scholarly journals Deletion of Gremlin-2 alters estrous cyclicity and disrupts female fertility in mice

2021 ◽  
Author(s):  
Robert T Rydze ◽  
Bethany Patton ◽  
Shawn M Briley ◽  
Hannia Salazar-Torralba ◽  
Gregory Gipson ◽  
...  
Keyword(s):  
Ovarian Follicle ◽  
Primary Ovarian Insufficiency ◽  
Protein Family ◽  
Bmp Signaling ◽  
Wild Type ◽  
Female Reproduction ◽  
Ovarian Insufficiency ◽  
Primordial Follicles ◽  
Ovarian Follicle Development ◽  
Wild Type Female

Abstract Members of the differential screening-selected gene aberrative in neuroblastoma (DAN) protein family are developmentally conserved extracellular binding proteins that antagonize bone morphogenetic protein (BMP) signaling. This protein family includes the Gremlin proteins, GREM1 and GREM2, which have key functions during embryogenesis and adult physiology. While BMPs play essential roles in ovarian follicle development, the role of the DAN family in female reproductive physiology is less understood. We generated mice null for Grem2 to determine its role in female reproduction in addition to screening patients with primary ovarian insufficiency for variants in GREM2. Grem2−/− mice are viable, but female Grem2−/− mice have diminished fecundity and irregular estrous cycles. This is accompanied by significantly reduced production of ovarian anti-Müllerian hormone (AMH) from small growing follicles, leading to a significant decrease in serum AMH. Surprisingly, as AMH is a well-established marker of the ovarian reserve, morphometric analysis of ovarian follicles showed maintenance of primordial follicles in Grem2−/− mice like wild type littermates. While Grem2 mRNA transcripts were not detected in the pituitary, Grem2 is expressed in hypothalami of wild type female mice, suggesting the potential for dysfunction in multiple tissues composing the hypothalamic–pituitary-ovarian axis that contribute to the subfertility phenotype. Additionally, screening 106 women with primary ovarian insufficiency identified one individual with a heterozygous variant in GREM2 that lies within the predicted BMP-GREM2 interface. In total, these data suggest Grem2 is necessary for female fecundity by playing a novel role in regulating the HPO axis and contributing to female reproductive disease.

scholarly journals Deletion of Gremlin-2 alters estrous cyclicity and disrupts female fertility in mice

2020 ◽  
Author(s):  
Robert T. Rydze ◽  
Bethany Patton ◽  
Hannia Salazar-Torralba ◽  
Shawn Briley ◽  
Gregory Gipson ◽  
...  
Keyword(s):  
Ovarian Function ◽  
Serum Levels ◽  
Primary Ovarian Insufficiency ◽  
Protein Family ◽  
Bmp Signaling ◽  
Female Fertility ◽  
Ovarian Insufficiency ◽  
Adult Mice ◽  
Morphogenetic Protein ◽  
Wild Type Female

AbstractMembers of the differential screening-selected gene aberrative in neuroblastoma (DAN) protein family are developmentally conserved extracellular binding proteins that antagonize bone morphogenetic protein (BMP) signaling. This protein family includes the Gremlin proteins, GREM1 and GREM2, which are known to have key functions during embryogenesis and adult physiology. While BMPs play essential roles in adult female reproductive physiology, the role of the DAN family in ovarian function is less understood. We generated mice null for Grem2 to study its role in female fertility in addition to screening patients with primary ovarian insufficiency for variants in GREM2. Grem2-/- mice are viable and female Grem2-/- mice have diminished fecundity and irregular estrous cycles. This is accompanied by reduced serum levels of anti-Müllerian hormone, a marker of the ovarian reserve, in adult mice. Alterations in ovarian expression of inhibin and activin subunit genes, which are required for regulation of the hypothalamic-pituitary-ovarian (HPO) axis, were identified. While Grem2 mRNA transcript was not detected in the pituitary, Grem2 was expressed in the hypothalami of wild type female mice. Additionally, screening 106 women with primary ovarian insufficiency identified one individual with a heterozygous variant in GREM2 that lies within the predicted BMP-GREM2 interface. In total, these data suggest that Grem2 is necessary for female fecundity by playing a novel role in regulating the HPO axis and possibly contributing to female reproductive disease.

