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Identification of menaquinone-4 (vitamin K2) target genes in bovine endometrial epithelial cells in vitro
Hanako Bai1, Manabu Kawahara1, Masashi Takahashi2
1Laboratory of Animal Breeding and Reproduction, Research Faculty of Agriculture, Hokkaido University, Kita-ku Kita 9 Nishi 9, Sapporo, 060-8589, Japan.
Abstract:
The effect of vitamin K on bovine endometrial epithelial cells has not been thoroughly investigated. The objective of this study was to examine the effect of the biologically active form of vitamin K, menaquinone-4, on gene expression in bovine endometrial epithelial cells. First, we examined the mRNA and protein expression levels of UBIAD1, a menaquinone-4 biosynthetic enzyme. Second, we screened for potential target genes of menaquinone-4 in bovine endometrial epithelial cells using RNA-sequencing. We found 50 differentially expressed genes; 42 were upregulated, and 8 were downregulated. Among them, a dose-dependent response to menaquinone-4 was observed for the top three upregulated (TRIB3, IL6, and TNFAIP3) and downregulated (CDC6, ORC1, and RRM2) genes. It has been suggested that these genes play important roles in reproductive events. In addition, GDF15 and VEGFA, which are important for cellular functions as they are commonly involved in pathways, such as positive regulation of cell communication, cell differentiation, and positive regulation of MAPK cascade, were upregulated in endometrial epithelial cells by menaquinone-4 treatment. To the best of our knowledge, this is the first study showing the expression of UBIAD1 in the bovine uterus. Moreover, the study determined menaquinone-4 target genes in bovine endometrial epithelial cells, which may positively affect pregnancy with alteration of gene expression in cattle uterus.
Insights
Menaquinone-4, a form of vitamin K, alters gene expression in bovine endometrial cells. This study identified key genes potentially impacting cattle reproduction and pregnancy outcomes.
Area of Science:
- Reproductive biology
- Nutritional science
- Molecular biology
Background:
- The role of vitamin K in bovine endometrial health is largely unexplored.
- Menaquinone-4 is the biologically active form of vitamin K, crucial for various cellular functions.
Purpose of the Study:
- To investigate the effects of menaquinone-4 on gene expression in bovine endometrial epithelial cells.
- To identify potential menaquinone-4 target genes involved in reproductive events in cattle.
Main Methods:
- Examined mRNA and protein expression of UBIAD1, a menaquinone-4 biosynthetic enzyme.
- Utilized RNA-sequencing to screen for menaquinone-4 target genes in bovine endometrial cells.
- Analyzed dose-dependent responses of specific upregulated and downregulated genes.
Main Results:
- Identified 50 differentially expressed genes (42 upregulated, 8 downregulated) in response to menaquinone-4.
- Observed dose-dependent changes in key genes like TRIB3, IL6, TNFAIP3, CDC6, ORC1, and RRM2.
- Found upregulation of GDF15 and VEGFA, involved in cell communication, differentiation, and MAPK signaling.
Conclusions:
- This is the first study to report UBIAD1 expression in the bovine uterus.
- Menaquinone-4 significantly alters gene expression in bovine endometrial cells, with potential benefits for cattle reproduction and pregnancy.
- Identified novel menaquinone-4 target genes that may influence uterine function and fertility.
