再生水景观利用对斑马鱼(Danio rerio)下丘脑-垂体-性腺(HPG)轴相关基因转录水平的影响
Effects of Reclaimed Water in Landscape Utilization on Gene Transcription of Hypothalamus-Pituitary-Gonadal (HPG) Axis in Zebrafish (Danio rerio)
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摘要: 再生水对水体生物具有潜在的危害。为了评估再生水对水体生物的早期分子毒理效应，采用荧光定量多聚酶链式反应(polymerase chain reaction, PCR)技术分析再生水对斑马鱼下丘脑-垂体-性腺(hypothalamus-pituitary-gonadal, HPG)轴相关基因转录水平的影响。再生水暴露后，斑马鱼体长、体质量、生长状况因子(condition factor, K)、脑指数(brain-somatic index, BSI)和性腺指数(gonadosomatic index, GSI)无显著改变。再生水暴露7 d引起雄性斑马鱼脑雄激素受体(androgen receptor, Ar)、雌激素受体1 (estrogen receptor 1, Esr1)、卵泡刺激素β多肽(follicle stimulating hormone beta polypeptide, Fshb)、黄体生成素β多肽(luteinizing hormone beta polypeptide, Lhb)、促性腺激素释放激素(gonadotropin-releasing hormone 2, Gnrh2)和促性腺激素释放激素(gonadotropin-releasing hormone 3, Gnrh3)基因的转录水平显著下调(P<0.05)，并引起雌性斑马鱼脑Gnrh3基因转录水平显著上调(P<0.05)。再生水暴露7 d导致类固醇激素合成通路下游基因如Ar、Esr1、卵巢芳香化酶(ovarian cytochrome P450 aromatase, cyp19ala)、3β-羟基类固醇脱氢酶(3β-hydroxysteroid dehydrogenase, hsd3b)、17β-羟化类固醇脱氢酶3(17β-hydroxysteroid dehydrogenase type 3, hsd17b3)和20β-羟化类固醇脱氢酶(20β-hydroxysteroid dehydrogenase, hsd20b)在雄性斑马鱼睾丸中表达水平显著减少(P<0.05)；相反，雌性斑马鱼卵巢Esr1和11β-羟化酶(11β-hydroxylase, cyp11b)基因转录水平显著增加(P<0.05)。再生水对斑马鱼暴露14 d后，雄性斑马鱼脑Esr1、Fshb和Lhb基因表达水平显著下调(P<0.05)，睾丸Ar、Esr1、类固醇源性急性调节蛋白(steroidogenic acute regulatory, star)、cyp19a1a、11β-羟化类固醇脱氢酶2(11β-hydroxysteroid dehydrogenase 2, hsd11b2)、hsd20b和hsd17b3基因转录水平也显著降低(P<0.05)。然而再生水暴露14 d显著上调了雌性斑马鱼卵巢Esr1、孕激素受体(progesterone receptor, pgr)、cyp19a1a、hsd3b、hsd17b3和star基因转录水平(P<0.05)，并且下调了脑Gnrh3和Fshb的转录水平(P<0.05)。综上所述，本研究结果表明再生水显著影响了斑马鱼HPG轴相关基因转录水平，具有性别和暴露时间差异性，为再生水对斑马鱼生殖系统的影响提供了新的见解。
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关键词:
- 再生水 /
- 斑马鱼(Danio rerio) /
- HPG轴 /
- 内分泌干扰效应
Abstract: Reclaimed water (RW) can be potentially harmful to aquatic organisms. To evaluate early molecular responses of reclaimed water on aquatic organisms, this study investigated the impact of RW on gene transcription of the hypothalamus-pituitary-gonadal (HPG) axis of adult zebrafish (Danio rerio) using the fluorescence quantitative polymerase chain reaction (PCR). After exposure, the total body length, wet weight, condition factor (K), brain-somatic index (BSI) and gonadosomatic index (GSI) did not change significantly. In brain, RW exposure for 7 d caused significant down-regulation in the transcripts of androgen receptor (Ar), estrogen receptor 1 (Esr1), follicle stimulating hormone beta polypeptide (Fshb), luteinizing hormone beta polypeptide (Lhb), gonadotropin-releasing hormone 2 (Gnrh2) and gonadotropin-releasing hormone 3 (Gnrh3) genes in male (P<0.05) and up-regulation in the transcript of Gnrh3 gene in female zebrafish (P<0.05). After exposure to RW for 7 d, the downstream genes of steroidogenic pathway like Ar, Esr1, ovarian cytochrome P450 aromatase (cyp19ala), 3β-hydroxysteroid dehydrogenase (hsd3b), 17β-hydroxysteroid dehydrogenase type 3 (hsd17b3) and 20β-hydroxysteroid dehydrogenase (hsd20b) expression were significantly reduced in testis of male zebrafish (P<0.05); on the contrary, the transcript of Esr1 and 11β-hydroxylase (cyp11b) genes were enhanced in ovary of female zebrafish (P<0.05). After exposure to RW for 14 d, it was found that the expression of Esr1, Fshb and Lhb genes was significantly down-regulated in brain of male zebrafish (P<0.05), and the expression of Ar, Esr1, steroidogenic acute regulatory (star), cyp19a1a, 11β-hydroxysteroid dehydrogenase 2 (hsd11b2), hsd20b and hsd17b3 genes was also significantly reduced in testis of male zebrafish (P<0.05). While in females, RW exposure for 14 d up-regulated the expression of Esr1, progesterone receptor (pgr), cyp19a1a, hsd3b, hsd17b3 and star in ovary (P<0.05), and meanwhile Gnrh3 and Fshb were down-regulated in brain (P<0.05). Overall, these findings exhibited RW have significant effects on the transcript of genes of HPG axis, showing sex and exposure time specific effects, which provide new insights of RW effects on zebrafish reproductive system. -
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