Granulosa cell and oocyte mitochondrial abnormalities in a mouse model of fragile X primary ovarian insufficiency

AbstractPrimary ovarian insufficiency (POI) is defined as the loss of ovarian function before 40 years of age. It clinically manifests as amenorrhea, infertility, and signs of estrogen insufficiency. POI is frequently induced by chemotherapy. Gonadotoxic chemotherapy reagents damage granulosa cells, which are essential for follicular function and development. Our recently published studies demonstrated that intraovarian transplantation of human mesenchymal stem cells (hMSCs) can restore fertility in a chemotherapy-induced POI mouse model. However, the regenerative mechanism underlying the hMSC effect in POI mice is not fully understood. Here, we report that the hMSC secretome increased the proliferation of human granulosa cells (HGrC1). We showed by FACS analysis that treatment of HGrC1 cells with hMSC-conditioned media (hMSC CM) stimulates cellular proliferation. We also demonstrated that the expression of steroidogenic enzymes involved in the production of estrogen, CYP19A1 and StAR, are significantly elevated in hMSC CM-treated HGrC1 cells. Our data suggest that hMSC CM stimulates granulosa cell proliferation and function, which may explain the therapeutic effect of hMSCs in our chemotherapy-induced POI animal model. Our findings indicate that the hMSC secretome may be a novel treatment approach for restoring granulosa cell and ovarian function in patients with POI.

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Establishment and Mechanism Study of a Primary Ovarian Insufficiency Mouse Model using Lipopolysaccharide

10.21203/rs.3.rs-545549/v1 ◽

2021 ◽

Author(s):

Si-Ji Lv ◽

Shu-Hui Hou ◽

Lei Gan ◽

Jing Sun

Keyword(s):

Mouse Model ◽

Granulosa Cell ◽

Cell Apoptosis ◽

Low Dose ◽

Ovarian Function ◽

Histopathological Examination ◽

Primary Ovarian Insufficiency ◽

High Dose ◽

Ovarian Insufficiency ◽

Primordial Follicles

Abstract Background: This study aimed to establish a lipopolysaccharide (LPS)-induced primary ovarian insufficiency (POI) mouse model and to investigate the underlying mechanism.Methods: C57BL/6N female mice were intraperitoneally injected with low-dose LPS (0.5 mg/kg) once daily for 14 days, high-dose LPS (2.5 mg/kg) twice weekly for 2 weeks, and cyclophosphamide (CTX; 150 mg/kg) once weekly for 2 weeks. Ovarian function was assessed by measuring the length of the estrous cycle, the number of primordial follicles, and the levels of serum pituitary/ovarian hormones. Expression and production of interleukin 1β (IL-1β) were determined to evaluate ovarian inflammation. Histopathological examination was performed to examine ovarian fibrosis. TUNEL assay was carried out to evaluate granulosa cell apoptosis. Western blotting was performed to measure the levels of inflammation-, fibrosis-, and apoptosis-related proteins in mouse ovaries.Results: Like CTX, both low- and high-dose LPS administration significantly impaired ovarian functions in mice, as evidenced by extended lengths of estrous cycles, reduced counts of primordial follicles, and alterations in the levels of serum hormones. Also, LPS administration promoted granulosa cell apoptosis and ovarian fibrosis in mice. However, LPS but not CTX significantly promoted IL-1β expression and production in mice. Moreover, LPS treatment but not CTX significantly enhanced TLR, p-p65, p65, and MyD88 protein expression in mouse ovaries, suggesting that LPS differs from CTX in triggering ovarian inflammation. In general, continuous low-dose LPS stimulation was less potent than high-dose LPS stimulation in the above-mentioned effects.Conclusions: LPS induces ovarian inflammation, fibrosis, and granulosa cell apoptosis and can be used to establish a POI model in mice.

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Ovarian Abnormalities in a Mouse Model of Fragile X Primary Ovarian Insufficiency

Journal of Histochemistry & Cytochemistry ◽

10.1369/0022155412441002 ◽

2012 ◽

Vol 60 (6) ◽

pp. 439-456 ◽

Cited By ~ 52

Author(s):

Gloria E. Hoffman ◽

Wei Wei Le ◽

Ali Entezam ◽

Noriyuki Otsuka ◽

Zhi-Bin Tong ◽

...

