Ying NAN, Yifan XIE, Menting ZHU, Heng YANG, Zongsheng ZHAO

MicroRNA-200c Mediates the Mechanism of MAPK8 Gene Regulating Follicular Development in Sheep

Most sheep breeds are seasonal estrus and single birth animals. In order to improve pastoral area economy and lambing rate, In order toimprove pastoral area economy and lambing rate, and It is very important to study the key genes aff ecting sheep reproductive traits atthe molecular level. On the basis of early verification of the relationship between miR-200c and MAPK8 gene target. In this experiment,after over-expression of miR-200c in ovarian granulosa cells, the expression of mitogen-activated protein kinase 8 (MAPK8), frizzledclass receptor 3 (FZD3), G protein subunit alphaq (GNAQ), jun proto-oncogene (JUN), protein kinase C beta (PRKCB) in estrus-relatedpathway genes was detected by qRT-PCR, and the expression of estradio (E2) and progesterone (P4) in reproductive stimulation wasdetected by ELISA. The relationship between MAPK8 gene and follicular ovulation was analyzed. The results showed that up-regulatedcorpus follicular mRNA expressions of FZD3, GNAQ (P

MicroRNA-200c Koyunda MAPK8 Gen Düzenleyici Foliküler Gelişim Mekanizmasına Aracılık Eder

Koyun ırklarının çoğu mevsimsel östrus gösteren ve tek doğum yapan hayvanlardır. Kırsal alan ekonomisini iyileştirmek ve kuzulama oranını artırmak için, miR-200c ve MAPK8 gen hedefl eri arasındaki ilişkinin erken doğrulanması temelinde koyun üreme özelliklerini etkileyen önemli genleri moleküler düzeyde incelemek oldukça önemlidir. Bu çalışmada, yumurtalık granüloza hücrelerinde miR200c’nin aşırı ekspresyondan sonra, östrusla ilişkili yolak genlerinden Mitojen Aktiveli Protein Kinaz 8 (MAPK8), Frizzled sınıf reseptör 3 (FZD3), G Protein Altünite Alfa Q (GNAQ), Jun Protoonkogen (JUN), Protein Kinaz C Beta (PRKCB) ekspresyonu QT-PCR ile ve üreme stimülasyonunda östradiol (E2) ve progesteron (P4) ekspresyasyonu ELISA ile saptandı. MAPK8 geni ile foliküler ovulasyon arasındaki ilişki incelendi. Sonuçlar, FZD3, GNAQ’nın korpus foliküler mRNA ekspresyonlarının arttığını (P

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1. Zhang GX, Chu MX, Wang JY, Fang L, Ye SC: Polymorphism of exon 10 of prolactin receptor gene and its relationship with prolificacy of Jining Grey goats. Hereditas, 29, 329-336, 2007. DOI: 10.3321/j.issn:0253- 9772.2007.03.014

2. Rezaei R, Wu ZL, Hou YQ, Fuller WB, Wu GY: Amino acids and mammary gland development: Nutritional implications for milk production and neonatal growth. J Anim Sci Biotechnol, 7:20, 2016. DOI: 10.1186/ s40104-016-0078-8

3. Haziak K, Herman, AP, Wojtulewicz K, Pawlina B, Paczesna K, Bochenek J, Tomaszewska-Zaremba D: Effect of CD14/TLR4 antagonist on GnRH/LH secretion in ewe during central inflammation induced by intracerebroventricular administration of LPS. J Anim Sci Biotechnol, 9:52, 2018. DOI: 10.1186/s40104-018-0267-8

4. Huang HY, Li L, Ye HH, Feng B, Li S: Identification and distribution of gonadotropin-releasing hormone-like peptides in the brain of horseshoe crab Tachypleus tridentatus. Chin J Oceanol Limnol, 31, 384-390, 2013. DOI: 10.1007/s00343-013-2067-5

5. Ning HM, Ge YM, Su J, Zhang WL, Yao Y, Yang GH, Lei ZH: Effects of orexin A on mRNA expression of various neuropeptides in the hypothalamus and pituitary, and on serum LH levels in ovariectomized gilts. Agric Sci China, 9, 1362-1371, 2010. DOI: 10.1016/S1671-2927(09)60227-3

6. Fournet N, Weitsman SR, Zachow RJ, Magoffin DA: Transforming growth factor-beta inhibits ovarian 17 alpha-hydroxylase activity by a direct noncompetitive mechanism. Endocrinology, 137, 166-174, 1996. DOI: 10.1210/endo.137.1.8536609

7. Kang SK, Tai CJ, Cheng KW, Leung PCK: Gonadotropin-releasing hormone activates mitogen-activated protein kinase in human ovarian and placental cells. Mol Cell Endocrinol, 170, 143-151, 2000. DOI: 10.1016/ S0303-7207(00)00320-8

8. Patergnani S, Marchi S, Rimessi A, Bonora M, Giorgi C, Mehta KD, Pinton P: PRKCB/protein kinase C, beta and the mitochondrial axis as key regulators of autophagy. Autophagy, 9, 1367-1385, 2013. DOI: 10.4161/ auto.25239

