性类固醇激素对胡子鲇性分化的影响
EFFECTS OF SEX STEROID HORMONES ON SEX DIFFERENTIATION OF CLARIAS FUSCUS
-
摘要: 研究采用组织学和荧光实时定量PCR方法，检测17-甲基睾酮（17-MT）和17-雌二醇（17-E2）对胡子鲇（Clarias fuscus）性腺组织学、性别比率以及性分化前后脑型芳香化酶基因（Cyp19a1b）和翼状螺旋/叉头转录因子2（Foxl2）基因表达的影响。结果表明：在出膜后230日龄，17-MT （50、100和200 g/L）浸浴、17-E2（100、200和300 g/g）投喂处理对成活率无显著影响，但中、高剂量（100和200 g/L）17-MT显著抑制卵巢发育，促进精巢发育，卵巢腔出现时间分别推迟4d和6d，初级卵母细胞出现时间分别推迟8d和9d，而初级精母细胞出现时间则分别提前3d和5d，且雄性率分别达70%和76%，显著高于50 g/L组和对照组（P0.05）。相反，中、高剂量17-E2（200和300 g/g）处理使卵巢腔出现时间分别提前2d和3d，初级卵母细胞出现时间分别提前1d和3d，初级精母细胞出现时间分别推迟3d和7d，而雌性率分别达74%和78%，显著高于100 g/g组和对照组（P0.05）。此外，在性腺分化期，17a-MT促进Cyp19a1b但抑制Foxl2的表达，而17-E2促进Cyp19a1b和 Foxl2的表达。结果显示Cyp19a1b不是引起胡子鲇性分化的直接因素，但可能通过下丘脑-垂体-性腺轴对胡子鲇性分化过程产生间接影响；而Foxl2直接参与胡子鲇的性分化，即17a-MT和17-E2分别通过抑制和促进Foxl2的表达来影响雌激素的生物合成，从而调控胡子鲇性分化的方向。Abstract: Clarias fuscus, a common freshwater fish in China, was selected as our experiment material. Two-day juvenile C. fuscus was divided into two groups. They were immersed in different doses of 17-methyltestosterone (50, 100, and 200 g/L 17-MT) or fed with 17-estradiol (100, 200, and 300 g/g 17-E2) for 30 days. Effects of 17-MT and 17-E2 on survival rate, sex ratio, gonad histology, and Foxl2 and Cyp19a1b expressions were examined by morphologic observation, histology and Real time fluorescent quantitative PCR during its period of sex differentiation (230d after hatching). The results showed that 17-MT and 17-E2 had no influence on survival rate but affected sex ratio and the time of gonadal differentiation. Doses of 17-MT at 100 and 200 g/L produced more males (70% and 76%, respectively) than 50 g/L 17-MT (54%) did (P0.05), but no significant difference in sex ratio was observed between the 50 g/L 17-MT treated group and the control group (56%). In addition, the former accelerated the occurrence of primary spermocytes for three and five days, but deferred that of ovarian cavity for four and six days, and primary oocytes for eight and nine days, respectively. In contrast, doses of 17-E2 at 200 and 300 g/g produced more males (74% and 78%, respectively) than the control group (P0.05), but no significant difference in sex ratio was observed between the 100 g/g 17-E2 treated group (66%) and the control group (56%). Dose of 17-E2 at 200 and 300 g/g accelerated the occurrence of ovarian cavity for two and three days, and primary oocytes for one and three days, respectively, but deferred that of primary spermatocytes for three and seven days. The expressions of Foxl2 and Cyp19a1b showed that dose of 200 g/L 17-MT increased the expression of Cyp19a1b but inhibited that of Foxl2.However, dose of 300g/g 17-E2 increased the expression both of Cyp19a1band Foxl2.The results suggested that Foxl2 but not Cyp19a1b involved in mediating sex differentiation directly in C. fuscus, and 17-MT inhibited but 17-E2 promoted the expressions of Foxl2 toinfluence the estrogen biosynthesis, which controlled the sex differentiation. However, Cyp19a1b played an indirect role on sex differentiation by acting on the hypothalamic-pituitary-gonadal axis.