Keyword(s):

Mouse Model ◽

Fragile X ◽

Primary Ovarian Insufficiency ◽

Ovarian Insufficiency

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A mouse model for fragile X- associated primary ovarian insufficiency

Fertility and Sterility ◽

10.1016/j.fertnstert.2013.07.1841 ◽

2013 ◽

Vol 100 (3) ◽

pp. S52-S53

Author(s):

E. Mok-Lin ◽

M. Ascano ◽

Z. Rosenwaks ◽

T. Tuschl ◽

Z. Williams

Keyword(s):

Mouse Model ◽

Fragile X ◽

Primary Ovarian Insufficiency ◽

Ovarian Insufficiency

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Establishment and Mechanism Study of a Primary Ovarian Insufficiency Mouse Model Using Lipopolysaccharide

Analytical Cellular Pathology ◽

10.1155/2021/1781532 ◽

2021 ◽

Vol 2021 ◽

pp. 1-9

Author(s):

Si-Ji Lv ◽

Shu-Hui Hou ◽

Lei Gan ◽

Jing Sun

Keyword(s):

Mouse Model ◽

Granulosa Cell ◽

Cell Apoptosis ◽

Low Dose ◽

Histopathological Examination ◽

Primary Ovarian Insufficiency ◽

High Dose ◽

Ovarian Insufficiency ◽

Serum Hormones ◽

Primordial Follicles

This study is aimed at establishing a lipopolysaccharide- (LPS-) induced primary ovarian insufficiency (POI) mouse model and investigating the underlying mechanism. C57BL/6N female mice were intraperitoneally injected with low-dose LPS (0.5 mg/kg) once daily for 14 days, high-dose LPS (2.5 mg/kg) twice weekly for 2 weeks, or cyclophosphamide (CTX; 150 mg/kg) once weekly for 2 weeks. Ovarian function was assessed by measuring the length of estrous cycle, the number of primordial follicles, and the levels of serum hormones. Expression and production of interleukin 1β (IL-1β) were determined to evaluate ovarian inflammation. Histopathological examination was performed to examine ovarian fibrosis. TUNEL assay was carried out to evaluate granulosa cell apoptosis. Western blotting was performed to measure the levels of inflammation-, fibrosis-, and apoptosis-related proteins in the mouse ovaries. Like CTX, both low- and high-dose LPS significantly impaired ovarian functions in mice, as evidenced by extended lengths of estrous cycles, reduced counts of primordial follicles, and alterations in the levels of serum hormones. Also, LPS promoted granulosa cell apoptosis and ovarian fibrosis in mice. However, LPS but not CTX promoted IL-1β expression and production in mice. Moreover, LPS but not CTX enhanced TLR, p-p65, p65, and MyD88 expression in mouse ovaries, suggesting that LPS differs from CTX in triggering ovarian inflammation. In general, continuous low-dose LPS stimulation was less potent than high-dose LPS to affect the ovarian functions. In conclusion, LPS may induce ovarian inflammation, fibrosis, and granulosa cell apoptosis and can be used to establish a POI model in mice.

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Primary ovarian insufficiency in an adolescent population: clinical phenotype and diagnostic approach

Journal of Pediatric Endocrinology and Metabolism ◽

10.1515/jpem-2019-0083 ◽

2019 ◽

Vol 32 (9) ◽

pp. 979-985

Author(s):

Alfonso Hoyos-Martinez ◽

Luis R. Hoyos ◽

Metee Comkornruecha ◽

Alejandro Diaz

Keyword(s):

