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01.12.2014 | Research | Ausgabe 1/2014 Open Access

Reproductive Biology and Endocrinology 1/2014

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

Zeitschrift:
Reproductive Biology and Endocrinology > Ausgabe 1/2014
Autoren:
Jingying Hu, Shaoyang Sun, Meng Guo, Houyan Song
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1477-7827-12-40) contains supplementary material, which is available to authorized users.

Competing interests

The authors declare that they have no competing interests.

Authors’ contributions

JYH participated in the design of the study, and drafted the manuscript. JYH and SYS conducted the data collection. JYH and MG participated in the statistical analysis, and performed the diagramming. HYS conceived of the study, participated in its design and coordination, and helped to draft the manuscript. All the authors read and approved the final manuscript.

Abstract

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.
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