Cloning of Yak PIK3CB Gene and Its Expression During Follicular Development

doi:10.7668/hbnxb.20191376

Abstract

Abstract: The aim of this study was to investigate the sequence characteristics of phosphatidylinositol-3 kinase catalytic subunit β(PIK3CB/P110β) gene and compare its expression profile in different developmental stages of yak follicles.The sequence of PIK3CB gene was cloned from yak ovary tissue by RT-qPCR and analyzed by bioinformatics. The expression level of PIK3CB gene in different tissues of yak was detected by RT-qPCR method. The collected yak follicles were divided into three groups according to their diameter:large(≥ 7 mm), medium(3.0-6.9 mm) and small(≤ 2.9 mm). The total RNA was extracted from the oocytes and parietal granulosa cells of each group. Then, the relative expression of PIK3CB gene mRNA was detected by RT-qPCR.The results showed that the full-length CDS region of yak PIK3CB gene was 3 213 bp, encoding 1 070 amino acids. Bioinformatics analysis showed that PIK3CB protein was a hydrophilic acidic protein with no transmembrane structure and signal peptide, and the secondary structure was mainly composed of α-helix and random coil. The nucleotide homology and genetic tree analysis showed that yak PIK3CB gene was closely related to that of Bos mutus and Bos taurus. PIK3CB gene was expressed in heart, lung, kidney, liver, small intestine, stomach and muscle of yak, especially in spleen, uterus and ovary, the expression level was significantly higher than that in other tissues(P<0.05). The results of RT-qPCR showed that PIK3CB gene was expressed during follicular development, and in different follicular wall granulosa cells, the expression of mRNA increased with the development of follicles, and the expression in large and intermediate follicles was significantly higher than that in small follicles (P<0.01), but there was no significant difference in mRNA expression in oocytes(P>0.05).The results suggested that PIK3CB gene was involved in the regulation of yak follicular development, and it was one of the indispensable catalytic subunits of PI3K signal in regulating the function of granulosa cells. This study provides basic data for further study on the mechanism of PI3K-AKT signaling pathway in ovarian development.

Key words: Yak, PIK3CB, Tissue expression, Follicle

CLC Number:

S823.8+5
Q78

MA Hongcheng, XIONG Xianrong, WANG Han, HAI Zhuo, MIN Xingyu, LI Jian. Cloning of Yak PIK3CB Gene and Its Expression During Follicular Development[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(1): 210-218. doi: 10.7668/hbnxb.20191376.

