产前缺氧（PH）是产科最常见的并发症之一，与抑郁、焦虑、认知障碍等多种神经系统疾病密切相关。我们之前的研究发现Zfp462杂合子（Het）小鼠表现出明显的焦虑样行为。有趣的是，患有PH的子代小鼠在成年后也有类似焦虑的行为，伴随着Zfp462表达减少和miR-377-3p表达增加；然而，确切的监管机制仍不清楚。在本研究中，采用蛋白质印迹、基因敲低、免疫荧光、双荧光素酶报告基因测定、免疫沉淀、miR-377-3p模拟物或抑制剂细胞转染、实时定量PCR和救援测定来检测miR-377-3p的变化。产前缺氧后代大脑中的 377-3p-Zfp462-Pbx1（前 B 细胞白血病同源框 1）通路解释了焦虑样行为的发病机制。我们发现Zfp462缺陷通过泛素化促进Pbx1蛋白降解，并且Zfp462 Het小鼠表现出蛋白激酶B（PKB，也称为Akt）-糖原合成酶激酶-3β（GSK3β）-cAMP反应元件结合蛋白（CREB）途径的下调以及具有焦虑样行为的海马神经发生。此外，PH小鼠表现出miR-377-3p上调、Zfp462/Pbx1-Akt-GSK3β-CREB通路活性下调、海马神经发生减少和焦虑样表型。有趣的是，miR-377-3p 直接靶向 Zfp462 mRNA 的 3'UTR 来调节 Zfp462 表达。重要的是，将 miR-377-3p antagomir 显微注射到 PH 小鼠海马齿状回中，上调 Zfp462/Pbx1-Akt-GSK3β-CREB ​​通路活性，增加海马神经发生，并改善焦虑样行为。总的来说，我们的研究结果证明了 miR-377-3p 通过 Zfp462/Pbx1-Akt-GSK3β-CREB ​​通路在海马神经发生和焦虑样行为的调节中发挥着至关重要的作用。因此，miR-377-3p 可能成为产前缺氧后代焦虑样行为的潜在治疗靶点。© 2023。作者获得 Springer Science Business Media, LLC（Springer Nature 旗下子公司）的独家许可。

Prenatal hypoxia (PH) is one of the most common complications of obstetrics and is closely associated with many neurological disorders such as depression, anxiety, and cognitive impairment. Our previous study found that Zfp462 heterozygous (Het) mice exhibit significant anxiety-like behavior. Interestingly, offspring mice with PH also have anxiety-like behaviors in adulthood, accompanied by reduced expression of Zfp462 and increased expression of miR-377-3p; however, the exact regulatory mechanisms remain unclear. In this study, western blotting, gene knockdown, immunofluorescence, dual-luciferase reporter assay, immunoprecipitation, cell transfection with miR-377-3p mimics or inhibitors, quantitative real-time PCR, and rescue assay were used to detect changes in the miR-377-3p-Zfp462-Pbx1 (pre-B-cell leukemia homeobox1) pathway in the brains of prenatal hypoxic offspring to explain the pathogenesis of anxiety-like behaviors. We found that Zfp462 deficiency promoted Pbx1 protein degradation through ubiquitination and that Zfp462 Het mice showed downregulation of the protein kinase B (PKB, also called Akt)-glycogen synthase kinase-3β (GSK3β)-cAMP response element-binding protein (CREB) pathway and hippocampal neurogenesis with anxiety-like behavior. In addition, PH mice exhibited upregulation of miR-377-3p, downregulation of Zfp462/Pbx1-Akt-GSK3β-CREB pathway activity, reduced hippocampal neurogenesis, and an anxiety-like phenotype. Intriguingly, miR-377-3p directly targets the 3'UTR of Zfp462 mRNA to regulate Zfp462 expression. Importantly, microinjection of miR-377-3p antagomir into the hippocampal dentate gyrus of PH mice upregulated Zfp462/Pbx1-Akt-GSK3β-CREB pathway activity, increased hippocampal neurogenesis, and improved anxiety-like behaviors. Collectively, our findings demonstrated a crucial role for miR-377-3p in the regulation of hippocampal neurogenesis and anxiety-like behaviors via the Zfp462/Pbx1-Akt-GSK3β-CREB pathway. Therefore, miR-377-3p could be a potential therapeutic target for anxiety-like behavior in prenatal hypoxic offspring.© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.