Mitogen-activated protein kinase kinase kinase 8 (MAP3K8) mediates the LH-induced stimulation of progesterone synthesis in the porcine corpus luteum
Di Zhang A , Ying Liu A , Yan Cui B C and Sheng Cui A CA State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100094, PR China.
B The 306th Hospital of People’s Liberation Army, Beijing, 100101, PR China.
C Corresponding authors. Emails: cuisheng@cau.edu.cn; dryancui@163.com
Reproduction, Fertility and Development 31(9) 1444-1456 https://doi.org/10.1071/RD18478
Submitted: 27 June 2018 Accepted: 25 February 2019 Published: 1 May 2019
Abstract
Progesterone (P4) synthesized by the corpus luteum (CL) plays a key role in the establishment and maintenance of pregnancy. The LH signal is important for luteinisation and P4 synthesis in pigs. In a previous study, we demonstrated that mitogen-activated protein kinase kinase kinase 8 (MAP3K8) regulates P4 synthesis in mouse CL, but whether the function and mechanism of MAP3K8 in the pig is similar to that in the mouse is not known. Thus, in the present study we investigated the effects of MAP3K8 on porcine CL. Abundant expression of MAP3K8 was detected in porcine CL, and, in pigs, MAP3K8 expression was higher in mature CLs (or those of the mid-luteal phase) than in regressing CLs (late luteal phase). Further functional studies in cultured porcine luteal cells showed that P4 synthesis and the expression of genes encoding the key enzymes in P4 synthesis are significantly reduced when MAP3K8 is inhibited with the MAP3K8 inhibitor Tpl2 kinase inhibitor (MAP3K8i, 10 μM). After 12–24 h treatment of luteal cells with 100 ng mL−1 LH, MAP3K8 expression and P4 secretion were significantly upregulated. In addition, the 10 μM MAP3K8 inhibitor blocked the stimulatory effect of LH on P4 synthesis and extracellular signal-regulated kinase (ERK) 1/2 phosphorylation in porcine luteal cells. The LH-induced increases in MAP3K8 phosphorylation and expression, ERK1/2 phosphorylation and P4 synthesis were all blocked when protein kinase A was inhibited by its inhibitor H89 (20 μM) in porcine luteal cells. In conclusion, MAP3K8 mediates the LH-induced stimulation of P4 synthesis through the PKA/mitogen-activated protein kinase signalling pathway in porcine CL.
Additional keywords: extracellular signal-regulated kinase 1/2, pig, protein kinase A.
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