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. 2019;8(1):61-67.
doi: 10.2174/2211536607666180912152618.

Differential miRNA Expression Profiles in Cumulus and Mural Granulosa Cells from Human Pre-ovulatory Follicles

Daniela Andrei  1   2 , Roland A Nagy  1   2 , Aafke van Montfoort  1 , Uwe Tietge  2 , Martijn Terpstra  3 , Klaas Kok  3 , Anke van den Berg  4 , Annemieke Hoek  1 , Joost Kluiver  4 , Rogier Donker  1
Affiliations

Differential miRNA Expression Profiles in Cumulus and Mural Granulosa Cells from Human Pre-ovulatory Follicles

Daniela Andrei et al. Microrna. 2019.

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 preovulatory 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 validated 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 differentially expressed miRNAs.

Keywords: Cumulus cells; folliculogenesis; gene ontology; granulosa cells; microRNA; small RNA-seq..

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Figures

Fig. (1)
Fig. (1)
Pie chart showing the relative abundance of the top 10 most abundant miRNAs in mural (MGCs) and cumulus (CCs) granulosa cells as identified by small RNA sequencing analysis. n=3 individual patients.
Fig. (2)
Fig. (2)
Heatmap showing unsupervised clustering of 53 differentially expressed miRNAs between cumulus (CCs) and Mural (MGCs) Granulosa Cells. n=3 individual patients.
Fig. (3)
Fig. (3)
RT-qPCR validation of differentially expressed miRNAs between MGCs and CCs as identified by small RNA sequencing. Closed circles represent the three patients used for small RNA sequencing and open circles represent four additional patients. n= 7 individual patients in total. ** p <0.01, *** p < 0.001.
See this image and copyright information in PMC

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