Synthetic progestins levonorgestrel (LNG) and dydrogesterone (DDG) are frequency detected in surface water. Combined effects of LNG and DDG on gonad differentiation are similar to LNG single exposure in juvenile zebrafish. However, LNG and DDG mixtures have stronger effects on spermatogenesis in testes of adult zebrafish, which show variable at different life stage. Effects of LNG and DDG mixtures on eyes and brain remain unknown. Here we investigated effects of LNG, DDG and their mixtures on eyes and brain. Zebrafish were exposed to LNG, DDG and their mixtures from 2 hpf to 144 dpf. Rhythm and vision related biological processes were enriched in eyes and brain in LNG and DDG treatments, which indicated rhythmic oscillation in eyes and brain. The qPCR data revealed that both LNG and DDG decreased transcription of arntl2 and clocka, while increased transcription of per1a, per1b, rpe65a and tefa in eyes and brain. However, DDG and LNG mixtures had slight effect on transcription of genes related to rhythm and vision. In addition, LNG and DDG reduced the thickness of inner nuclear layer in the eyes. Bliss independent model revealed that LNG and DDG had antagonist effects on transcription and histology in eyes and brain. Moreover, LNG and DDG formed the same hydrogen bonds with green-sensitive opsin-4 and rhodopsin kinase GRK7a. Taken together, LNG and DDG competed with each other for the same binding residues resulting in antagonist effect in their mixtures treatments, and have significant ecological implications to assess combined effects of progestins mixtures on fish in different organs.

在地表水中频繁检测到合成孕激素左炔诺孕酮 (LNG) 和地屈孕酮 (DDG)。LNG 和 DDG 对性腺分化的综合影响类似于幼年斑马鱼的 LNG 单次暴露。然而，LNG和DDG混合物对成年斑马鱼睾丸精子发生的影响更强，在不同的生命阶段表现出差异。LNG 和 DDG 混合物对眼睛和大脑的影响仍然未知。在这里，我们研究了 LNG、DDG 及其混合物对眼睛和大脑的影响。斑马鱼暴露于 2 hpf 至 144 dpf 的 LNG、DDG 及其混合物。在 LNG 和 DDG 治疗中，眼睛和大脑中丰富了节律和视觉相关的生物过程，这表明眼睛和大脑有节律性振荡。qPCR 数据显示 LNG 和 DDG 均降低了arntl2的转录和clocka，同时增加per1a、per1b、rpe65a和tefa的转录在眼睛和大脑中。然而，DDG 和 LNG 混合物对节律和视觉相关基因的转录有轻微影响。此外，LNG 和 DDG 降低了眼睛内核层的厚度。Bliss 独立模型显示 LNG 和 DDG 对眼睛和大脑的转录和组织学具有拮抗作用。此外，LNG和DDG与绿色敏感的opsin-4和视紫红质激酶GRK7a形成了相同的氢键。总之，LNG 和 DDG 相互竞争相同的结合残基，从而在它们的混合物处理中产生拮抗作用，并且对评估孕激素混合物对不同器官鱼类的综合影响具有重要的生态意义。