Circular RNA expression profiles of mouse ovaries during postnatal development and the function of circular RNA epidermal growth factor receptor in granulosa cells

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Identification of Circular RNAs using RNA Sequencing

Identification of Circular RNAs using RNA Sequencing

Published on: November 14, 2019

Wenchao Jia¹, Bo Xu¹, Ji Wu²

¹Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, Shanghai Jiao Tong University, Shanghai 200240, China.

Metabolism: Clinical and Experimental

|April 11, 2018

Abstract

Purpose:

Circular RNAs (circRNAs) are a class of noncoding RNAs that can regulate gene expression at the post-transcriptional level. The contribution of circRNAs in the regulation of granulosa cells (GCs) functions is not yet clear. The aim of this study was to analyze circRNA expression in adult and neonate ovaries, uncover the biological roles of circ_0002861 (circEGFR) and identify the mechanism by which it modulates follicular development.

Basic Procedures:

The circRNA expression profiles of adult and neonatal mouse ovaries were explored by high-throughput sequencing. The function of circEGFR was measured by RNA fluorescence in situ hybridization, overexpression, knockdown, RNA immunoprecipitation and luciferase reporter assays in GCs.

Main Findings:

Numerous differentially expressed circRNAs were identified in adult and neonatal ovaries. Through circRNAs expression patterns and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, estrogen signaling was found to be upregulated in adult ovaries compared with neonate ovaries. Further analysis revealed that the expression of circEGFR (circ_0002861, ID: mmu_circ_0002861 in circBase) was increased in adult ovaries compared with neonate ovaries. circEGFR is formed by splicing from exons 14 and 15 of the epidermal growth factor receptor (EGFR) gene to produce a covalently linked 249-nucleotide circRNA. Overexpression of circEGFR increased estradiol (E₂) production and GCs growth, whereas circEGFR knockdown enhanced progesterone production and inhibited (E₂) secretion by GCs. Bioinformatic screening identified several binding sites for miR-125a-3p in the circEGFR sequence. RNA immunoprecipitation and luciferase reporter assays demonstrated that circEGFR may act as a sponge for miR-125a-3p, thus modulating Fyn expression.

Principal Conclusions:

These findings illustrate that circEGFR may play a vital role in ovarian GCs by modulating Fyn via competitive binding with miR-125a-3p. Our results suggest potential applications of circEGFR in reproductive and steroid-related disorder therapy.

Keywords:

Circular RNA (circRNA)Competing endogenous RNAs (ceRNA)Estradiol Granulosa cells RNA-Seq

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