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References

[1] 秦文昌,殷实,李泽沛,王斌,杨柳青,周婧雯,李键. 牦牛 HDAC8 基因特征及其在组织和卵母细胞成熟过程中的表达分析[J]. 华北农学报,2020,35(2):210-216. doi:10.7668/hbnxb.20190532.
Qin W C,Yin S,Li Z P,Wang B,Yang L Q,Zhou J W,Li J. The characteristic of yak HDAC8 gene and analysis of their expression in different tissues and during the oocyte maturation[J]. Acta Agriculturae Boreali-Sinica,2020,35(2):210-216.
[2] Sakaguchi K, Yanagawa Y, Yoshioka K, Suda T, Katagiri S, Nagano M. Relationships between the antral follicle count,steroidogenesis,and secretion of follicle-stimulating hormone and anti-Mü llerian hormone during follicular growth in cattle[J]. Reproductive Biology and Endocrinology,2019,17(1):88. doi:10.1186/s12958-019-0534-3.
[3] Liu Y X,Zhang Y,Li Y Y,Liu X M,Wang X X,Zhang C L,Hao C F,Deng S L. Regulation of follicular development and differentiation by intra-ovarian factors and endocrine hormones[J]. Frontiers in Bioscience-Landmark,2019,24:983-993.doi:10.2741/4763.
[4] Xu S R,Wei P,Yang Q L,Jia G X,Ma S K,Yang Q E,Jun Z,Zhang R N. Transcriptome analysis revealed key signaling networks regulating ovarian activities in the domestic yak[J]. Theriogenology,2020,147:50-56. doi:10.1016/j.theriogenology.2020.02.023.
[5] Bilanges B,Posor Y,Vanhaesebroeck B. PI3K isoforms in cell signalling and vesicle trafficking[J]. Nature Reviews Molecular Cell Biology,2019,20(9):515-534. doi:10.1038/s41580-019-0129-z.
[6] Dou Z X,Chattopadhyay M,Pan J A,Guerriero J L,Jiang Y P,Ballou L M,Yue Z Y,Lin R Z,Zong W X. The class IA phosphatidylinositol 3-kinase p110β subunit is a positive regulator of autophagy[J]. Journal of Cell Biology,2010,191(4):827-843. doi:10.1083/jcb.201006056.
[7] Dornan G L,Stariha J T B,Rathinaswamy M K,Powell C J,Boulanger M J,Burke J E. Defining how oncogenic and developmental mutations of PIK3R1 alter the regulation of Class IA phosphoinositide 3-kinases[J]. Structure,2019,28(2):145-156. doi:10.1016/j.str.2019.11.013.
[8] Salloum G,Jakubik C T,Erami Z,Heitz S D,Bresnick A R,Backer J M. PI3Kβ is selectively required for growth factor-stimulated macropinocytosis[J]. Journal of Cell Science,2019,132(16):231639. doi:10.1242/jcs.231639.
[9] Jiang N N,Dai Q J,Su X R,Fu J J,Feng X C,Peng J. Role of PI3K/AKT pathway in cancer:the framework of malignant behavior[J]. Molecular Biology Reports,2020,47:4587-4629. doi:10.1007/s11033-020-05435-1.
[10] Zheng W J,Nagaraju G,Liu Z L,Liu K. Functional roles of the phosphatidylinositol 3-kinases(PI3Ks) signaling in the mammalian ovary[J]. Molecular and Cellular Endocrinology,2012,356(1-2):24-30. doi:10.1016/j.mce.2011.05.027.
[11] Reddy P,Liu L,Adhikari D,Jagarlamudi K,Rajareddy S,Shen Y,Du C,Tang W L,Hämäläinen T,Peng S L,Lan Z J,Cooney A J,Huhtaniemi I,Liu K. Oocyte-specific deletion of Pten causes premature activation of the primordial follicle pool[J]. Science,2008,319(5863):611-613. doi:10.1126/science.1152257.
[12] 李倩. PI3K p110delta催化亚基在小鼠卵巢卵泡生长中的功能研究[D]. 南京:南京医科大学,2013.
Li Q. The p110delta Isoform of phosphoinositide 3-kinase catalytic subunits is key for hormonally stimulated ovarian follicle growth[D].Nanjing:Nanjing Medical University,2013.
[13] Yao H,Zhang J W,Wen J,Wang Y B,Niu W B,Teng Z,Zhao T T,Dai Y L,Zhang Y,Wang C,Qin Y Y,Xia G L,Zhang H.CDC42 controls the activation of primordial follicles by regulating PI3K signaling in mouse oocytes[J]. Bmc Biology,2018,16(1):73. doi:10.1186/s12915-018-0541-4.
[14] Zhang L,Li Y D,Wang Q C,Chen Z,Li X Y,Wu Z X,Hu C H,Liao D,Zhang W,Chen Z S. The PI3K subunits,P110α and P110β are potential targets for overcoming P-gp and BCRP-mediated MDR in cancer[J]. Molecular Cancer,2020,19(1):10. doi:10.1186/s12943-019-1112-1.
[15] Zhang Z,Zhang L Y,Wang B,Wei R,Wang Y X,Wan J P,Zhang C,Zhao L,Zhu X X,Zhang Y H,Chu C,Guo Q,Yin X Q,Li X. MiR-337-3p suppresses proliferation of epithelial ovarian cancer by targeting PIK3CA and PIK3CB[J]. Cancer Letters,2020,469:54-67. doi:10.1016/j.canlet.2019.10.021.
[16] Yang J,Nie J,Ma X L,Wei Y Q,Peng Y,Wei X W. Targeting PI3K in cancer:mechanisms and advances in clinical trials[J]. Molecular Cancer,2019,18(1):26. doi:10.1186/s12943-019-0954-x.
