17α-甲基睾酮对草鱼性腺发育及性类固醇激素水平的影响
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摘要
利用养殖草鱼雌性个体所表现出的生长优势,探讨通过性逆转培育全雌草鱼品种的方法,具有较好的应用前景。实验采用拌料投喂诱导方式,在饲料中设置0、50、100和200 mg/kg 4个浓度梯度的17α-甲基睾酮(MT),连续投喂150 d,观测不同处理水平草鱼性腺结构和血液中类固醇激素雌二醇(E2)、睾酮(T)和11-酮基睾酮(11-KT)含量的变化规律,探讨草鱼性逆转过程中的特征变化。性腺的组织切片结果显示,从投喂30 d开始,各处理组卵巢中开始出现精原细胞。30～150 d,50 mg/kg MT处理组草鱼卵母细胞的生成及成熟受到抑制,发育迟缓;100 mg/kg MT处理组的生殖细胞受抑制程度要高于50 mg/kg MT处理组,实验结束时性腺已被大量精原细胞占据,有向精巢发育的趋势;200 mg/kg MT处理组精卵兼性状态较50 mg/kg MT处理组明显,但精原细胞的数量少于100 mg/kg MT处理组,对照组卵巢未出现精原细胞。血清类固醇激素测定结果显示,投喂不同剂量的MT,E_2浓度均显著升高,于45 d时达到最高值,之后趋于平稳下降,但仍显著高于对照组,未经处理的雄性草鱼,随着早期精巢的发育,其E_2水平也逐渐升高。随着MT投喂时间的延长,各处理组T浓度分别有不同程度的降低,45 d时各处理组降幅最大,且与MT浓度呈正相关,11-KT则有不同程度升高。研究表明,100 mg/kg MT为其性逆转较适宜的浓度,MT可使早期雌性草鱼血清E_2升高,发挥雄激素效应,从而促进卵巢卵细胞退化,精原细胞增殖。
By using the growth superiority demonstrated by the female individuals of the cultured grass carp, the method of breeding the whole female grass carp species by sex reversal is discussed, which has a good application prospect. The 17α-methyltestosterone(17α-MT) was selected to induce female-to-male as the male hormone, and set 4 concentration gradients evenly mixed with feed, i.e. 0 mg/kg, 50 mg/kg, 100 mg/kg, 200 mg/kg, which were respectively fed for 150 days. The objective of this study was to investigate the effects of different MT-treated diet on gonadal development and the changes of estradiol-17β(E2), testosterone(T) and 11-ketotestosterone(11-KT),and further the characteristics in the process of grass carp sex reversal. The morphological changes of grass carp were investigated by observing the changes of gonadal structure and serum steroid hormone content at different treatment levels to explore the sex reversal. Histological sections of gonads results showed that spermatogonia began to appear in the ovaries of all treatment groups from the 30 th day after feeding. With the prolongation of feeding time, 50 mg/kg MT treatment group can inhibit the formation and maturation of oocytes, resulting in developmental delay; The inhibition of germ cells in 100 mg/kg MT treatment group was higher than that in50 mg/kg MT treatment group, the gonads had been occupied by a large number of spermatogonia, and there was a tendency of developing testis at 210 d. The ovaries-testes state of 200 mg/kg MT treatment group at 210 d was more obvious than 50 mg/kg MT treatment group, but the number of spermatogonia was less than 100 mg/kg MT treatment group, while the control group ovary did not exhibit spermatogonia. The results of serum steroid hormone revealed that E_2 increased significantly which to some extent depended on the MT dosages, reached the highest value at 45 days and then decreased steadily, but was still significantly higher than that of the control group, however. With the development of early stage testis, E_2 level of control male also gradually increased. With the prolongation of MT feeding time, the concentrations of T in each treatment group decreased to different extents, and the biggest drop occurred in all treatment groups at 45 days, which was positively correlated with the MT concentration. The 11-KT increased to varying degrees. The results show that: 100 mg/kg was more suitable concentration to bring about sex reversal, MT can cause the early female fish serum E_2 to increase, and exert the effect of androgen, thereby promoting ovarian egg cell degradation, and spermatogonia proliferation.
