Use of antagonists and morpholinos in loss-of-function analyses: estrogen receptor ESR2a mediates the effects of 17alpha-ethinylestradiol on primordial germ cell distribution in zebrafish

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• 作者：Jingying Hu (3)
  Shaoyang Sun (3)
  Meng Guo (3)
  Houyan Song (3)
  
  3. Department of Biochemistry and Molecular Biology ; Shanghai Medical School and Key Laboratory of Molecular Medicine ; Ministry of Education ; Fudan University ; Shanghai ; 200032 ; PR China
  
• 关键词：17alpha ; ethinylestradiol ; Primordial germ cells ; Estrogen receptor
• 刊名：Reproductive Biology and Endocrinology
• 出版年：2014
• 出版时间：December 2014
• 年：2014
• 卷：12
• 期：1
• 全文大小：353 KB
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• 刊物主题：Reproductive Medicine; Endocrinology;
• 出版者：BioMed Central
• ISSN：1477-7827

文摘

Background Various chemicals released into the aquatic environment adversely affect the reproductive system of fish, particularly by changing gonad structure and function. 17alpha-ethinylestradiol (EE2) is a potent environmental estrogen that disrupts sexual differentiation and normal reproduction in fish. Previous studies have shown that exposure to endocrine-disrupting chemicals (EDCs) disrupts the migration of primordial germ cells (PGCs) in zebrafish. Methods To investigate the effects of EE2 exposure on PGC migration, zebrafish embryos were injected with gfp-nanos mRNA to label PGCs and subsequently exposed to different concentrations of EE2. Typical estrogen receptor antagonist treatment and morpholino knockdown experiments were used to identify functional estrogen receptors that mediate the effects of EE2. Results The migration of PGCs was disrupted after exposure to high concentrations of EE2 (1 mirog/L). Loss-of-function analyses were performed for estrogen receptor ESR1, ESR2a, and ESR2b, and only loss of ESR2a resulted in a decreased number of ectopic PGCs following exposure to 1 mirog/L EE2. Conclusions EE2 exposure disrupts PGC migration and distribution, and this effect is mediated through the estrogen receptor ESR2a.