-
Keywords:
- Clarias fuscus /
- Sex differentiation /
- 17-MT /
- 17-E2 /
- Cyp19a1b /
- Foxl2
-
-
[1] Ospina-Alvarez N,Piferrer F.Temperature-dependent sex determination in fish revisited: prevalence,a single sex ratio response pattern,and possible effects of climate change[J].Public Library of Science One,2008,3(7): 1-11
[2] Baroiller J F,D'Cotta H.Environment and sex determination in farmed fish[J].Comparative Biochemistry and Physiology C Toxicology and Pharmacology,2001,130(4): 399-409
[3] Lin H R.Fish Physiology (2nd)[M].Guangzhou: Guangdong High Education Press.2007,205-212[林浩然.鱼类生理学(第二版),广州: 广东高等教育出版社.2007,205-212]
[4] Wang D S,Zhou L Y,Kobayashi T.Doublesex-and Mab-3-related transcription factor-1 repression of aromatase transcription,a possible mechanism favoring the male pathway in tilapia[J].Endocrinology,2010,151(3): 1331-1340
[5] Delvin 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
[6] Guiguen Y,Fostier A,Piferrer F,et al.Ovarian aromatase and estrogens: A pivotal role for gonadal sex differentiation and sex change in fish[J].General and Comparative Endocrinology,2010,165(3): 352-366
[7] Park I S,Jung H K,Sung H C.Sex differentiation and hormonal sex reversal in the bagrid catfishPseudobagrus fulvidraco[J].Aquaculture,2004,232(1-4): 183-193
[8] Baron D,Coequet J,Xia X,et al.An evolutionary and functional analysis of Foxl2 in rainbow trout gonad differentiation[J].Journal of Molecular Endocrinology,2004,33: 705-715
[9] Baron D,Houlgatte R,Fostier A,et al.Expression profiling of candidate genes during ovary-to-testis trans-differentiation in rainbow trout masculinized by androgens[J].General and Comparative Endocrinology,2008,156(2): 369-378
[10] Kobayashi T,Kajiura-Kobayashi H,Nagahama Y.Induction of XY sex reversal by estrogen involves altered gene expression in a teleost,tilapia[J].Cytogenetic Genome Research,2003, 101(3-4): 289-294
[11] Wang W D,Zhu C H,Deng S P,et al. Occurrence and differentiation of gonad inClarias fuscus[J].Acta Hydrobiologica Sinica,2012, 36(5): 993-997[王文达,朱春华,邓思平,等.胡子鲇性腺发生与分化组织学研究.水生生物学报,2012,36(5): 993-997]
[12] Sun J,Li G L,Zhu C H,et al.Molecular cloning and expression ofCyp19a1b cDNA inClarias fuscus[J].Journal of Fishery Sciences of China,2012,19(3): 408-415[孙晶,李广丽,朱春华,等.胡子鲇脑型芳香化酶基因全长cDNA克隆及表达.中国水产科学,2012,19(3): 408-415]
[13] Deng S P,Wang J J,Wu T L,et al.cDNA cloning and expression analysis of Dmrt1inClarias fuscus[J].Acta Hydrobiologica Sinica,2012,36(4): 610-617[邓思平,王静杰,吴天利,等.胡子鲇Dmrt1基因全长cDNA克隆及其表达分析.水生生物学报,2012,36(4): 610-617]
[14] Yao D X.Sex differentiation and hormonal sex reversal ofPelteobagrus fuIvidraco[D].Thesis for Master of Science,Northeast Forestry University,Harbin.2007[姚道霞.黄颡鱼性分化及激素诱导性转化研究.硕士学位论文,东北林业大学,哈尔滨.2007]
[15] Zhang X Y,Liu H J.Effects of 17-Estradiol on sex differentiation and growth in half-smooth tongue-sole (Cynoglossus semilaevis)[J].Journal of Northeast Agricultural University,2009,40(6): 67-72[张晓彦,刘海金.17-雌二醇对半滑舌鳎性分化和生长的影响.东北农业大学学报,2009,40(6): 67-72]
[16] Zhuo X L, Zou J X, Cui K,et al.The influence of exogenous methyl testosterone on the gonad developmental process of female and intersexualMonopterus albus[J].Acta Hydrobiologica Sinica,2008,32(6): 861-867[卓孝磊,邹记兴,崔科,等.外源性甲基睾丸酮对雌性和间性黄鳝性腺发育的影响.水生生物学报,2008,32(6): 861-867]
