Abstract:
STUDY QUESTION:Can RNA sequencing of human cumulus cells (CC) reveal molecular pathways involved in the physiology of reproductive aging? STUDY FINDING:Senescent but not young CC activate gene pathways associated with hypoxia and oxidative stress. WHAT IS KNOWN ALREADY:Shifts in socioeconomic norms are resulting in larger numbers of women postponing childbearing. The reproductive potential is sharply decreased with aging, and the reasons are poorly understood. Since CCs play an integral role in oocyte maturation and direct access to human oocytes is limited, we used whole transcriptome analysis of these somatic cells to gain insights into the molecular mechanisms playing a role in follicular senescence. STUDY DESIGN, SAMPLES/MATERIALS, METHODS:Twenty CC samples (from a total of 15 patients) were obtained from oocytes of either male factor or egg donor patients. RNA sequencing and bioinformatic tools were used to identify differentially expressed genes between CCs from seven aged and eight young patients (<35 (years old) y.o. vs >40 y.o.). Quantitative-PCR and immunoflourescent staining were used for validation. MAIN RESULTS AND THE ROLE OF CHANCE:RNA sequencing identified 11 572 genes expressed in CC of both age cohorts, 45 of which were differentially expressed. In CC collected from patients >40 y.o., genes involved in the hypoxia stress response (NOS2, RORA and NR4A3), vasculature development (NR2F2, PTHLH), glycolysis (RALGAPA2 and TBC1D4) and cAMP turnover (PDE4D) were significantly overexpressed when compared with CC of patients younger than 35 y.o. LIMITATIONS, REASONS FOR CAUTION:This study focused almost exclusively on assessing the genetic differences in CC transcriptome between young and older women. These genetic findings were not fully correlated with embryonic development and clinical outcome. WIDER IMPLICATIONS OF THE FINDINGS:Our data provide a new hypothesis-follicular hypoxia-as the main mechanism leading to ovarian follicular senescence and suggest a link between cumulus cell aging and oocyte quality decay. If specific molecular findings of hypoxia would be confirmed also in oocytes, genetic platforms could screen CC for hypoxic damage and identify healthier oocytes. Protocols of ovarian stimulation in older patients could also be adjusted to diminish oocyte exposure time to hypoxic follicles. LARGE SCALE DATA:GEO accession number: GSE81579 STUDY FUNDING AND COMPETING INTERESTS: Funded in part by EMD Serono Grant for Fertility Innovation (GFI).
journal_name
Mol Hum Reprodjournal_title
Molecular human reproductionauthors
Molinari E,Bar H,Pyle AM,Patrizio Pdoi
10.1093/molehr/gaw038subject
Has Abstractpub_date
2016-08-01 00:00:00pages
866-76issue
8eissn
1360-9947issn
1460-2407pii
gaw038journal_volume
22pub_type
杂志文章abstract::Microarray studies generating lists of genes with altered expression in placentas from pregnancies complicated with pre-eclampsia (PE) have so far been published in several different studies. Working under the assumption that altered gene expression in PE may be the result of altered expression of regulatory transcrip...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gal001
更新日期:2006-01-01 00:00:00
abstract::Human sperm contain similar amounts of protamine-1 (P1) and protamine-2 (P2). Although an aberrant protamine ratio have been observed in subfertile men, functional evidence is provided by protamine knockout mice exhibiting male infertility. As sperm DNA integrity is known to be linked with DNA fragmentation and apopto...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gar011
更新日期:2011-07-01 00:00:00
abstract::Embryo implantation and subsequent decidualization, trophoblast invasion and formation of a functional placenta are crucial for establishment and maintenance of pregnancy. Interleukin-11 signalling has been shown to be obligatory for adequate decidualization and trophoblast invasion in mice. Defects in IL-11 signallin...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gah107
更新日期:2004-11-01 00:00:00
abstract::Folate deficiency has been linked to a wide range of pregnancy disorders. Most research about folate-deficiency has focused on the embryo itself, little attention has been paid to possible effects on the placenta. According to our results, the morphology of the placenta, endocrine function, and the expression of genes...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gaz022
更新日期:2019-06-06 00:00:00
abstract::The Y chromosome carries several spermatogenesis genes distributed in three regions: AZFa, AZFb and AZFc. Microdeletions in these regions have been seen in 10% of sterile males with azoospermia or oligozoospermia, the most frequent of them being characterized by a complete deletion of AZFc region. A partial AZFc delet...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gal029
更新日期:2006-04-01 00:00:00
abstract::Glutathione S-transferase M1 (GSTM1), one member of the GST family, is responsible for metabolism of xenobiotics and carcinogens. Myeloperoxidase (MPO) plays an important role in the oxidation and activation of carcinogens and nitric oxide. Allelic variants of GSTM1 and MPO gene polymorphisms might impair detoxificati...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gah095
更新日期:2004-10-01 00:00:00
abstract::Sperm are motile cells. Thus, a significant component of the spermatogenic cycle is devoted to the formation of flagellum, a process that must be coordinated to insure proper construction. To document the temporal pattern of flagellar gene expression, we employed real-time PCR to assess changes in accumulation of a co...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gah163
更新日期:2005-04-01 00:00:00
abstract::The 2019 novel coronavirus (2019-nCoV) appeared in December 2019 and then spread throughout the world rapidly. The virus invades the target cell by binding to angiotensin-converting enzyme (ACE) 2 and modulates the expression of ACE2 in host cells. ACE2, a pivotal component of the renin-angiotensin system, exerts its ...