scholarly journals Anti-Müllerian Hormone Attenuates the Effects of FSH on Follicle Development in the Mouse Ovary

Endocrinology ◽  
2001 ◽  
Vol 142 (11) ◽  
pp. 4891-4899 ◽  
Author(s):  
Alexandra L. L. Durlinger ◽  
Maria J. G. Gruijters ◽  
Piet Kramer ◽  
Bas Karels ◽  
T. Rajendra Kumar ◽  
...  
Keyword(s):  
Ovarian Follicle ◽  
Inhibitory Effect ◽  
Ovarian Follicles ◽  
Follicle Development ◽  
Wild Type ◽  
Follicle Growth ◽  
Primordial Follicles ◽  
Ovarian Follicle Development

Abstract Although ovarian follicle growth is under the influence of many growth factors and hormones of which FSH remains one of the most prominent regulators. Therefore, factors affecting the sensitivity of ovarian follicles to FSH are also important for follicle growth. The aim of the present study was to investigate whether anti-Müllerian hormone (AMH) has an inhibitory effect on follicle growth by decreasing the sensitivity of ovarian follicles to FSH. Furthermore, the combined action of AMH and FSH on ovarian follicle development was examined. Three different experiments were performed. Using an in vitro follicle culture system it was shown that FSH-stimulated preantral follicle growth is attenuated in the presence of AMH. This observation was confirmed by an in vivo experiment showing that in immature AMH-deficient females, more follicles start to grow under the influence of exogenous FSH than in their wild-type littermates. In a third experiment, examination of the follicle population of 4-month-old wild-type, FSHβ-, AMH-, and AMH-/FSHβ-deficient females revealed that loss of FSH expression has no impact on the number of primordial and preantral follicles, but the loss of inhibitory action of AMH on the recruitment of primordial follicles in AMH-deficient mice is increased in the absence of FSH. In conclusion, these studies show that AMH inhibits FSH-stimulated follicle growth in the mouse, suggesting that AMH is one of the factors determining the sensitivity of ovarian follicles for FSH and that AMH is a dominant regulator of early follicle growth.

scholarly journals Homozygous Inactivating Mutation inNANOS3in Two Sisters with Primary Ovarian Insufficiency

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Mariza G. Santos ◽  
Aline Z. Machado ◽  
Conceição N. Martins ◽  
Sorahia Domenice ◽  
Elaine M. F. Costa ◽  
...  
Keyword(s):  
Rna Binding ◽  
Mutational Analysis ◽  
Primordial Germ Cells ◽  
Embryonic Cell ◽  
Primary Ovarian Insufficiency ◽  
Primary Amenorrhea ◽  
Female Reproduction ◽  
Ovarian Insufficiency ◽  
Rna Interaction

Despite the increasing understanding of female reproduction, the molecular diagnosis of primary ovarian insufficiency (POI) is seldom obtained. The RNA-binding protein NANOS3 poses as an interesting candidate gene for POI since members of the Nanos family have an evolutionarily conserved function in germ cell development and maintenance by repressing apoptosis. We performed mutational analysis ofNANOS3in a cohort of 85 Brazilian women with familial or isolated POI, presenting with primary or secondary amenorrhea, and in ethnically-matched control women. A homozygous p.Glu120Lys mutation inNANOS3was identified in two sisters with primary amenorrhea. The substituted amino acid is located within the second C2HC motif in the conserved zinc finger domain of NANOS3 andin silicomolecular modelling suggests destabilization of protein-RNA interaction.In vitroanalyses of apoptosis through flow cytometry and confocal microscopy show that NANOS3 capacity to prevent apoptosis was impaired by this mutation. The identification of an inactivating missense mutation inNANOS3suggests a mechanism for POI involving increased primordial germ cells (PGCs) apoptosis during embryonic cell migration and highlights the importance of NANOS proteins in human ovarian biology.

