Hostname: page-component-586b7cd67f-rdxmf Total loading time: 0 Render date: 2024-11-22T15:09:22.390Z Has data issue: false hasContentIssue false
  • English
  • Français

Orexin A promotes progesterone secretion in luteinized granulose cells of Mongolian Ovis aries ovary by PRRT2 and ABCG1 genes

Published online by Cambridge University Press:  03 March 2021

Mengyuan Xie ,
Dong Han  and
Xiaojing Xu Open the ORCID record for Xiaojing Xu [Opens in a new window]
Mengyuan Xie
Affiliation:
Key Laboratory of Basic Veterinary Medicine, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, China
Dong Han
Affiliation:
Key Laboratory of Basic Veterinary Medicine, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, China
Xiaojing Xu*
Affiliation:
Key Laboratory of Basic Veterinary Medicine, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, China
*
Author for correspondence: Xiaojing Xu. Key Laboratory of Basic Veterinary Medicine, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, China. E-mail:[email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

To study the role of orexin A in the reproductive regulation of Mongolian sheep, ovine ovarian granulosa cells were cultured in vitro. The cells were divided into groups after luteinization, the experimental group was given orexin A and the transcriptome was sequenced together with that of the control group. The different genes related to reproduction were screened out. qRT-PCR, western blot and enzyme-linked immunosorbent assay (ELISA) were used to verify the selected genes and detect the effect on progesterone secretion. In total, 123 differentially expressed genes were obtained by sequencing. Six genes with high expression related to reproduction (PRRT2, ABCG1, SOX4, TBX3, ID1 and ATP8) were screened. The results of qRT-PCR were consistent with those of sequencing; western blot and ELISA were used to verify the protein levels of steroidogenic acute regulatory protein (StAR) and its related PRRT2 and ABCG1, and to detect their effect on progesterone secretion. Validation results were consistent with those of qRT-PCR and sequencing. The experimental group was given orexin A and compared with the control group. Expression of PRRT2 protein was significantly increased (P < 0.05), ABCG1 protein expression was significantly decreased (P < 0.05), StAR expression was significantly increased (P < 0.05), and progesterone secretion was significantly increased (P < 0.05). The results showed that orexin A promoted the expression of StAR by upregulating PRRT2 and downregulating ABCG1, therefore affecting secretion of progesterone. Gene expression characteristics of orexin A affecting progesterone secretion were preliminarily explored; this study provides a theoretical basis for further study on signalling pathways and reproductive regulation in Mongolian sheep.