journal_title:Molecular human reproduction
pub_type: 杂志文章,评审
doi:10.1093/molehr/gaaa030
更新日期:2020-06-01 00:00:00
abstract::Natural killer (NK) cells are the predominant leukocyte in first trimester decidua and play a role in vascular remodelling through interferon gamma (IFNgamma) secretion. Membrane expression of the killer immunoglobulin-like receptor (KIR) KIR2DL4 on peripheral blood NK (pNK) cells is controlled by the 9A/10A transmemb...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gap039
更新日期:2009-08-01 00:00:00
abstract::We have previously reported that endometrial expression of matrix metalloproteinase (MMP)-26 mRNA comes to a maximum in the early secretory phase. Since tissue inhibitor of metalloproteinase (TIMP)-4 is a potent inhibitor of MMP-26, the objective of this study was to identify the pattern of TIMP-4 mRNA expression in t...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gah092
更新日期:2004-09-01 00:00:00
abstract::Oocyte vitrification has been introduced into clinical settings without extensive pre-clinical safety testing. In this study, we analysed major safety aspects of human oocyte vitrification in a high security closed system: (i) chromosomal meiotic segregation, (ii) embryonic developmental kinetics and (iii) DNA (hydrox...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gav013
更新日期:2015-06-01 00:00:00
abstract::The endocrine disrupting chemical o, p'-dichlorodiphenyltrichloroethane (DDT) can affect reproductive organs, tissues and cells in several species. Treatment of human endometrial endothelial cells (HEECs) with 50 microM o,p'-DDT decreased their proliferation compared with the control. Microarray analyses revealed that...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gam091
更新日期:2008-02-01 00:00:00
abstract::Several studies have demonstrated an association between polycystic ovary syndrome (PCOS) and the dinucleotide repeat microsatellite marker D19S884, which is located in intron 55 of the fibrillin-3 (FBN3) gene. Fibrillins, including FBN1 and 2, interact with latent transforming growth factor (TGF)-beta-binding protein...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gap072
更新日期:2009-12-01 00:00:00
abstract::In oocytes, cytoplasmic 3' polyadenylation regulates translational activation of dormant mRNA during meiotic maturation. Thus exogenous proteins are hardly expressed after injection of conventional RNA. To circumvent this, we synthesized a long polyadenylated (approximately 250 A) tail to encode RNA with an enhanced y...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/7.11.1039
更新日期:2001-11-01 00:00:00
abstract::We purified by fractionation on 10-40% glycerol gradients, 26S proteasomes from normal human spermatozoa. These proteasomes, which participate in the ATP-dependent degradation of ubiquitinated proteins, share a similar sedimentation coefficient to those purified from other human tissues. Fluorogenic peptide assays rev...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/3.12.1053
更新日期:1997-12-01 00:00:00
abstract::Nitric oxide (NO) is synthesized from L-arginine by a family of enzymes known as the nitric oxide synthases (NOS). We have recently shown a NOS similar to constitutive brain NOS (bNOS) and endothelial NOS (ecNOS) to be present in spermatozoa. The aim of this study is to investigate NO production by human spermatozoa a...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/3.9.755
更新日期:1997-09-01 00:00:00
abstract::Retrograde flow of menstrual blood cells during menstruation is considered as the dominant theory for the development of endometriosis. Moreover, current evidence suggests that endometrial-derived stem cells are key players in the pathogenesis of endometriosis. In particular, endometrial stromal stem cells have been s...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gau044
更新日期:2014-09-01 00:00:00
abstract::Mouse cauda epididymis were in-vivo transfected using the lipid FuGENE 6 as gene vector. Two gene constructions were employed: the p-GeneGRIP which codifies for the Green Fluorescent Protein (GFP) and the pSEAP-control that expresses an alkaline phosphatase as a secretion. Transfection was detected by fluorescence and...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gap026
更新日期:2009-06-01 00:00:00
abstract::Successful pregnancy in humans depends on deep invasion of the maternal decidua by extravillous trophoblast cells (EVTs), a process regulated by autocrine and paracrine signals in the decidual-trophoblast microenvironment. Here we examined whether trophoblast invasion is affected by decidual differentiation of endomet...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gar015