9. Mugami S, Dobkin Bekman M, Rahamim Ben Navi L, Naor Z: Differential roles of PKC isoforms (PKCs) in GnRH stimulation of MAPK phosphorylation in gonadotrope derived cells. Mol Cell Endocrinol, 463, 97-105, 2018. DOI: 10.1016/j.mce.2017.04.004

10. An XP, Hou JX, Zhao HB, Li G, Bai L, Peng JY, Yan QM, Song YX, Wang JG, Cao BJ: Polymorphism identification in goat GnRH1 and GDF9 genes and their association analysis with litter size. Anim Genet, 44, 234- 238, 2013. DOI: 10.1111/j.1365-2052.2012.02394.x

11. An XP, Han P, Hou JX, Zhao HB, Yan Y, Ma T, Fang F, Meng FX, Song YX, Wang JG, Cao BY: Molecular cloning and characterization of KISS1 promoter and effect of KISS1 gene mutations on litter size in the goat. Genet Mol Res, 12, 4308-4316, 2013. DOI: 10.4238/2013.February.28.13

12. Carletti MZ, Christenson LK: Rapid effects of LH on gene expression in the mural granulosa cells of mouse periovulatory follicles. Reproduction, 137, 843-855, 2009. DOI: 10.1530/REP-08-0457

13. Ravasi T, Suzuki H, Pang KC, Katayama S, Furuno M, Okunishi R, Fukuda S, Ru K, Frith MC, Gongora MM, Grimmond SM, Hume DA, Hayashizaki Y, Mattick JS: Experimental validation of the regulated expression of large numbers of non-coding RNAs from the mouse genome. Genome Res, 16, 11-19, 2006. DOI: 10.1101/gr.4200206

14. Kuang YQ, Cao JX, Xu FF, Chen Y: Duplex-specific nucleasenucleasemediated amplification strategy for mass spectrometry quantification of miRNA-200c in breast cancer stem cells. Anal Chem, 91, 8820-8826, 2019. DOI: 10.1021/acs.analchem.8b04468

15. Zhang LQ, Lu N: Status and prospect of the role of miR-200c in early diagnosis of gastric cancer. World Chin J Digestol, 6, 382-388, 2019.

16. Zhang L, Wang K, Wu Q, Jin L, Lu H, Shi Y, Liu L, Yang L, Lv L: Let7 inhibits the migration and invasion of extravillous trophoblast cell via targeting MDM4. Mol Cell Probes, 45, 48-56, 2019. DOI: 10.1016/j. mcp.2019.05.002

17. Shi J, Ye Q, Zhou Z, Zhou XC, Di W: Expression of miRNA-200c in IIIc stage ovarian serous cystadenocarcinoma and its role in metastasis. Mod Trends Immunol, 1, 36-41, 2016.

18. Romerowicz-Misielak M, Koziorowski M: The gonadotropins subunits, GnRH and GnRH receptor gene expression and role of carbon monoxide in seasonal breeding animals. Ann Anim Sci, 12, 15-23, 2012. DOI: 10.2478/v10220-012-0002-x

19. Mélanie V, Olivier K, Nathalie H, Edith C, Yann LP, Yves C, Jean M, Porcher, Francois B: 17α-Ethinylestradiol and nonylphenol affect the development of for ANrain GnRH neurons through an estrogen receptors-dependent pathway. Reprod Toxicol, 33, 198-204, 2012. DOI: 10.1016/j.reprotox.2011.04.005

20. Barutcu SA, Girnius N, Vernia S, Davis RJ: Role of the MAPK/c-Jun NH2-terminal kinase signaling pathway in starvation-induced autophagy. Autophagy, 14, 1586-1595. 2018. DOI: 10.1080/15548627.2018.1466013

21. Wang Y, Bi Y, Zuo Q, Zhang W, Li D, He N, Cheng S, Zhang Y, Li B: MAPK8 regulates chicken male germ cell differentiation through JNK signaling pathway. J Cell Biochem, 119, 1548-1557, 2018. DOI: 10.1002/ jcb.26314

22. Zhang L, Fang Y, Cheng X, Lian YJ, Xu HL: Silencing of long noncoding RNA SOX21-AS1 relieves neuronal oxidative stress injury in mice with Alzheimer’s disease by upregulating FZD3/5 via the Wnt signaling pathway. Mol Neurobiol, 56, 3522-3537, 2019. DOI: 10.1007/ s12035-018-1299-y

23. Li J, Gao H, Tian Z, Wu Y, Wang Y, Fang Y, Lin L, Han Y, Wu S, Haq I, Zeng S: Effects of chronic heat stress on granulosa cell apoptosis and follicular atresia in mouse ovary. J Anim Sci Biotechnol, 7:57, 2016, DOI: 10.1186/s40104-016-0116-6

24. Wang MX, Shao XG: The serum progesterone level in superovulation was increased. Chin J Pract, 24, 1004-3845, 2015.

25. Nogueira DM, Cavalieri J, Fitzpatrick LA, Gummow B, Blache D, Parker AJ: Effect of hormonal synchronisation and/or short-term supplementation with maize on follicular dynamics and hormone profiles in goats during the non-breeding season. Anim Reprod Sci, 171, 87-97, 2016. DOI: 10.1016/j.anireprosci.2016.06.003