[17] John G B,Gallardo T D,Shirley L J,Castrillon D H. Foxo3 is a PI3K-dependent molecular switch controlling the initiation of oocyte growth[J]. Developmental Biology,2008,321(1):197-204. doi:10.1016/j.ydbio.2008.06.017.
[18] Jagarlamudi K,Liu L,Adhikari D,Reddy P,Idahl A,Ottander U,Lundin E,Liu K. Oocyte-specific deletion of Pten in mice reveals a stage-specific function of PTEN/PI3K signaling in oocytes in controlling follicular activation[J]. PLoS One,2009,4(7):e6186. doi:10.1371/journal.pone.0006186.
[19] Han S J,Vaccari S,Nedachi T,Andersen C B,Kovacina K S,Roth R A,Conti M. Protein kinase B/Akt phosphorylation of PDE3A and its role in mammalian oocyte maturation[J]. Embo Journal,2006,25(24):5716-5725. doi:10.1038/sj.emboj.7601431.
[20] Zheng W J,Liu K. The emerging role of maternal phosphatidylinositol 3 kinase(PI3K) signaling in manipulating mammalian preimplantation embryogenesis[J]. Cell Cycle,2011,10(2):178-179. doi:10.4161/cc.10.2.14475.
[21] Cantley L C. The phosphoinositide 3-kinase pathway[J]. Science,2002,296(5573):1655-1657. doi:10.1126/science.296.5573.1655.
[22] Dornan G L,Burke J E. Molecular mechanisms of human disease mediated by oncogenic and primary immunodeficiency mutations in Class IA phosphoinositide 3-kinases[J]. Frontiers in Immunology,2018,9:575. doi:10.3389/fimmu.2018.00575.
[23] Bi L, Okabe I, Bernard D J, Nussbaum R L. Early embryonic lethality in mice deficient in the p110β catalytic subunit of PI 3-kinase[J]. Mammalian Genome,2002,13(3):169-172. doi:10.1007/BF02684023.
[24] Kulkarni S,Sitaru C,Jakus Z,Anderson K E,Damoulakis G,Davidson K,Hirose M,Juss J,Oxley D,Chessa T A M,Ramadani F,Guillou H,Segonds-Pichon A,Fritsch A,Jarvis G E,Okkenhaug K,Ludwig R,Zillikens D,Mocsai A,Vanhaesebroeck B,Stephens L R,Hawkins P T. PI3K plays a critical role in neutrophil activation by immune complexes[J]. Science Signaling,2011,4(168):ra23-ra23. doi:10.1126/scisignal.2001617.
[25] Sato E. Intraovarian control of selective follicular growth and induction of oocyte maturation in mammals[J]. Proceedings of the Japan Academy Series B-physical and Biological Sciences,2015,91(3):76-91. doi:10.2183/pjab.91.76.
[26] Hunt P A,Hassold T J. Human female meiosis:what makes a good egg go bad?[J]. Trends in Genetics,2008,24(2):86-93. doi:10.1016/j.tig.2007.11.010.
[27] Shimizu T. Molecular and cellular mechanisms for the regulation of ovarian follicular function in cows[J]. Journal of Reproduction and Development,2016,62(4):323-329. doi:10.1262/jrd.2016-044.
[28] Gonzalez-Robayna I J, Falender A E, Ochsner S, Firestone G L, Richards J S. Follicle-stimulating hormone(FSH) stimulates phosphorylation and activation of protein kinase B(PKB/Akt) and serum and glucocorticoid-induced kinase(Sgk):Evidence for a kinase-independent signaling by FSH in granulosa cells[J]. Molecular Endocrinology,2000,14(8):1283-1300. doi:10.1210/mend.14.8.0500.
[29] Fan H Y, Liu Z L, Cahill N, Richards J S. Targeted disruption of Pten in ovarian granulosa cells enhances ovulation and extends the life span of luteal cells[J]. Molecular Endocrinology,2008,22(9):2128-2140. doi:10.1210/me.2008-0095.
[30] Guillermet-Guibert J,Bjorklof K,Salpekar A,Gonella C,Ramadani F,Bilancio A,Meek S,Smith A J H,Okkenhaug K,Vanhaesebroeck B. The p110β isoform of phosphoinositide 3-kinase signals downstream of G protein-coupled receptors and is functionally redundant with p110γ[J]. Proceedings of the National Academy of Sciences of the United States of America,2008,105(24):8292-8297. doi:10.1073/pnas.0707761105.
[31] Law N C,Hunzicker-Dunn M E. Insulin receptor substrate 1:the hub linking follicle-stimulating hormone to phosphatidylinositol-3 kinase activation[J]. Journal of Biological Chemistry,2016,291(9):4547-4560. doi:10.1074/jbc.M115.698761.
[32] Zheng W J, Gorre N, Shen Y, Noda T, Ogawa W, Lundin E, Liu K. Maternal phosphatidylinositol 3-kinase signalling is crucial for embryonic genome activation and preimplantation embryogenesis[J]. Embo Reports,2010,11(11):890-895. doi:10.1038/embor.2010.144.

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