Abstract

Background: Mural Granulosa Cells (MGCs) and Cumulus Cells (CCs) are two specialized cell types that differentiate from a common progenitor during folliculogenesis. Although these two cell types have specialized functions and gene expression profiles, little is known about their microRNA (miRNA) expression patterns.Objective: To describe the miRNA profile of mural and cumulus granulosa cells from human pre-ovulatory follicles.Methods: Using small RNA sequencing, we defined the miRNA expression profiles of human primary MGCs and CCs, isolated from healthy women undergoing ovum pick-up for in vitro Fertilization (IVF).Results: Small RNA sequencing revealed the expression of several hundreds of miRNAs in MGCs and CCs with 53 miRNAs being significantly differentially expressed between MGCs and CCs. We validat-ed the differential expression of miR-146a-5p, miR-149-5p, miR-509-3p and miR-182-5p by RT-qPCR. Analysis of proven targets revealed 37 targets for miR-146a-5p, 43 for miR-182-5p, 2 for miR-509-3p and 9 for miR-149-5p. Gene Ontology (GO) analysis for these 4 target gene sets revealed enrichment of 12 GO terms for miR-146a-5p and 10 for miR-182-5p. The GO term ubiquitin-like protein conjugation was enriched within both miRNA target gene sets.Conclusion: We generated miRNA expression profiles for MGCs and CCs and identified several dif-ferentially expressed miRNAs.

Highlights

  • Folliculogenesis is a complex and precisely regulated process essential for human reproduction

  • Small RNA sequencing revealed the expression of several hundreds of miRNAs in Mural Granulosa Cells (MGCs) and Cumulus Cells (CCs) with 53 miRNAs being significantly differentially expressed between MGCs and CCs

  • We validated the differential expression of miR-146a-5p, miR-149-5p, miR-509-3p and miR-182-5p by Reverse Transcriptase (RT)-qPCR

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Summary

Introduction

Folliculogenesis is a complex and precisely regulated process essential for human reproduction. During growth and maturation of the ovarian follicle, granulosa cells divide and differentiate into a multilayer of Mural Granulosa Cells (MGCs) that line the follicular compartment and a cluster of Cumulus Cells (CCs) that tightly surround the oocyte [1]. Even though these two cell types have a common progenitor in the primordial follicle, their differentiation during folliculogenesis results in cells with specific functions and gene. Mural Granulosa Cells (MGCs) and Cumulus Cells (CCs) are two specialized cell types that differentiate from a common progenitor during folliculogenesis These two cell types have specialized functions and gene expression profiles, little is known about their microRNA (miRNA) expression patterns

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