内分泌调节基因在三倍体雌性虹鳟不同发育阶段的表达分析
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摘要
为探究内分泌关键调节基因在不同倍性雌性虹鳟Oncorhynchus mykiss早期和后期发育不同阶段的表达模式,采用Real-time PCR方法,分别对二、三倍体雌性虹鳟早期发育阶段(31～68 days post fertilization, dpf)脑组织和不同发育阶段(160～450 dpf)性腺组织中促性腺激素释放激素(GnRH1)、促性腺激素释放激素受体(GnRHr)和促性腺激素α亚基(GTHa)基因的表达模式进行研究。结果表明:在早期发育阶段,三倍体雌性虹鳟脑中gnrh1和gnrhr的表达在某些时期呈显著上升(P<0.05),而gtha基因的表达显著降低(P<0.05),且始终维持在较低水平;三倍体雌性虹鳟在180～270 dpf时期,性腺组织中这3个基因的表达量均较检测初期(160 dpf)出现了显著下降(P<0.05),在发育后期360～450 dpf,这3个基因的表达量均较检测初期(160 dpf)下降了90%以上(P<0.05)。研究表明,三倍体雌性虹鳟腺垂体在早期发育阶段分泌促性腺激素的能力出现障碍,在三倍体雌性虹鳟的发育过程中内分泌生殖轴下丘脑—腺垂体—性腺受到阻断,这是导致其性腺发育异常的关键原因之一。
The expressions of gnrh1(Gonadotropin-releasing hormone 1), gnrhr(Gonadotropin-releasing hormone receptor) and gtha(Gonadotropin hormone α subunit) were detected in brains of diploid and triploid female rainbow trout Oncorhynchus mykiss(RBT) from 31 days post fertilization(dpf) to 68 dpf, and in gonads of diploid and triploid female RBT from 160 dpf to 450 dpf by Real-time PCR to explore the expressive patterns of pivotal endocrine genes in different ploidy female RBT at various developmental stages. It was found that the expression levels of gnrh1 and gnrhr were significantly elevated in brains of triploid female RBT in early developmental period(P<0.05). However, the expression levels of gtha was found to be significantly low in brains of triploid female RBT(P<0.05), remaining at very low level during the whole test period. There were significantly lower expression levels of gnrh1, gnrhr and gtha genes in gonads during the period of 180-270 dpf than those in initial detection(160 dpf)(P<0.05), being decreased by more than 90% during the period of 360-450 dpf compared with those in initial detection. The findings indicate that the ability to secrete gonadotropin hormone by adenohypophysis in pituitary gland, endocrine hypothalamus-pituitary-gonad(HPG) axis, is hindered in triploid female RBT during early developmental period, which is one of the prime cause of the gonads infertility in triploid female RBT.
The expressions of gnrh1(Gonadotropin-releasing hormone 1), gnrhr(Gonadotropin-releasing hormone receptor) and gtha(Gonadotropin hormone α subunit) were detected in brains of diploid and triploid female rainbow trout Oncorhynchus mykiss(RBT) from 31 days post fertilization(dpf) to 68 dpf, and in gonads of diploid and triploid female RBT from 160 dpf to 450 dpf by Real-time PCR to explore the expressive patterns of pivotal endocrine genes in different ploidy female RBT at various developmental stages. It was found that the expression levels of gnrh1 and gnrhr were significantly elevated in brains of triploid female RBT in early developmental period(P<0.05). However, the expression levels of gtha was found to be significantly low in brains of triploid female RBT(P<0.05), remaining at very low level during the whole test period. There were significantly lower expression levels of gnrh1, gnrhr and gtha genes in gonads during the period of 180-270 dpf than those in initial detection(160 dpf)(P<0.05), being decreased by more than 90% during the period of 360-450 dpf compared with those in initial detection. The findings indicate that the ability to secrete gonadotropin hormone by adenohypophysis in pituitary gland, endocrine hypothalamus-pituitary-gonad(HPG) axis, is hindered in triploid female RBT during early developmental period, which is one of the prime cause of the gonads infertility in triploid female RBT.
引文
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[21] Ji K,Hong S,Kho Y,et al.Effects of bisphenol S exposure on endocrine functions and reproduction of zebrafish[J].Environ Sci Technol,2013,47(15):8793-8800.
[22] Yang Xiaoxi,Liu Yuchen,Li Jia,et al.Exposure to bisphenol AF disrupts sex hormone levels and vitellogenin expression in zebrafish[J].Environ Toxicol,2016,31(3):285-294.
[23] Huang Tianqing,Sun Huizhi,Wang Yanna,et al.Effect of follicle cell autophagy on gonadal development of triploid female rainbow trout (Oncorhynchus mykiss)[J].Fish Physiol Biochem,2018,44(1):185-196.
[24] Espigares F,Carrillo M,Gómez A,et al.The forebrain-midbrain acts as functional endocrine signaling pathway of Kiss2/Gnrh1 system controlling the gonadotroph activity in the teleost fish European sea bass (Dicentrarchus labrax)[J].Biol Reprod,2015,92(3):70.
[2] Sumpter J P,Lincoln R F,Bye V J,et al.Plasma growth hormone levels during sexual maturation in diploid and triploid rainbow trout (Oncorhynchus mykiss)[J].Gen Comp Endocrinol,1991,83(1):103-110.