scholarly journals Conditional Deletion of FOXL2 and SMAD4 in Gonadotropes of Adult Mice Causes Isolated FSH Deficiency

Endocrinology ◽  
2018 ◽  
Vol 159 (7) ◽  
pp. 2641-2655 ◽  
Author(s):  
Yining Li ◽  
Gauthier Schang ◽  
Ying Wang ◽  
Xiang Zhou ◽  
Adrien Levasseur ◽  
...  
Keyword(s):  
Ovarian Follicle ◽  
Conditional Knockout ◽  
Tamoxifen Treatment ◽  
Female Sterility ◽  
Follicle Development ◽  
Female Reproduction ◽  
Ovarian Follicle Development ◽  
Adult Mice ◽  
Conditional Deletion ◽  
Driver Line

Abstract The glycoprotein FSH, a product of pituitary gonadotrope cells, regulates ovarian follicle development in females and spermatogenesis in males. FSH is a heterodimer of the common α gonadotropin subunit and the hormone-specific FSHβ subunit (a product of the Fshb gene). Using a conditional knockout approach (Cre-lox), we previously demonstrated that Fshb expression in mice depends on the transcription factors forkhead box L2 (FOXL2) and SMAD4. Deletion of Foxl2 or Smad4 alone led to FSH deficiency, female subfertility, and oligozoospermia in males. Simultaneous deletion of the two genes yielded a greater suppression of FSH and female sterility. The Cre-driver used previously was first active during embryonic development. Therefore, it is unclear whether FOXL2 and SMAD4 play important roles in the development or adult function of gonadotropes, or both. To address this question, we developed a tamoxifen-inducible Cre-driver line, which enabled Foxl2 and Smad4 gene deletions in gonadotropes of adult mice. After tamoxifen treatment, females with previously demonstrated fertility exhibited profound reductions in FSH levels, arrested ovarian follicle development, and sterility. FSH levels were comparably reduced in males 1 or 2 months after treatment; however, spermatogenesis was unaffected. These data indicate that (1) FOXL2 and SMAD4 are necessary to maintain FSH synthesis in gonadotrope cells of adult mice, (2) FSH is essential for female reproduction but appears to be unnecessary for the maintenance of spermatogenesis in adult male mice, and (3) the inducible Cre-driver line developed here provides a powerful tool to interrogate gene function in gonadotrope cells of adult mice.

scholarly journals CPEB3 deficiency in mice affect ovarian follicle development and causes premature ovarian insufficiency

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Fang E. ◽  
He Zhang ◽  
Wanli Yin ◽  
Chongyang Wang ◽  
Yuanli Liu ◽  
...  
Keyword(s):  
Rna Binding ◽  
Binding Protein ◽  
Ovarian Follicle ◽  
Premature Ovarian Insufficiency ◽  
Follicle Development ◽  
Ovarian Insufficiency ◽  
Rna Sequences ◽  
Second Child ◽  
Ovarian Follicle Development ◽  
Element Binding Protein

AbstractPremature ovarian insufficiency (POI) is a heterogeneous and multifactorial disorder. In recent years, there has been an increasing interest in research on the pathogenesis and treatment of POI, owing to the implementation of the second-child policy in China. Cytoplasmic polyadenylation element-binding protein 3 (CPEB3) is an RNA-binding protein that can bind to specific RNA sequences. CPEB3 can bind to and affect the expression, cellular location, and stability of target RNAs. Cpeb3 is highly expressed in the ovary; however, its functions remain unknown. In this study, Cpeb3-mutant mice were used to characterize the physiological functions of CPEB3. Cpeb3-mutant female mice manifested signs of gradual loss of ovarian follicles, ovarian follicle development arrest, increased follicle atresia, and subfertility with a phenotype analogous to POI in women. Further analysis showed that granulosa cell proliferation was inhibited and apoptosis was markedly increased in Cpeb3-mutant ovaries. In addition, the expression of Gdf9, a potential target of CPEB3, was decreased in Cpeb3-mutant ovaries and oocytes. Altogether, these results reveal that CPEB3 is essential for ovarian follicle development and female fertility as it regulates the expression of Gdf9 in oocytes, disruption of which leads to impaired ovarian follicle development and POI.