By using the growth superiority demonstrated by the female individuals of the cultured grass carp, the method of breeding the whole female grass carp species by sex reversal is discussed, which has a good application prospect. The 17α-methyltestosterone(17α-MT) was selected to induce female-to-male as the male hormone, and set 4 concentration gradients evenly mixed with feed, i.e. 0 mg/kg, 50 mg/kg, 100 mg/kg, 200 mg/kg, which were respectively fed for 150 days. The objective of this study was to investigate the effects of different MT-treated diet on gonadal development and the changes of estradiol-17β(E2), testosterone(T) and 11-ketotestosterone(11-KT),and further the characteristics in the process of grass carp sex reversal. The morphological changes of grass carp were investigated by observing the changes of gonadal structure and serum steroid hormone content at different treatment levels to explore the sex reversal. Histological sections of gonads results showed that spermatogonia began to appear in the ovaries of all treatment groups from the 30 th day after feeding. With the prolongation of feeding time, 50 mg/kg MT treatment group can inhibit the formation and maturation of oocytes, resulting in developmental delay; The inhibition of germ cells in 100 mg/kg MT treatment group was higher than that in50 mg/kg MT treatment group, the gonads had been occupied by a large number of spermatogonia, and there was a tendency of developing testis at 210 d. The ovaries-testes state of 200 mg/kg MT treatment group at 210 d was more obvious than 50 mg/kg MT treatment group, but the number of spermatogonia was less than 100 mg/kg MT treatment group, while the control group ovary did not exhibit spermatogonia. The results of serum steroid hormone revealed that E_2 increased significantly which to some extent depended on the MT dosages, reached the highest value at 45 days and then decreased steadily, but was still significantly higher than that of the control group, however. With the development of early stage testis, E_2 level of control male also gradually increased. With the prolongation of MT feeding time, the concentrations of T in each treatment group decreased to different extents, and the biggest drop occurred in all treatment groups at 45 days, which was positively correlated with the MT concentration. The 11-KT increased to varying degrees. The results show that: 100 mg/kg was more suitable concentration to bring about sex reversal, MT can cause the early female fish serum E_2 to increase, and exert the effect of androgen, thereby promoting ovarian egg cell degradation, and spermatogonia proliferation.
引文
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[2]姜鹏,韩林强,白俊杰,等.草鱼生长性状的遗传参数和育种值估计[J].中国水产科学,2018,25(1):18-25.Jiang P,Han L Q,Bai J J,et al.Estimation of genetic parameters and breeding values for growth traits in grass carp(Ctenopharyngodon idellus)[J].Journal of Fishery Sciences of China,2018,25(1):18-25(in Chinese).
[3]Shelton W L.Broodstock development for monosex production of grass carp[J].Aquaculture,1986,57(1-4):311-319.
[4]楼允东.鱼类性别控制研究的进展[J].上海水产大学学报,1992,1(3-4):168-173.Lou Y D.Progress on research of sex control in fish[J].Journal of Shanghai Fisheries University,1992,1(3-4):168-173(in Chinese).
[5]方永强,李正森.17α-甲基睾酮对淡水和海水鲻鱼精子发生的比较[J].台湾海峡,1992,11(4):301-304.Fang Y Q,Li Z S.Comparison of 17α-methyltestosterone on spermatogenesis in grey mullet of fresh water and seawater[J].Journal of Oceanography in Taiwan Strait,1992,11(4):301-304(in Chinese).
[6]Yamamoto T O.3 sex differentiation[J].Fish Physiology,1969,3:117-175.
[7]Kitano T,Takamune K,Nagahama Y,et al.Aromatase inhibitor and 17α-methyltestosterone cause sex-reversal from genetical females to phenotypic males and suppression of P450 aromatase gene expression in Japanese flounder(Paralichthys olivaceus)[J].Molecular Reproduction&Development,2000,56(1):1-5.
[8]Blázquez M,Piferrer F,Zanuy S,et al.Development of sex control techniques for European sea bass(Dicentrarchus labrax L.)aquaculture:effects of dietary 17α-methyltestosterone prior to sex differentiation[J].Aquaculture,1995,135(4):329-342.