[3] Piferrer F,Beaumont A,Falguiere J C,et al.Polyploid fish and shellfish:production,biology and applications to aquaculture for performance improvement and genetic containment[J].Aquaculture,2009,293(3-4):125-156.
[4] Nagahama Y,Kobayashi T,Matsuda M.Sex determination,gonadal sex differentiation and sex change in fish[J].Tanpakushitsu Kakusan Koso.Protein,Nucleic Acid,Enzyme,2004,49(2):116-123.
[5] Avtanski D,Novaira H J,Wu Sheng,et al.Both estrogen receptor α and β stimulate pituitary GH gene expression[J].Mol Endocrinol,2014,28(1):40-52.
[6] Busby E R,Soeta S,Sherwood N M,et al.Molecular analysis of the koala reproductive hormones and their receptors:gonadotrophin-releasing hormone (GnRH),follicle-stimulating hormone β and luteinising hormone β with localisation of GnRH[J].J Neuroendocrinol,2014,26(12):870-887.
[7] Dickey J T,Swanson P.Effects of salmon gonadotropin-releasing hormone on follicle stimulating hormone secretion and subunit gene expression in Coho salmon (Oncorhynchus kisutch)[J].Gen Comp Endocrinol,2000,118(3):436-449.
[8] Bhandari R K,Taniyama S,Kitahashi T,et al.Seasonal changes of responses to gonadotropin-releasing hormone analog in expression of growth hormone/prolactin/somatolactin genes in the pituitary of masu salmon[J].Gen Comp Endocrinol,2003,130(1):55-63.
[9] Breen K M,Thackray V G,Hsu T,et al.Stress levels of glucocorticoids inhibit LHβ-subunit gene expression in gonadotrope cells[J].Mol Endocrinol,2012,26(10):1716-1731.
[10] Angelova K,Puett D.Differential responses of an invariant region in the ectodomain of three glycoprotein hormone receptors to mutagenesis and assay conditions[J].Endocrine,2002,19(2):147-154.
[11] 相福生,黄天晴,谷伟,等.硬骨鱼脑芳香化酶的表达调节、功能推测以及类固醇合成酶的研究进展[J].水产学杂志,2018,31(1):42-51.
[12] Wang Yaqin,Li Yingwen,Chen Qiliang,et al.Diethylstilbestrol impaired oogenesis of yellow catfish juveniles through disrupting hypothalamic-pituitary-gonadal axis and germ cell development[J].J Appl Toxicol,2018,38(3):308-317.
[13] Jeng S R,Yueh W S,Pen Y T,et al.Neuroendocrine gene expression reveals a decrease in dopamine D2B receptor with no changes in GnRH system during prepubertal metamorphosis of silvering in wild Japanese eel[J].Gen Comp Endocrinol,2014,206:8-15.
[14] Valcarce D G,Vuelta E,Robles V,et al.Paternal exposure to environmental 17-alpha-ethinylestradiol concentrations modifies testicular transcription,affecting the sperm transcript content and the offspring performance in zebrafish[J].Aquat Toxicol,2017,193:18-29.
[15] 闫浩,梁利群,常玉梅,等.遗传和温度因素对鱼类性别分化相关基因表达及性别比例的影响[J].大连海洋大学学报,2017,32(1):111-118.
[16] Ramachandra R K,Salem M,Gahr S,et al.Cloning and characterization of microRNAs from rainbow trout (Oncorhynchus mykiss):their expression during early embryonic development[J].BMC Dev Biol,2008,8:41.
[17] Park J W,Jin Y H,Oh S Y,et al.Kisspeptin2 stimulates the HPG axis in immature Nile tilapia (Oreochromis niloticus)[J].Comp Biochem Physiol Part B:Biochem Mol Biol,2016,202:31-38.
[18] Yang Qian,Yang Xianhai,Liu Jining,et al.Effects of BPF on steroid hormone homeostasis and gene expression in the hypothalamic—pituitary—gonadal axis of zebrafish[J].Environ Sci Pollut Res,2017,24(26):21311-21322.
[19] Filby A L,Van Aerle R,Duitman J,et al.The kisspeptin/gonadotropin-releasing hormone pathway and molecular signaling of puberty in fish[J].Biol Reprod,2008,78(2):278-289.
[20] Liu Xiaoshan,Ji K,Jo A,et al.Effects of TDCPP or TPP on gene transcriptions and hormones of HPG axis,and their consequences on reproduction in adult zebrafish (Danio rerio)[J].Aquat Toxicol,2013,134-135:104-111.
[21] Ji K,Hong S,Kho Y,et al.Effects of bisphenol S exposure on endocrine functions and reproduction of zebrafish[J].Environ Sci Technol,2013,47(15):8793-8800.
[22] Yang Xiaoxi,Liu Yuchen,Li Jia,et al.Exposure to bisphenol AF disrupts sex hormone levels and vitellogenin expression in zebrafish[J].Environ Toxicol,2016,31(3):285-294.
[23] Huang Tianqing,Sun Huizhi,Wang Yanna,et al.Effect of follicle cell autophagy on gonadal development of triploid female rainbow trout (Oncorhynchus mykiss)[J].Fish Physiol Biochem,2018,44(1):185-196.
[24] Espigares F,Carrillo M,Gómez A,et al.The forebrain-midbrain acts as functional endocrine signaling pathway of Kiss2/Gnrh1 system controlling the gonadotroph activity in the teleost fish European sea bass (Dicentrarchus labrax)[J].Biol Reprod,2015,92(3):70.