[17] Li G L,Liu X C,Lin H R.Effects of 17a-methyltestosterone on sex reversal in red-spotted grouper,Epinephelus akaara[J].Journal of Fisheries of China,2006,30(2): 145-150[李广丽,刘晓春,林浩然.17-甲基睾酮对赤点石斑鱼性逆转的影响.水产学报,2006,30(2): 145-150]
[18] Nakamura M,Kobayashi T,Chang X T,et al.Gonadal sex differentiation in teleost fish[J].Journal of Experimental Biology,1998,281(5): 362-372
[19] Hendry C I,Martin-Robichaud D J,Benfey T J.Hormonal sex reversal of Atlantic Halibut (Hippoglossus hippoglossus)[J].Aquaculture,2003,219(1-4): 769-782
[20] Chen S L,Deng S P,Ma H Y,et al.Molecular marker- assisted sex control inCynoglossus semilaevis[J].Aquaculture,2008,283(1-4): 7-12
[21] Johnstone R,Simpson T H,Youngson A F.Sex reversal in salmonid culture: PartⅡ.The progeny of sex-reversal rainbow trout[J].Aquaculture,1979,18: 13-19
[22] Piferrer F,Zanuy S,Carrillo M,et al. Brief treatment with an aromatase inhibitor during sex differentiation causes chromosomally female salmon to develop as normal functional males[J].ExperimentalZoology,1994,270(3): 255-262
[23] Bjerselius R,Lundstedt-Enkel K,Olsen H,et al. Male goldfish reproductive behaviour and physiology are severely affected by exogenous exposure to 17-estradiol[J].Aquatic Toxicology,2001,53(2): 139-152
[24] Trant J M,Gavasso S,Ackers J,et al.Developmental expression of cytochrome P450 aromatase genes (CYP19a and CYP19b) in zebrafish fry (Danio rerio)[J].ExperimentalZoology,2001,290(5): 475-483
[25] Halm S,Pounds N,Maddix S,et al.Exposure to exogenous 17-oestradiol disrupts P450aromB mRNA expression in the brain and gonad of adult fathead minnows (Pimephales promelas)[J].Aquatic Toxicology,2002,60(3-4): 285-299
[26] Kishida M,Callard G V.Distinct cytochrome P450 aromatase isoforms in zebrafish (Danio rerio) brain and ovary are differentially programmed and estrogen regulated during early development[J].Endocrinology,2001,142(2): 740-750
[27] Gelinas D,Pitoc G A,Callard G V.Isolation of goldfish brain cytochrome P450 aromatase cDNA: mRNA expression during the seasonal cycle and after steroid treatment[J].Molecular and Cellular Endocrinology,1998,138(1-2): 81-93
[28] Li G L,Liu X C,Lin H R.Effects of aromatizable and nonaromatizable androgens on the sex inversion of red-spotted grouper (Epinephelus akaara)[J].Fish Physiology and Biochemistry,2006,32(1): 25-33
[29] Liley N,Stacey N E.Hormones,Pheromones and Reproductive Behaviour in Fish[M].In: Hoar W S,Randall D J (Eds.),Fish Physiology.New York: Academic Press.1983,1-63
[30] Wu R F.Clone,expression ofFoxl2gene and its function on sex differentiation onSilurus meridionalis[D].Thesis for Master of Science.Southwest University,Chongqing.2007[吴瑞凤.南方鲇Foxl2基因克隆、表达及其在性腺分化中作用的初步研究.硕士学位论文,西南大学,重庆.2007]
-
期刊类型引用(6)
1. 张东云，宋红梅，李胜杰，田太行，杜金星，朱涛，雷彩霞，朱俊杰. 17β-雌二醇和曲洛斯坦对大口黑鲈生长及性腺发育的影响. 中国水产科学. 2024(03): 266-277 . 
百度学术
2. 沈奕君，朱奕安，杨志华，叶明慧，周大颜，李广丽，田昌绪. 胡子鲇qRT-PCR内参基因的筛选. 渔业科学进展. 2024(05): 144-154 . 百度学术
3. 林星桦，周大颜，沈奕君，吴冠佐，吴冠佑，陈宇，张玉蕾，肖珊，邓思平，李广丽，田昌绪. 胡子鲇(Clarias fuscus)雌雄基因组Survey分析. 基因组学与应用生物学. 2023(06): 610-619 . 百度学术
4. 吴燕铎，欧密，高丹丹，陈昆慈，罗青，刘海洋，赵建. 斑鳢叉头框转录因子基因Foxl2的克隆表达及性类固醇激素刺激下的表达响应. 大连海洋大学学报. 2022(01): 49-60 . 百度学术
5. 杨超超，潘鹏涛，谭婷，杨娜，雷忻. 内分泌干扰物对鱼类性腺干扰的分子作用机制. 延安大学学报(自然科学版). 2021(03): 27-32 . 百度学术
6. 刘阳，温海深，李吉方，张美昭，王晓龙，常志成，田源. 不同盐度与雌二醇投喂对花鲈幼鱼生长性能的影响. 海洋科学. 2017(06): 9-17 . 百度学术
其他类型引用(5)
计量
- 文章访问数: 1450
- HTML全文浏览量: 6
- PDF下载量: 1015
- 被引次数: 11
下载: