Dormancy and activation of human oocytes from primordial and primary follicles: molecular clues to oocyte regulation

Ernst, Emil Hagen; Grøndahl, Marie Louise; Grund, Silvia K.; Hardy, Kate; Heuck, Anders; Sunde, Lone; Franks, Stephen; Andersen, Claus Yding; Villesen, Palle; Lykke‐Hartmann, Karin

doi:10.1093/humrep/dex238

Cited by 85 publications

(110 citation statements)

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“…In mouse ovary, Rac1 is involved in primordial follicle formation by inducing nuclear transport of SMAD3 29 . As mentioned above, PI3K-AKT signaling pathway have crucial roles to balance follicle growth suppression, activation and progression also in humans 30 and BMPs have important role in follicle development including primordial germ cell development, oocyte-somatic cell interactions and modulating cumulus-oocyte complex (COC) formation and expansion 31 .…”

Section: Discussionmentioning

confidence: 99%

Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells

Rooda

Hensen

Kaselt

et al. 2020

Sci Rep

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MicroRNAs (miRNAs) are known post-transcriptional regulators of various biological processesincluding ovarian follicle development. We have previously identified miRNAs from human preovulatory ovarian granulosa cells that are expressed from the intronic regions of two key genes in normal follicular development: FSH receptor (FSHR) and CYP19A1, the latter encoding the aromatase enzyme. The present study aims to identify the target genes regulated by these miRNAs: hsa-miR-548ba and hsa-miR-7973, respectively. The miRNAs of interest were transfected into KGN cell line and the gene expression changes were analyzed by Affymetrix microarray. Potential miRNA-regulated genes were further filtered by bioinformatic target prediction algorithms and validated for direct miRNA:mRNA binding by luciferase reporter assay. LIFR, PTEN, NEO1 and SP110 were confirmed as targets for hsa-miR-548ba. Hsa-miR-7973 target genes ADAM19, PXDN and FMNL3 also passed all verification steps. Additionally, the expression pattern of the miRNAs was studied in human primary cumulus granulosa cell culture in relation to the expression of their host genes and fSH stimulation. Based on our findings we propose the involvement of hsa-miR-548ba in the regulation of follicle growth and activation via LIFR and PTEN. Hsa-miR-7973 may be implicated in the modulation of extracellular matrix and cell-cell interactions by regulating the expression of its identified targets.Granulosa cell functions are essential in follicular development, maturation, and atresia. Granulosa cells support oocyte growth via continuous bidirectional communication to ensure oocyte quality and developmental competence. Due to the close communication cumulus (CGC) and mural granulosa cells (MGC) may reflect the characteristics of the oocyte and an understanding of the patterns of expression and functions of miRNAs in those cells may lead to a better understanding of follicle maturation and its dysfunction 1,2 .MicroRNAs (miRNAs) are short non-coding RNA molecules approximately 22 nucleotides in length 3 . MicroRNAs bind to their target mRNA 3′ untranslated region (3′UTR) at sites complementary to the miRNA 5′ seed region. MicroRNA biding to the target gene regulates gene expression by destabilizing the mRNA and/or inhibiting its translation 4 . In a few cases, an increase in gene expression has also been observed 5 .MicroRNAs have key role in the post-transcriptional regulation of various important biological processes, including cell proliferation, differentiation, apoptosis, and hormone biosynthesis and secretion 6 . Functional ovary requires precise coordination of follicle recruitment, selection and ovulation processes. Development of ovarian follicles is a complex process including oocyte maturation, granulosa cell proliferation and differentiation. MicroRNAs expressed in the ovary have regulative roles in ovarian follicle development and the expression of miRNAs in the ovary varies within the specific cell type and function 7 . The overall role of miRNAs in ovarian functions has ...

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Section: Discussionmentioning

confidence: 99%

Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells

Rooda

Hensen

Kaselt

et al. 2020

Sci Rep

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show abstract

“…In mouse ovary, Rac1 is involved in primordial follicle formation by inducing nuclear transport of SMAD3 [53]. As mentioned above, PI3K-AKT signaling pathway have crucial roles to balance follicle growth suppression, activation and progression also in humans [54] and BMPs have important role in follicle development including primordial germ cell development, oocyte-somatic cell interactions and modulating of COC formation and expansion [55].…”

Section: Discussionmentioning

confidence: 99%

MicroRNAs expressed from FSHR and aromatase genes target important ovarian functions