Clinical Phenotype ◽

Fragile X ◽

Clinical Information ◽

Rare Condition ◽

Primary Ovarian Insufficiency ◽

Diagnostic Approach ◽

Ovarian Insufficiency ◽

Adolescent Population ◽

Autoimmune Conditions ◽

Reproductive Counseling

Abstract Background Primary ovarian insufficiency (POI) can be seen in adolescents secondary to genetic or autoimmune conditions, or gonadotoxic therapies. Often times, its underlying cause is not identified. It is a rare condition in pediatrics, but a thorough evaluation is required for a timely diagnosis and optimizing outcomes. Objectives We aim to describe the clinical phenotype of idiopathic POI in an adolescent population seen in a referral center, and evaluate its diagnostic approach. Methods All patients evaluated between 2012 and 2018 were identified using the diagnostic codes for POI. Medical records were manually reviewed and clinical information was extracted. Cases were excluded from the final sample if they were found to have incomplete diagnostic information, Turner syndrome, eating disorders, gonadal surgeries and/or a history of oncological conditions or treatments. Results Forty-eight patients with POI were identified, and only seven met the established criteria. Anti-ovarian and anti-thyroid antibodies were evaluated in 100% and 86%, respectively, while only 29% were tested for anti-adrenal autoimmunity. The karyotype was obtained consistently, while the fragile X mental retardation 1 (FMR1) gene expansion was only assessed in approximately a third of the patients. Finally, only 29% of patients received reproductive counseling or referral to a fertility specialist. Conclusions Diagnostic evaluation for POI appears to be challenging to pediatric providers. Anti-ovarian antibodies are frequently obtained despite the lack of their clinical significance in POI, while anti-adrenal antibodies, which are the preferred diagnostic test, are not commonly obtained. Reproductive orientation or referral is seldom provided to the adolescent population.

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Assessment of FMR1 triplet repeats in patients affected with mental retardation, fragile X syndrome and primary ovarian insufficiency

Journal of Genetics ◽

10.1007/s12041-019-1171-5 ◽

2020 ◽

Vol 99 (1) ◽

Author(s):

Zeinab Salimy ◽

Mohammad Taghi Akbari ◽

Faravareh Khordadpoor Deilamani

Keyword(s):

Mental Retardation ◽

Fragile X Syndrome ◽

Fragile X ◽

Primary Ovarian Insufficiency ◽

Triplet Repeats ◽

Ovarian Insufficiency

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Familial primary ovarian insufficiency associated with an SYCE1 point mutation: defective meiosis elucidated in humanized mice

MHR Basic science of reproductive medicine ◽

10.1093/molehr/gaaa032 ◽

2020 ◽

Vol 26 (7) ◽

pp. 485-497

Author(s):

Diego Hernández-López ◽

Adriana Geisinger ◽

María Fernanda Trovero ◽

Federico F Santiñaque ◽

Mónica Brauer ◽

...

Keyword(s):

Mouse Model ◽

Central Element ◽

Transcript Level ◽

Primary Ovarian Insufficiency ◽

Primary Amenorrhea ◽

Causative Role ◽

Ovarian Insufficiency ◽

Humanized Mouse Model ◽

Chromosome Synapsis ◽

Human Mutation

Abstract More than 50% of cases of primary ovarian insufficiency (POI) and nonobstructive azoospermia in humans are classified as idiopathic infertility. Meiotic defects may relate to at least some of these cases. Mutations in genes coding for synaptonemal complex (SC) components have been identified in humans, and hypothesized to be causative for the observed infertile phenotype. Mutation SYCE1 c.721C>T (former c.613C>T)—a familial mutation reported in two sisters with primary amenorrhea—was the first such mutation found in an SC central element component-coding gene. Most fundamental mammalian oogenesis events occur during the embryonic phase, and eventual defects are identified many years later, thus leaving few possibilities to study the condition’s etiology and pathogenesis. Aiming to validate an approach to circumvent this difficulty, we have used the CRISPR/Cas9 technology to generate a mouse model with an SYCE1 c.721C>T equivalent genome alteration. We hereby present the characterization of the homozygous mutant mice phenotype, compared to their wild type and heterozygous littermates. Our results strongly support a causative role of this mutation for the POI phenotype in human patients, and the mechanisms involved would relate to defects in homologous chromosome synapsis. No SYCE1 protein was detected in homozygous mutants and Syce1 transcript level was highly diminished, suggesting transcript degradation as the basis of the infertility mechanism. This is the first report on the generation of a humanized mouse model line for the study of an infertility-related human mutation in an SC component-coding gene, thus representing a proof of principle.

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