Paternal fenvalerate exposure causes depressive-like behaviour by altering Grb10 gene DNA methylation in adolescent offspring

父代氰戊菊酯暴露通过改变印记基因GRB10 DNA甲基化水平导致青春期子代抑郁样行为

Authors：Hui-Ru Chen, Tao Wang, Jing Shao, Hui-Min Zhu, Chi-Zhang, Ting Zhao, Li-Hua Xu, Min Wang, Jing-Jing Li, Qi-Long Zhu, Xi-Meng Qi, De-Xiang Xu, Bo Wang, Xiu-Hong Meng

Source：Ecotoxicology And Environmental Safety 

DOI: 10.1016/j.ecoenv.2025.117994

Abstract

Fenvalerate, a typical pyrethroid pesticide, is a neurological toxicant. This study aimed to evaluate the effect of paternal exposure to fenvalerate on depressive-like behaviours in adolescent offspring. Depression-like behavior was determined by Sucrose Preference Test (SPT), Tail Suspension Test (TST) and Forced Swimming Test (FST) in adolescent offspring. The level of dopamine was reduced in the midbrain of fenvalerate-exposed adolescent offspring. Tyrosine hydroxylase (Th), a rate limiting enzyme for dopamine synthesis, was significantly reduced in the midbrain of adolescent offspring exposed to fenvalerate. And Th was decreased in the midbrain and hindbrain of fetuses exposed to fenvalerate. Transcriptome analysis revealed growth factor receptor-bound protein 10 (Grb10) was decreased in the fetal hindbrain exposed to fenvalerate. Grb10 mRNA and protein were reduced in the fetal hindbrain exposed to fenvalerate. Interestingly, in vitro experiments, Th was reduced by si-Grb10. Conversely, Th was increased by oe-Grb10. Mechanistically, the 5mC content of Grb10 gene at one CpG fragment was reduced in the fetal hindbrain exposed to fenvalerate. And the 5mC content of Grb10 gene at eighteen CpG sites was decreased in paternal sperm exposed to fenvalerate. In summary, paternal fenvalerate exposure causes depressive-like behavior by altering DNA methylation of Grb10 gene in the sperm. 

Keywords: DNA methylation; Depression-like behaviour; Dopamine; Fenvalerate; Grb10; Paternal; Th.

摘要：

背景：近年来，抑郁症成为全球最普遍的情绪障碍疾病，其中环境暴露是导致抑郁症的主要原因之一。研究表明孕期氰戊菊酯暴露可导致子代抑郁样行为。然而，父代氰戊菊酯暴露对青春期子代抑郁样行为的影响及其机制尚不清楚。

结果：在抑郁症行为学实验中，糖水偏好实验结果显示氰戊菊酯暴露组的蔗糖溶液消耗量明显低于对照组，这提示暴露组小鼠快感缺失，是抑郁样行为形成的原因之一。尾部悬挂实验和强迫游泳实验结果显示，氰戊菊酯暴露组的不动时间明显高于对照组，这提示暴露组小鼠出现绝望和消极状态，这也是抑郁样行为形成的原因之一。酶联免疫吸附技术显示，父代氰戊菊酯暴露降低多巴胺含量。免疫组织化学实验结果表明，父代氰戊菊酯暴露可减少青春期子代VTA TH阳性细胞面积。通过RT-PCR和Western Blot实验检测青春期子代VTA多巴胺合成限速酶TH的表达水平，结果发现青春期子代VTA TH mRNA和蛋白表达水平下降。此外，RT-PCR和Western blot实验发现父代氰戊菊酯暴露降低胎鼠中脑TH mRNA和蛋白表达水平。转录组RNA-seq显示，按P≤0.05水平检测到胎鼠后脑GRB10 mRNA表达水平显著下降。通过RT-PCR实验发现父代氰戊菊酯暴露降低胎鼠后脑GRB10 mRNA表达水平。最后，细胞实验结果显示，GRB10 siRNA的降低MN9D细胞中TH表达水平。胎鼠后脑的DNA甲基化测序显示，与对照组相比，父代氰戊菊酯暴露的胎鼠后脑中GRB10_10 CpG片段的1个CpG位点（11974988）的5mC含量降低，差异有统计学意义。父代精子DNA甲基化测序表明，与对照组相比，父代氰戊菊酯暴露的父代精子中GRB10_7、GRB10_8、GRB10_9、GRB10_10和GRB10_11 CpG片段的5mC含量降低。随后，我们进一步检测GRB10CpG片段中富含5mC的CpG位点，研究表明，父代氰戊菊酯暴露的父代精子11个CpG位点（12037217、11973406、11973437、11973507、11973501、11973543、11973606、11973767、11973883、11974988和11975147）的5mC含量降低，差异均有统计学意义。

结论：父代氰戊菊酯暴露可能通过干扰GRB10 DNA甲基化水平导致多巴胺含量下降，从而诱发青春期子代小鼠抑郁样行为。 

关键词：父代 氰戊菊酯 抑郁样行为 多巴胺 GRB10 DNA甲基化