Rooda

Kaselt

Salumets

et al. 2019

Preprint

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sentence: Confirmed targets of miRNAs hsa-miR-548ba and hsa-miR-7973 are 4 involved in follicle recruitment, apoptosis, intercellular interactions and extracellular matrix 5 remodeling pathways in KGN cells. 6 7 Ilmatar Abstract 29 MicroRNAs (miRNAs) have known roles in the post-transcriptional regulation of various 30 biological processes including ovarian follicle development. We have previously identified 31 miRNAs from human pre-ovulatory granulosa cells that are expressed from the intronic regions 32 of two key genes in normal follicular development: FSH receptor (FSHR) and CYP19A1, the 33 latter encoding the aromatase enzyme. In the present study, we aim to identify the targets 34 regulated by those two miRNAs: hsa-miR-548ba and hsa-miR-7973, respectively. The 35 miRNAs of interest were endogenously expressed in KGN cell-line, gene expression changes 36 were analyzed by Affymetrix microarray and confirmed by RT-qPCR. Potential miRNA-37 regulated sequences were further filtered from the obtained results by bioinformatic target 38 prediction algorithms and validated for direct miRNA:mRNA binding by luciferase reporter 39 assay. Our results verified Leukemia inhibitory factor receptor (LIFR), Phosphatase and tensin 40 homolog (PTEN), Neogenin 1 (NEO1) and SP110 nuclear body protein (SP110) as target genes 41 for hsa-miR-548ba. Hsa-miR-7973 target genes ADAM metallopeptidase domain 19 42 (ADAM19), Peroxidasin (PXDN) and Formin like 3 (FMNL3) also passed all verification 43 steps. In conclusion we propose that hsa-miR-548ba may be involved in the regulation of 44 follicle growth and activation via LIFR and PTEN. Hsa-miR-7973 may be implicated in the 45 modulation of extracellular matrix and cell-cell interactions. Taken together, our results 46 suggest that those two miRNAs of interest have important regulatory roles in granulosa cells 47 and in follicle development in general. 48 49

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“…Patients consented to the research conducted. Oocytes and follicles (oocytes with surrounding granulosa cells) from the primordial and primary stages respectively were isolated based on morphological appearance via laser capture microdissection (Veritas™ Microdissection Instrument Model 704, ArcturusXT™, Molecular Devices, Applied Biosystems, Life Technologies, Foster City, CA, USA) [41,43] from cryopreserved biopsies. Until use, the cortical piece was stored in liquid nitrogen (− 196°C), as previously described [44,45].…”

Section: Human Cortical Ovarian Tissue and Follicle Isolationmentioning

confidence: 99%

“…Ovarian cortical fragments with a size of 2 × 2 × 1 mm were thawed and fixed by direct immersion into 4% paraformaldehyde (PFA) at 4°C for 4 h followed by dehydration and embedding in paraffin, and LCM was performed using the Veritas™ Microdissection instrument Model 704 (ArcturusXT™, Molecular Devices, Applied Biosystems®, Life Technologies, Foster City, CA, USA), as described [46]. Isolates of oocytes and oocytes with surrounding granulosa cells (follicles) from the primordial and primary stage, respectively (four main groups in total), from each of the three patients, with a total number of 12 samples, were isolated using laser capture microdissection [41].…”

Section: Laser Capture Microdissectionmentioning

confidence: 99%

“…In silico extraction of granulosa cell transcriptomes was performed by applying three filters on global transcriptome data from patient triplicates of oocytes [46] and oocytes with surrounding granulosa cells (follicle) for both the primordial and primary stage [41]. First, consistency in mean gene expression level (FPKM) for all detected transcripts was quantified by performing a t test on patient triplicate samples of same type ((a) primordial oocyte, patient 1 + 2 + 3; (b) primordial follicle, patient 1 + 2 + 3; (c) primary oocyte, patient 1 + 2 + 3; and (d) primary follicle, patient 1 + 2 + 3).…”

Section: Library Preparation Sequencing and Bio-informatically Managmentioning

confidence: 99%

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Transcripts encoding free radical scavengers in human granulosa cells from primordial and primary ovarian follicles

Ernst

Lykke‐Hartmann

2018

J Assist Reprod Genet

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The results indicate extensive energy production and free radical scavenging in the granulosa cells of primordial follicles with potential implications for ovarian ageing, cigarette smoking, premature ovarian failure and polycystic ovarian syndrome. Furthermore, DJ-1 may be involved in androgen responsiveness and the regulation of follicle growth via PI3K/PTEN/AKT signalling pathway regulation in the granulosa cells of primary follicles. The involvement of mitochondrial free radical production in the age-related decline of competent oocytes is becoming apparent.

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Dormancy and activation of human oocytes from primordial and primary follicles: molecular clues to oocyte regulation

Cited by 85 publications

References 48 publications

Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells

Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells

MicroRNAs expressed from FSHR and aromatase genes target important ovarian functions

Transcripts encoding free radical scavengers in human granulosa cells from primordial and primary ovarian follicles

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