IGF2BP1 stabilizes Akt2 mRNA to promote glucose metabolism and maintain spermatogonial proliferation.

IF 3.7 3区生物学 Q1 DEVELOPMENTAL BIOLOGY

Reproduction Pub Date : 2025-03-03 Print Date: 2025-04-01 DOI:10.1530/REP-24-0202

Yanan Liu, Yuanyuan Chen, Jiahui Fan, Ziqian Min, Xiaowen Liu, Zenghui Mao, Anji Chen, Min Liu, Rong Xia, Lifang Yang, Dan Li

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引用次数: 0

Abstract

In brief: The dysregulation of spermatogonial proliferation is one of the causes of male infertility. Using mice as an animal model, this study reveals the function and mechanism of m6A methylation reader insulin-like growth factor-2 mRNA-binding protein 1 (IGF2BP1) and its target Akt2 in spermatogonia.

Abstract: IGF2BP1, as a newly identified N6-methyladenosine (m6A) methylation reader, plays a key role in mRNA stability, alternative splicing and translation. However, the function of IGF2BP1 and its target genes in the development of male germ cells remains unclear. In the present study, Western blotting, immunofluorescence and other cellular methods were used to confirm that IGF2BP1 is expressed throughout the male germline with high abundance in spermatogonia of mice, and that its downregulation inhibits spermatogonia proliferation in vitro. Through RNA sequencing, dual luciferase reporter assays, RIP-qPCR and mRNA stability experiments, it was identified that Akt2 is one of the target genes of IGF2BP1. By stabilizing Akt2 mRNA in an m6A-dependent manner, IGF2BP1 maintains spermatogonial proliferation. Furthermore, co-immunoprecipitation and mass spectrometry analyses revealed that PABPC1, DDX5 and FXR1 interact with IGF2BP1, and that the interaction between IGF2BP1 and PABPC1 promotes AKT2 expression. Finally, following the knockdown of both IGF2BP1 and AKT2, glucose and ATP levels in C18-4 and GC-1 spermatogonia cells were found to be reduced, indicating that IGF2BP1 regulates glucose metabolism homeostasis by stabilizing Akt2 expression, thereby influencing spermatogonia proliferation. In addition, analysis of the cryptorchidism database from NCBI revealed that the expression of IGF2BP1 and AKT2 is significantly downregulated in cryptorchid patients with oligospermia or azoospermia. In conclusion, our study elucidates the role and underlying mechanism of IGF2BP1 in spermatogonia, providing a candidate target for male infertility.

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IGF2BP1稳定Akt2 mRNA，促进葡萄糖代谢，维持精原细胞增殖。

胰岛素样生长因子2 mRNA结合蛋白1 （IGF2BP1）是新发现的n6 -甲基腺苷（m6A）甲基化解读器，在mRNA稳定性、选择性剪接和翻译中起关键作用。然而，IGF2BP1及其靶基因在男性生殖细胞发育中的作用尚不清楚。本研究通过Western blotting、免疫荧光等细胞方法证实了IGF2BP1是否在小鼠精原细胞中全雄性种系高丰度表达，以及其下调是否在体外抑制精原细胞增殖。通过RNA测序、双荧光素酶报告基因测定、RIP-qPCR和mRNA稳定性实验，确定Akt2是IGF2BP1的靶基因之一。IGF2BP1通过m6a依赖的方式稳定Akt2 mRNA，维持精原细胞增殖。此外，共免疫沉淀和质谱分析显示，PABPC1、DDX5和FXR1与IGF2BP1相互作用，IGF2BP1和PABPC1相互作用促进AKT2的表达。最后，IGF2BP1和AKT2同时下调后，发现C18-4和GC-1精原细胞中葡萄糖和ATP水平降低，表明IGF2BP1通过稳定AKT2表达调节糖代谢稳态，从而影响精原细胞增殖。此外，NCBI对隐睾数据库的分析显示，IGF2BP1和AKT2在少精子症或无精子症的隐睾患者中表达显著下调。总之，我们的研究阐明了IGF2BP1在精原细胞中的作用及其潜在机制，为男性不育提供了一个候选靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Reproduction 生物-发育生物学

CiteScore

7.40

自引率

2.60%

发文量

199

审稿时长

4-8 weeks

期刊介绍： Reproduction is the official journal of the Society of Reproduction and Fertility (SRF). It was formed in 2001 when the Society merged its two journals, the Journal of Reproduction and Fertility and Reviews of Reproduction. Reproduction publishes original research articles and topical reviews on the subject of reproductive and developmental biology, and reproductive medicine. The journal will consider publication of high-quality meta-analyses; these should be submitted to the research papers category. The journal considers studies in humans and all animal species, and will publish clinical studies if they advance our understanding of the underlying causes and/or mechanisms of disease. Scientific excellence and broad interest to our readership are the most important criteria during the peer review process. The journal publishes articles that make a clear advance in the field, whether of mechanistic, descriptive or technical focus. Articles that substantiate new or controversial reports are welcomed if they are noteworthy and advance the field. Topics include, but are not limited to, reproductive immunology, reproductive toxicology, stem cells, environmental effects on reproductive potential and health (eg obesity), extracellular vesicles, fertility preservation and epigenetic effects on reproductive and developmental processes.