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https://doi.org/10.5713/ab.22.0097    [Accepted] Published online June 30, 2022.
12-oxoeicosatetraenoic acid, a candidate of signal for placenta separation, activates matrix metalloproteinase and induces apoptosis in bovine trophoblast cells
Hachiro Kamada1,* 
1Institute of livestock and Grassland Science, NARO, Tsukuba, 305-0901, Japan
2Tohoku Agricultural Research Center, NARO, Akahira-4, Shimokuriyagawa, Morioka, 020-0198, Japan
Correspondence:  Hachiro Kamada, Tel: +81-019-643-3573, Fax: +81-019-643-3588, Email: kama8@affrc.go.jp
Received: 10 March 2022   • Revised: 26 May 2022   • Accepted: 22 June 2022
Abstract
Objective
12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid (12-KETE), a metabolite of arachidonic acid, is a strong candidate of signal for placenta separation following calf discharge at delivery. In the present study, the effects of 12-KETE on bovine trophoblast cells were investigated to determine its function in the placentome at delivery.
Methods
Bovine trophoblast cells derived from blastocysts were used. They were cocultured with or without fibroblasts derived from bovine placentome and/or bovine uterine epithelial cells. 12-KETE was added to the culture medium.
Results
Bovine trophoblast cells contained binucleate cells and strongly expressed CDX-2 genes. Addition of 12-KETE to the trophoblast cell colony without feeder cells or that on a fibroblast monolayer induced rapid exfoliation of the colony. After 12-KETE addition, trophoblast cells emitted strong fluorescence caused by the degradation of dye-quenched collagen, indicating that 12-KETE activated matrix metalloproteinase of the trophoblast cells. Exfoliated cell colonies were stained with YOPRO-1, but not propidium iodide. Moreover, DNA fragmentation and Bax gene (apoptosis stimulator) upregulation were observed in exfoliated cells, indicating that 12-KETE induced trophoblast cell apoptosis. These results were consistent with previous in vivo observations; however, even a lower concentration of 12-KETE activated trophoblast protease. Meanwhile, fibroblasts derived from the bovine placentome converted arachidonic acid to 12-KETE.
Conclusion
These observations indicate that 12-KETE may serve as a signal for placenta separation at delivery.
Keywords: Apoptosis; Bovine; Matrix Metalloproteinase; Trophoblast; 12-Oxoeicosatetraenoic Acid
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