克氏原螯虾前列腺素E<sub>2</sub>受体EP4基因的序列特征及表达分析

doi:10.16378/j.cnki.1003-1111.21033

Abstract
Figure/Table
References
Related Citation (15)

Download: PDF (6713 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Prostaglandin E₂ (PGE₂) receptor EP4 plays an important role in the regulation of ovarian development, ovulation and reproduction in animals. The cDNA sequence of the prostaglandin E₂ (PGE₂) receptor EP4 gene with the complete ORF region was obtained from the ovary of Procambarus clarkii using transcriptome sequencing. In order to explore the sequence characteristics and the function of EP4 gene in ovary of P. clarkii (PcEP4), the gene sequence was analyzed by bioinformatics, and the gene expression pattern was detected by real-time fluorescent quantitative PCR. Bioinformatics analysis results showed that the ORF of PcEP4 was 2505 bp, encoding a deduced protein with 835 amino acids; PcEP4 is a typical G protein coupled receptor with 7 transmembrane α-helical structures; EP4 amino acid sequence of P. clarkii and Litopenaeus vannamei were the most consistent,and the genetic relationship was the closest. Gene expression analysis results showed that PcEP4 gene expression level was the highest in the ovary in P. clarkii; in the Ⅰ—Ⅵ stage of ovaries in P. clarkii, PcEP4 gene expression level was the highest in stage Ⅴ ovary; At 36, 48 and 60 h after PGE₂ injection, the expression level of PcEP4 gene in the ovaries of P. clarkii was significantly higher than that in the control group (P<0.05), the egg holding rate of P. clarkii also increased compared with the control group at 72 h after PGE₂ injection. The above results indicate that, as a PGE₂ receptor, EP4 plays an important role in promoting oocyte maturation and ovulation of P. clarkii together with PGE₂.

Key words： Procambarus clarkii prostaglandin E₂ receptor EP4 gene expression ovary

Received: 01 March 2021

Q786

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	JIANG Hongxia
	ZHANG Ran
	LI Yizheng
	LIU Xuewei
	ZHANG Shuaishuai
	YU Miao
	WANG Lei
	ZHANG Meng
	QIAO Zhigang
	LI Xuejun

Cite this article:

JIANG Hongxia,ZHANG Ran,LI Yizheng, et al. Characterization and Expression Analyses of Prostaglandin E₂ Receptor EP4 Gene in Procambarus clarkii[J]. Fisheries Science, 2023, 42(4): 585-593.

URL:

https://www.shchkx.com/EN/10.16378/j.cnki.1003-1111.21033 OR https://www.shchkx.com/EN/Y2023/V42/I4/585