Keywords

ABCG1Luteinized granulosa cellsOrexin AProgesteronePRRT2
Type
Research Article
Information
Zygote , Volume 29 , Issue 4 , August 2021 , pp. 286 - 292
Copyright
© The Author(s), 2021. Published by Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Belani, M, Deo, A, Shah, P, Banker, M, Singal, P and Gupta, S (2018). Differential insulin and steroidogenic signaling in insulin resistant and non-insulin resistant human luteinized granulosa cells – a study in PCOS patients. J Steroid Biochem Mol Biol 178, 283–92.CrossRefGoogle ScholarPubMed
Desquiret-Dumas, V, Clément, A, Seegers, V, Boucret, L, Ferré-L’Hotellier, V, Bouet, PE, Descamps, P, Procaccio, V, Reynier, P and May-Panloup, P (2017). The mitochondrial DNA content of cumulus granulosa cells is linked to embryo quality. Hum Reprod 32, 607–14.Google ScholarPubMed
Dobrzyn, K, Smolinska, N, Kiezun, M, Szeszko, K, Rytelewska, E, Kisielewska, K, Gudelska, M and Kaminski, T (2018). The in vitro effect of progesterone on the orexin system in porcine uterine tissues during early pregnancy. Acta Vet Scand 60, 76.CrossRefGoogle ScholarPubMed
Dobrzyn, K, Szeszko, K, Kiezun, M, Kisielewska, K, Rytelewska, E, Gudelska, M, Wyrebek, J, Bors, K, Kaminski, T and Smolinska, N (2019). In vitro effect of orexin A on the transcriptomic profile of the endometrium during early pregnancy in pigs. Anim Reprod Sci 200, 31–42.CrossRefGoogle ScholarPubMed
Groeneveled, LF, Lenstra, J.A, Eding, H, Toro, MA, Scherf, B, Pilling, D, Negrini, R, Finlay, EK, Jianlin, H, Groeneveld, E and Weigend, S (2010). Genetic diversity in farm animals – a review. Anim Genet 41, 6–31.CrossRefGoogle Scholar
Guo, Y and Cao, J (2012). The research progression of ABCA1 and ABCG1 in reverse cholesterol transport. Foreign Med Sci (Sect Medgeogr) 3, 213–7.Google Scholar
Guo, YX, Zhang, GM, Yao, XL, Tong, R, Cheng, CY, Zhang, TT, Wang, ST, Yang, H and Wang, F (2019). Effects of nitric oxide on steroidogenesis and apoptosis in goat luteinised granulosa cells. Theriogenology 126, 55–62.CrossRefGoogle Scholar
Ju, SJ, Zhao, YY, Chao, XC and Guo, L (2014). Orexin A protects cells from apoptosis by regulating FoxO1 and mTORC1 through the OX1R/PI3K/AKT signaling pathway in hepatocytes. Int J Mol Med 34, 153–9.CrossRefGoogle ScholarPubMed
Lehoux, J-G, Hales, DB, Fleury, A, Briere, N, Martel, D and Ducharme, L (1999). The in vivo effects of adrenocorticotropin and sodium restriction on the formation of different species of steroidogenic acute regulatory protein in rat adrenal. Endocrinology 140, 5154–64.CrossRefGoogle ScholarPubMed
Li, M, Zu, N, Zhang, C, Xu, X and Liu, Y (2017). Orexin A influence progesterone secreted and the content of cAMP in sheep ovarian granulosa cells. Vet Sci China 47, 1433–1440.Google Scholar
Li, M, Zu, N, Zhang, CS, Xie, MY, Liu, YZ and Xu, XJ (2019). Orexin A promotes granulosa cell secretion of progesterone in sheep. Iran J Vet Res 20, 136–42.Google ScholarPubMed
Lu, X (2011). Isolation, Culture and Identification of Luteal Cells from Goats. Yangling: Northwest A&F University.Google Scholar
Marathe, N, Rangaswami, H, Zhuang, S, Boss, GR and Pilz, RB (2012). Prosurvival effects of 17β-estradiol on osteocytes are mediated by nitric oxide/cGMP via differential actions of cGMP-dependent protein kinases I and II. J Biol Chem 287, 978–88.CrossRefGoogle Scholar
Spießberger, B, Bernhard, D, Herrmann, S, Feil, S, Werner, C, Luppa, PB and Hofmann, F (2009). cGMP-dependent protein kinase II and aldosterone secretion. FEBS J 276, 1007–13.CrossRefGoogle Scholar
Wang, Z, Huang, R, Pan, L, Li, X and Shi, F (2006). Molecular structures, physiological roles and regulatory mechanisms of cyclic nucleotide-gated ion channels. Chin J Biochem Mol Biol 22, 282–8.Google Scholar
Yang, T, Wang, S and Wang, J (2015). Recent advances of ABCG1 in cholesterol transport. J Med Postgrad 1, 89–93.Google Scholar
Yang, T, Wang, H, Tian, K and Hao, Z (2017). In vitro culture of porcine ovarian granulosa cells. Anim Sci Abroad – Pigs Poultry 37, 38–42.Google Scholar
Yu, W, Zhang, G, Song, X, Zhen, J, Chi, X and Wang, Z (2018). Effect of Lonicera caerulea anthocyanins on the expression of LDLR, ABCG1 and ABCA1 genes in livers of hyperlipidemic rats. Food Sci 39, 205–10.Google Scholar
Zu, Ning, Li, Min, Huang, Xiaoyu, Xu, Xiaojing, Mi, Yan and Liu, Yongzhi (2017). Effect of orexin A on the expression of progesterone related protein in granulosa cells of sheep ovary. Chin J Vet Sci 37, 1993–7.Google Scholar