxylase in zebrafish. Here we demonstrate the presence of vitamin K-dependent carboxylase activity in zebrafish by directly assaying the microsomal fraction prepared from adult, unfertilized eggs, and embryos from different developmental stages. Gamma-carboxylase activity was detected both before and after fertilization of embryos and the activity levels remained relatively constant from 6 h postfertilization (hpf) through other advanced stages of development. The expression of activity in the early embryos (0-6 hpf) may be due to the presence of maternal protein since the activity was detected even in the unfertilized eggs. Gamma-carboxylase activity in the eggs as well as early embryos suggested that vitamin K-dependent carboxylase is important throughout development. The detection of vitamin K-dependent carboxylase mRNA by RT-PCR and inhibitor studies using warfarin confirmed these activity results. Further, these studies provide a basis for selecting warfarin-resistant zebrafish mutants in order to find genes regulating gamma-carboxylase activity including the yet unidentified vitamin K-epoxide reductase.

Original languageEnglish (US)
Pages (from-to)992-999
Number of pages8
JournalBlood Cells, Molecules, and Diseases
Volume27
Issue number6
DOIs
StatePublished - 2001

Fingerprint

Zebrafish
Warfarin
Embryonic Structures
Vitamin K
Ovum
Vitamin K Epoxide Reductases
Factor VII
Factor IX
Prothrombin
Post Translational Protein Processing
Fertilization
Eggs
Proteins
Mothers
Polymerase Chain Reaction
Messenger RNA
glutamyl carboxylase
Genes

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Hematology

Cite this

Developmental expression of vitamin K-dependent gamma-carboxylase activity in zebrafish embryos : Effect of warfarin. / Hanumanthaiah, Ravikumar; Thankavel, Bharath; Day, Kenneth; Gregory, Michael; Jagadeeswaran, Pudur.

In: Blood Cells, Molecules, and Diseases, Vol. 27, No. 6, 2001, p. 992-999.

Research output: Contribution to journalArticle

Hanumanthaiah, Ravikumar ; Thankavel, Bharath ; Day, Kenneth ; Gregory, Michael ; Jagadeeswaran, Pudur. / Developmental expression of vitamin K-dependent gamma-carboxylase activity in zebrafish embryos : Effect of warfarin. In: Blood Cells, Molecules, and Diseases. 2001 ; Vol. 27, No. 6. pp. 992-999.
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