[1]CAUGHEY G E, CLELAND L G, PENGLIS P S, et al. Roles of cyclooxygenase (COX)-1 and COX-2 in prostanoid production by human endothelial cells:selective up-regulation of prostacyclin synthesis by COX-2[J].Journal of Immunology,2001,167(5):2831-2838.
[2]STANLEY D. Prostaglandins and other eicosanoids in insects:biological significance[J].Annual Review of Entomology,2006,51:25-44.
[3]TAHARA D, YANO I. Development of hemolymph prostaglandins assay systems and their concentration variations during ovarian maturation in the kuruma prawn, Penaeus japonicus[J].Aquaculture,2003,220(1/2/3/4):791-800.
[4]MEUNPOL O, DUANGJAI E, YOONPUN R, et al. Detection of prostaglandin E_2 in polychaete Perinereis sp.and its effect on Penaeus monodon oocyte development in vitro[J].Fisheries Science,2010,76(2):281-286.
[5]REDDY P S, REDDY P R, NAGARAJU G P C. The synthesis and effects of prostaglandins on the ovary of the crab Oziotelphusa senex senex[J].General and Comparative Endocrinology,2004,135(1):35-41.
[6]SUGIMOTO Y, NAMBA T, SHIGEMOTO R, et al. Distinct cellular localization of mRNAs for three subtypes of prostaglandin E receptor in kidney[J].The American Journal of Physiology,1994,266(5 Pt 2):F823-F828.
[7]KONGER R L, BILLINGS S D, THOMPSON A B, et al. Immunolocalization of low-affinity prostaglandin E2 receptors, EP1 and EP2, in adult human epidermis[J].Journal of Investigative Dermatology,2005,124(5):965-970.
[8]SEGI E, HARAGUCHI K, SUGIMOTO Y, et al. Expression of messenger RNA for prostaglandin E receptor subtypes EP4/EP2 and cyclooxygenase isozymes in mouse periovulatory follicles and oviducts during superovulation[J].Biology of Reproduction,2003,68(3):804-811.
[9]BAYNE R A L, EDDIE S L, COLLINS C S, et al. Prostaglandin E2 as a regulator of germ cells during ovarian development[J].The Journal of Clinical Endocrinology & Metabolism,2009,94(10):4053-4060.
[10]农业农村部渔业渔政管理局,全国水产技术推广总站,中国水产学会.2020中国渔业统计年鉴[M].北京:中国农业出版社,2020.
[11]李胜,赵维信.克氏原螫虾大颚器在卵巢发育周期中的组织结构变化[J].上海水产大学学报,1999,8(1):12-18.
[12]LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method[J].Methods,2001,25(4):402-408.
[13]SWETHA C H, GIRISH B P, HEMALATHA M, et al. Induction of vitellogenesis, methyl farnesoate synthesis and ecdysteroidogenesis in two edible crabs by arachidonic acid and prostaglandins[J].Journal of Experimental Biology,2020,223(3):jeb.212381.
[14]水燕,汤雪君,徐增洪,等.切除眼柄对加速克氏原螯虾卵巢发育进程的组织学研究[J].动物学杂志,2016,51(3):449-454.
[15]SUPRIYA N T, SUDHA K, KRISHNAKUMAR V, et al. Molt and reproduction enhancement together with hemolymph ecdysteroid elevation under eyestalk ablation in the female fiddler crab, Uca triangularis (Brachyura:Decapoda)[J].Chinese Journal of Oceanology and Limnology,2017,35(3):645-657.
[16]管政兵,水燕,周鑫,等.CaM与PMCA可能参与眼柄切除诱导克氏原螯虾卵巢成熟的早期分子过程[J].湖北农业科学,2012,51(21):4846-4849.
[17]黄智伟,石耀华,王爱民,等.眼柄摘除对红螯光壳螯虾生长、性腺发育及体色的影响[J].海洋渔业,2019,41(3):322-328.
[18]姜丽楠,闫红伟,罗明正,等.眼柄切除对口虾蛄卵巢、肝胰脏和肌肉生化组分的影响[J].水产科学,2018,37(4):441-447.
[19]UAWISETWATHANA U, LEELATANAWIT R, KLANCHUI A, et al. Insights into eyestalk ablation mechanism to induce ovarian maturation in the black tiger shrimp[J].PLoS One,2011,6(9):e24427.
[20]石宝通,钱照君,王辉,等.克氏原螯虾泛素结合酶E2基因的克隆及其在卵巢中的表达分析[J].上海海洋大学学报,2018,27(6):814-824.
[21]水燕,徐增洪,周鑫.Tropomyosin基因在克氏原螯虾(Procambarus clarkii)卵巢发育过程中的表达[J].安徽农业大学学报,2016,43(1):42-46.
[22]刘红,崔俊,颜婕,等.克氏原螯虾卵黄蛋白原受体cDNA部分序列的克隆及mRNA表达量的相对定量[J].渔业科学进展,2014,35(6):83-89.
[23]FINDLAY J, ELIOPOULOS E. Three-dimensional modelling of G protein-linked receptors[J].Trends in Pharmacological Sciences,1990,11(12):492-499.
[24]PROBST W C, SNYDER L A, SCHUSTER D I, et al. Sequence alignment of the G-protein coupled receptor superfamily[J].DNA and Cell Biology,1992,11(1):1-20.
[25]ATTWOOD T K, ELIOPOULOS E E, FINDLAY J B C. Multiple sequence alignment of protein families showing low sequence homology:a methodological approach using database pattern-matching discriminators for G-protein-linked receptors[J].Gene,1991,98(2):153-159.
[26]SPAZIANI E P, HINSCH G W, EDWARDS S C. Changes in prostaglandin E_2 and F₂α during vitellogenesis in the Florida crayfish Procambarus paeninsulanus[J].Journal of Comparative Physiology B,1993,163(7):541-545.
[27]PREECHAPHOL R, KLINBUNGA S, KHAMNAMT-ONG B, et al. Isolation and characterization of genes functionally involved in ovarian development of the giant tiger shrimp Penaeus monodon by suppression subtractive hybridization (SSH)[J].Genetics and Molecular Biology,2010,33(4):676-685.