scholarly journals Proteomic Analysis of Fetal Ovaries Reveals That Primordial Follicle Formation and Transition Are Differentially Regulated

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Mengmeng Xu ◽  
Long Che ◽  
Zhenguo Yang ◽  
Pan Zhang ◽  
Jiankai Shi ◽  
...  
Keyword(s):  
Stress Response ◽  
Molecular Mechanism ◽  
Expression Profiles ◽  
Ovarian Follicle ◽  
Primordial Follicle ◽  
Reproductive Organ ◽  
Primary Follicle ◽  
Altered Expression ◽  
Primordial Follicles ◽  
Ovarian Follicle Development

Primordial follicle formation represents a critical phase of the initiation of embryonic reproductive organ development, while the primordial follicle transition into primary follicle determines whether oestrus or ovulation will occur in female animals. To identify molecular mechanism of new proteins which are involved in ovarian development, we employed 2D-DIGE to compare the protein expression profiles of primordial follicles and primary follicles of fetal ovaries in pigs. Fetal ovaries were collected at distinct time-points of the gestation cycle (g55 and g90). The identified proteins at the g55 time-point are mainly involved in the development of anatomical structures [reticulocalbin-1 (RCN1), reticulocalbin-3 (RCN3)], cell differentiation (actin), and stress response [heterogeneous nuclear ribonucleoprotein K (HNRNPK)]. Meanwhile, at the g90 stage, the isolated proteins with altered expression levels were mainly associated with cell proliferation [major vault protein (MVP)] and stress response [heat shock-related 70 kDa protein 2 (HSPA2)]. In conclusion, our work revealed that primordial follicle formation is regulated by RCN1, RCN3, actin, and HNRNPK, while the primordial follicle transformation to primary follicle is regulated by MVP and HSPA2. Therefore, our results provide further information for the prospective understanding of the molecular mechanism(s) involved in the regulation of the ovarian follicle development.

scholarly journals Anti-Müllerian hormone overexpression restricts preantral ovarian follicle survival

2018 ◽  
Vol 237 (2) ◽  
pp. 153-163 ◽  
Author(s):  
Michael W Pankhurst ◽  
Rebecca L Kelley ◽  
Rachel L Sanders ◽  
Savana R Woodcock ◽  
Dorothy E Oorschot ◽  
...  
Keyword(s):  
Reproductive Output ◽  
Ovarian Follicle ◽  
Follicle Stimulating Hormone ◽  
Primordial Follicle ◽  
Slight Reduction ◽  
Wild Type ◽  
Primordial Follicles ◽  
Antral Follicles ◽  
Primordial Follicle Activation ◽  
Ovarian Folliculogenesis

Anti-Müllerian hormone (AMH) is an ovarian regulator that affects folliculogenesis. AMH inhibits the developmental activation of the dormant primordial follicles and the oocyte within. In more mature follicles, AMH reduces granulosa cell sensitivity to follicle-stimulating hormone (FSH). We examined the effects of AMH overexpression on the stages of ovarian folliculogenesis, and the development of embryos, with a transgenic mouse that overexpresses human AMH in central nervous system neurons under the control of the mouse Thy1.2 promoter (Thy1.2-AMH Tg mice). These mice are severely sub-fertile, despite relatively normal ovulation rates. The embryos of Thy1.2-AMHTg females exhibited delayed preimplantation development and extensive mid-gestation fetal resorption. Young Thy1.2-AMHTg mouse ovaries exhibited only a slight reduction in the rate of primordial follicle activation but large declines in the number of developing follicles surviving past the primary stage. It was expected that Thy1.2-AMHTg mice would retain more primordial follicles as they aged, but at 5 months, their number was significantly reduced relative to wild-type females. These data indicate that moderate elevations in AMH levels can severely restrict reproductive output and the number of developing follicles in the ovary. This evidence suggests that early antral follicles are a target for AMH signaling, which may regulate early follicle survival.