更新日期:2011-07-01 00:00:00
abstract::The androgen receptor (AR) gene is located on the X chromosome and contains a polymorphic CAG tract. CAG repeat expansions in the AR have been associated with male infertility and the neuromuscular disease, spinal bulbar muscular atrophy (SBMA). Based on Mendelian inheritance patterns, moderate CAG expansions in infer...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/6.9.861
更新日期:2000-09-01 00:00:00
abstract::Analysis of small numbers of chromosomes using interphase fluorescent in-situ hybridization (FISH) probes has revealed that 50% of human preimplantation embryos contain abnormal cells. Detection of high levels of mosaicism with so few probes has led some researchers to extrapolate that a full analysis of all 23 pairs ...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/6.11.1055
更新日期:2000-11-01 00:00:00
abstract::This study was undertaken to localize ryanodine-sensitive Ca2+ stores in human oocytes and to evaluate their role in the Ca2+ oscillations responsible for oocyte activation at fertilization. The addition of ryanodine provoked a Ca2+ discharge from stores localized throughout the ooplasm with the exception of the corti...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/2.4.265
更新日期:1996-04-01 00:00:00
abstract:STUDY QUESTION:Do human adult Leydig cells (ALCs) within hyperplastic micronodules display characteristics of foetal LCs (FLCs)? SUMMARY ANSWER:The gene expression profiles of FLCs and all ALC subgroups were clearly different, but there were no significant differences in expressed genes between the normally clustered ...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gax012
更新日期:2017-05-01 00:00:00
abstract::Glycoforms of recombinant human follicle stimulating hormone (rhFSH) (Org 32489, Puregon) were characterized using concanavalin A lectin affinity chromatography to reveal information about the internal carbohydrate complexity (extent of carbohydrate side-chain branching) of the preparations. The rhFSH glycoforms were ...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/2.10.807
更新日期:1996-10-01 00:00:00
abstract::SRY directs testicular development. It has been suggested that the only high-mobility group (HMG) box of the SRY is important for the function of this protein; however, other studies have suggested that the N- and C-terminal regions are also involved in this process. Herein, we analysed and compared in vitro the DNA-b...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gan021
更新日期:2008-06-01 00:00:00
abstract:STUDY HYPOTHESIS:In women with preterm premature rupture of the membranes (PPROM), increased oxidative stress may accelerate premature cellular senescence, senescence-associated inflammation and proteolysis, which may predispose them to rupture. STUDY FINDING:We demonstrate mechanistic differences between preterm birt...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gav074
更新日期:2016-02-01 00:00:00
abstract::In mammals, egg activation is initiated by multiple cytosolic Ca(2+) transients (Ca(2+) oscillations) that are triggered following delivery of a putative sperm factor from the fertilizing sperm. The identity of this 'sperm factor' thus holds much significance, not only as a vital component in creating a new life, but ...
journal_title:Molecular human reproduction
pub_type: 杂志文章,评审
doi:10.1093/molehr/gav009
更新日期:2015-05-01 00:00:00
abstract::We have developed a competitive reverse transcription-polymerase chain reaction (RT-PCR) sensitive enough to detect and quantify as little as 2-fold differences in gene expression in individual oocytes and embryos throughout human preimplantation development. This RT-PCR assay can be tailored for the examination of an...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/7.2.147
更新日期:2001-02-01 00:00:00
abstract::Human endometrial stromal cells (ESC) can produce a variety of chemokines, especially after inflammatory stimulation. Interferon-gamma-inducible protein-10 (IP-10) is a potent chemoattractant for lymphocytes, and belongs to the family of non-ELR CXC chemokines. The expression of IP-10 in ESC after stimulation with int...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/8.2.176
更新日期:2002-02-01 00:00:00
abstract::An understanding of the relationship between nuclear morphology and DNA function is important in cytology and preimplantation diagnosis. In this study, direct polymerase chain reaction (PCR) amplification was used to diagnose the common delta F508 mutation of cystic fibrosis in 62 biopsied human embryo cells. The nucl...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/2.1.63
更新日期:1996-01-01 00:00:00