Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, NMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology Products, Co-Innovation Center of Neuroregeneration, Nantong University, Nantong 226001, China.
Department of Pharmacology, School of Pharmacy, Nantong University, Nantong 226001, China.
Nutrients. 2021 Dec 24;14(1):66. doi: 10.3390/nu14010066.
Use of folic acid (FA) during early pregnancy protects against birth defects. However, excess FA has shown gender-specific neurodevelopmental toxicity. Previously, we fed the mice with 2.5 times the recommended amount of FA one week prior to mating and during the pregnancy and lactation periods, and detected the activated expression of and related genes in the brains of weaning male offspring, as well as behavioral abnormalities in the adults. Here, we studied whether female offspring were affected by the same dosage of FA. An open field test, three-chamber social approach and social novelty test, an elevated plus-maze, rotarod test and the Morris water maze task were used to evaluate their behaviors. RNA sequencing was performed to identify differentially expressed genes in the brains. Quantitative real time-PCR (qRT-PCR) and Western blots were applied to verify the changes in gene expression. We found increased anxiety and impaired exploratory behavior, motor coordination and spatial memory in FA-exposed females. The brain transcriptome revealed 36 up-regulated and 79 down-regulated genes in their brains at weaning. The increase of ; ; ; ; ; and , as well as a decrease of mRNA were confirmed by qRT-PCR. Among these genes; the mRNA levels of were increased in the both FA-exposed female and male brains. The elevation of Sult1a1 protein was confirmed by Western blots. Our data suggest that excess FA alteres brain gene expression and behaviors in female offspring, of which certain genes show apparent gender specificity.
在怀孕早期使用叶酸（FA）可以预防出生缺陷。然而，过量的 FA 已显示出具有性别特异性的神经发育毒性。此前，我们在交配前一周和怀孕期间以及哺乳期给老鼠喂食推荐剂量的 2.5 倍 FA，检测到断奶雄性后代大脑中 和相关基因的激活表达，以及成年动物的行为异常。在这里，我们研究了相同剂量的 FA 是否会影响雌性后代。通过旷场试验、三箱社交接近和社交新颖性试验、高架十字迷宫、转棒试验和 Morris 水迷宫任务来评估它们的行为。进行 RNA 测序以鉴定大脑中差异表达的基因。应用定量实时 PCR（qRT-PCR）和 Western blot 验证基因表达的变化。我们发现暴露于 FA 的雌性动物表现出焦虑增加和探索行为、运动协调和空间记忆受损。大脑转录组在断奶时显示出 36 个上调和 79 个下调的基因。qRT-PCR 证实了 、 、 、 、 、 和 的增加； 以及 的 mRNA 减少。在这些基因中；FA 暴露的雌性和雄性大脑中的 的 mRNA 水平均升高。Western blot 证实了 Sult1a1 蛋白的升高。我们的数据表明，过量的 FA 改变了雌性后代的大脑基因表达和行为，其中某些基因表现出明显的性别特异性。