scholarly journals A homozygous FANCM mutation underlies a familial case of non-syndromic primary ovarian insufficiency

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Baptiste Fouquet ◽  
Patrycja Pawlikowska ◽  
Sandrine Caburet ◽  
Celine Guigon ◽  
Marika Mäkinen ◽  
...  
Keyword(s):  
Dna Damage Response ◽  
Meiotic Recombination ◽  
Chromosomal Abnormalities ◽  
Primary Ovarian Insufficiency ◽  
Homozygous Mutation ◽  
Wild Type ◽  
Ovarian Insufficiency ◽  
Response Gene ◽  
Damage Response ◽  
Fancd2 Monoubiquitination

Primary Ovarian Insufficiency (POI) affects ~1% of women under forty. Exome sequencing of two Finnish sisters with non-syndromic POI revealed a homozygous mutation in FANCM, leading to a truncated protein (p.Gln1701*). FANCM is a DNA-damage response gene whose heterozygous mutations predispose to breast cancer. Compared to the mother's cells, the patients’ lymphocytes displayed higher levels of basal and mitomycin C (MMC)-induced chromosomal abnormalities. Their lymphoblasts were hypersensitive to MMC and MMC-induced monoubiquitination of FANCD2 was impaired. Genetic complementation of patient's cells with wild-type FANCM improved their resistance to MMC re-establishing FANCD2 monoubiquitination. FANCM was more strongly expressed in human fetal germ cells than in somatic cells. FANCM protein was preferentially expressed along the chromosomes in pachytene cells, which undergo meiotic recombination. This mutation may provoke meiotic defects leading to a depleted follicular stock, as in Fancm-/- mice. Our findings document the first Mendelian phenotype due to a biallelic FANCM mutation.

scholarly journals Obesity alters the ovarian proteomic response to zearalenone exposure

2021 ◽  
Author(s):  
M Estefanía González Alvarez ◽  
Bailey C McGuire ◽  
Aileen F Keating
Keyword(s):  
Obese Mice ◽  
Wild Type ◽  
Female Reproduction ◽  
Primordial Follicles ◽  
Ovary Weight ◽  
Obese Female ◽  
Lethal Yellow ◽  
Estrous Cyclicity ◽  
17Β Estradiol ◽  
A Minor

Abstract Zearalenone (ZEN), a non-steroidal estrogenic mycotoxin, is detrimental to female reproduction. Altered chemical biotransformation, depleted primordial follicles and a blunted genotoxicant response have been discovered in obese female ovaries, thus, this study investigated the hypothesis that obesity would enhance ovarian sensitivity to ZEN exposure. Seven week old female wild type non-agouti KK.Cg-a/a mice (lean) and agouti lethal yellow KK.Cg-Ay/J mice (obese) received food and water ad libitum, and either saline or ZEN (40 μg/Kg) per os for 15 days. Body and organ weights, and estrous cyclicity were recorded, and ovaries collected post-euthanasia for protein analysis. Body and liver weights were increased (P < 0.05) in the obese mice, but obesity did not affect (P > 0.05) heart, kidney, spleen, uterus, or ovary weight and there was no impact (P > 0.05) of ZEN exposure on body or organ weight in lean or obese mice. Obese mice had shorter proestrus (P < 0.05) and a tendency (P = 0.055) for longer metestrus/diestrus. ZEN exposure in obese mice increased estrus but shortened metestrus/diestrus length. Neither obesity or ZEN exposure impacted (P > 0.05) circulating 17β-estradiol or progesterone, or ovarian abundance of EPHX1, GSTP1, CYP2E1, ATM, BRCA1, DNMT1, HDAC1, H4K16ac, or H3K9me3. Lean mice exposed to ZEN had a minor increase in γH2AX abundance (P < 0.05). In lean and obese mice, LC–MS/MS identified alterations to proteins involved in chemical metabolism, DNA repair and reproduction. These data identify ZEN-induced adverse ovarian modes of action and suggest that obesity is additive to ZEN-induced ovotoxicity.

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