[9]El-Greisy Z A,El-Gamal A E.Monosex production of tilapia,Oreochromis niloticus using different doses of17α-methyltestosterone with respect to the degree of sex stability after one year of treatment[J].The Egyptian Journal of Aquatic Research,2012,38(1):59-66.
[10]Guerrero R D I.Culture of male tilapia O.mossambica produced through artificial sex reversal[J].Advances in Aquaculture,1976.
[11]Macintosh D J,Varghese T J,Satyanarayana G P R.Hormonal sex reversal of wild-spawned tilapia in India[J].Journal of Fish Biology,1985,26(2):87-94.
[12]郑曙明,陈章宝,贺蓉.甲基睾酮对孔雀鱼、玛丽鱼性逆转的试验[J].四川动物,1997,16(4):183-185.Zheng S M,Chen Z B,He R.The sex revesal of methyltestosterone against Guppies and Mariposities[J].Sichuan Journal of Zoology,1997,16(4):183-185(in Chinese).
[13]Pandian T J,Sheela S G.Hormonal induction of sex reversal in fish[J].Aquaculture,1995,138(1-4):1-22.
[14]Zheng Y,Chen J Z,Liu Y,et al.Molecular mechanism of endocrine system impairment by 17α-methyltestosterone in gynogenic Pengze crucian carp offspring[J].Ecotoxicology and Environmental Safety,2016,128:143-152.
[15]Devlin R H,Nagahama Y.Sex determination and sex differentiation in fish:an overview of genetic,physiological,and environmental influences[J].Aquaculture,2002,208(3-4):191-364.
[16]Wang Y P,Lu Y,Zhang Y,et al.The draft genome of the grass carp(Ctenopharyngodon idellus)provides insights into its evolution and vegetarian adaptation[J].Nature Genetics,2015,47(6):625-631.
[17]刘筠,刘素孋,寿孝钟,等.草鱼性腺发育的研究[J].湖南师范大学自然科学学报,1962(4):65-87,89-98.Liu J,Liu S L,Shou X Z,et al.The research of grass carp gonad development[J].Journal of Natural Science of Hunan Normal University,1962(4):65-87,89-98(in Chinese).
[18]徐永江,柳学周,王清印,等.半滑舌鳎(Cynoglossus semilaevis)血浆性类固醇激素表达与卵巢发育及温光调控的关系研究[J].海洋与湖沼,2011,42(1):67-74.Xu Y J,Liu X Z,Wang Q Y,et al.Relationships between serum sex steriods levels and gonadal development and photothermal regulation during the annual maturation of captive Cynoglossus semilaevis Günther[J].Oceanologia et Limnologia Sinica,2011,42(1):67-74(in Chinese).
[19]洪万树,吴秋艳,张其永.中华乌塘鳢血清性类固醇激素含量与性腺发育的关系及其季节变化[J].厦门大学学报(自然科学版),2009,48(2):274-277.Hong W S,Wu Q Y,Zhang Q Y.Changes of serum sex steroids levels with gonadal development and seasons in Bostrichthys sinensis[J].Journal of Xiamen University(Natural Science Edition),2009,48(2):274-277(in Chinese).
[20]张立涛.斑马鱼早期性腺发育及甲基睾酮对其性分化的影响[D].保定:河北大学,2010.Zhang L T.Development of gonad at early stages in zebrafish(Danio rerio)and effects of MT on its sex differentiation[D].Baoding:Hebei University,2010(in Chinese).
[21]李广丽,刘晓春,林浩然.17α-甲基睾酮对赤点石斑鱼性逆转的影响[J].水产学报,2006,30(2):145-150.Li G L,Liu X C,Lin H R.Effects of 17α-methyltestosterone on sex reversal in red-spotted grouper,Epinephelus akaara[J].Journal of Fisheries of China,2006,30(2):145-150(in Chinese).
[22]Fenske M,Segner H.Aromatase modulation alters gonadal differentiation in developing zebrafish(Danio rerio)[J].Aquatic Toxicology,2004,67(2):105-126.
[23]Rinchard J,Dabrowski K,Garcia-Abiado M A,et al.Uptake and depletion of plasma 17α-methyltestosterone during induction of masculinization in muskellunge,Esox masquinongy:effect on plasma steroids and sex reversal[J].Steroids,1999,64(8):518-525.
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