Toll‐like receptor4 as a modulator of fertilization and subsequent pre‐implantation development following in vitro maturation in mice

Hosseini, Sara; Dehghani-Mohammadabadi, Maryam; Novin, Marefat Ghafarri; Hoseini, Mostafa Haji Molla; Arefian, Ehsan; Yeganeh, Samira Mohammadi; Salehi, Mohammad

doi:10.1111/aji.12720

Cited by 17 publications

(18 citation statements)

References 41 publications

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“…Therefore duration time was gradually increased to 22–24 hr, which the highest maturation rate was seen in the oocytes. Thus in this study likewise the other reports, (Hosseini et al, ; Jafarzadeh et al, ; Junk, Dharmarajan, & Yovich, ; Zare et al, ), GV oocytes were incubated for 22–24 hr during IVM. After this time, the expansion rate was evaluated before any manipulations, and then the oocytes were detached from cumulus cells by mechanical pipetting for evaluation of nuclear maturation.…”

Section: Methodssupporting

confidence: 75%

Thymoquinone reduces intracytoplasmic oxidative stress and improves epigenetic modification in polycystic ovary syndrome mice oocytes, during in‐vitro maturation

Eini

Bidadkosh

Nazarian

et al. 2019

Molecular Reproduction Devel

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Although in‐vitro maturation (IVM) of oocytes has been presented as an alternative treatment to traditional stimulated in‐vitro fertilization, the culture condition can be improved by natural antioxidants. Thus, we investigated the protective effect of Thymoquinone (TQ) during IVM in the polycystic ovary syndrome (PCOS) mice model. The induction of PCOS was made by dehydroepiandrosterone via subcutaneous injection, in prepubertal female B6D2F1‐mice. After 21 days later, germinal vesicle (GV)‐stage‐oocytes were extracted and incubated in IVM media containing 0, 1.0, 10.0, and 100.0 μM of TQ. To assess fertilization and blastulation rates, after 22–24 hr, the treated oocytes were fertilized in‐vitro with epididymal spermatozoa. Some other oocytes were evaluated for maturation, epigenetic, and oxidative stress markers. Similarly, the mRNA expression of epigenetic enzymes genes (Dnmt1 and Hdac1), three maternally derived genes (Mapk, CyclinB, and Cdk1) and apoptosis‐related genes (Bax and Bcl2) were assessed. Our results showed that the maturation, fertilization, and blastulation rates were significantly higher in the 10.0 μM TQ‐treated group compared with the untreated group and likewise with in‐vivo matured oocytes. The Bax expression was reduced in 10.0 μM TQ matured oocytes, but Bcl2, Dnmt1, Hdac1, Cdk1, and Mapk were upregulated in this group compared to other groups. Furthermore, dimethylation of histone‐3 at lysine‐9 (H3K9m2) and DNA methylation were significantly increased whereas H4K12 acetylation (H4K12ac) was decreased in the 10.0 μM TQ‐treated group in comparison with control and in‐vivo matured oocytes. Therefore, our results are suggesting that 10.0 μM TQ may enhance the developmental competence of PCOS oocytes via the modulation of oxidative stress and epigenetic alterations.

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Section: Methodssupporting

confidence: 75%

Thymoquinone reduces intracytoplasmic oxidative stress and improves epigenetic modification in polycystic ovary syndrome mice oocytes, during in‐vitro maturation

Eini

Bidadkosh

Nazarian

et al. 2019

Molecular Reproduction Devel

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“…CM, conditioned media; MSC, mesenchymal stromal cell; PCOS, polycystic ovary syndrome; SEM, standard error of mean patients with PCOS, 30,38 those who suffer from repeated IVF-embryo transfer failure, as a result of defective oocyte or embryo qualities, 30 and those who do not respond to gonadotropin administration. Representative images (A) and quantitative data (B) for intracellular glutathione (GSH) levels, as cytoplasmic maturation marker, in the in vitro matured oocytes from PCOS and normal mice.…”

Section: Discussionmentioning

confidence: 99%

“…30 Ovaries were excised 48 hours later and placed in the prewarmed 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid tissue cell culture medium (TCM199) supplemented with 10% FBS. 30 Ovaries were excised 48 hours later and placed in the prewarmed 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid tissue cell culture medium (TCM199) supplemented with 10% FBS.…”

Section: Oocytes Collectionmentioning

confidence: 99%

“…patients with PCOS, 30,38 those who suffer from repeated IVF-embryo transfer failure, as a result of defective oocyte or embryo qualities, 30 and those who do not respond to gonadotropin administration. 3 In the current study, we have shown that when hBM-MSC-CM was used to enrich the conventional IVM medium, TCM199; it could improve meiotic and developmental competence of PCOS oocytes.…”

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confidence: 99%

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Improvement of oocyte in vitro maturation from mice with polycystic ovary syndrome by human mesenchymal stromal cell–conditioned media

Jafarzadeh

Nazarian

Novin

et al. 2018

J of Cellular Biochemistry

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The outcome of in vitro maturation (IVM) in the patients with polycystic ovary syndrome (PCOS) is poor. Abnormal intraovarian paracrine interplay alters microenvironment for oocyte development through folliculogenesis and decreases developmental competence of oocytes in patients with PCOS. Mesenchymal stromal cells (MSCs) secrete a variety of cytokines and growth factors that could promote oocyte maturation in vitro. Thus, in the current study we aimed to evaluate the effect of human bone marrow MSC-conditioned media (hBM-MSC-CM), as a supplement, to enrich IVM medium for PCOS germinal vesicles (GVs). For this purpose, oocytes at GV and metaphase II (MII) stages were harvested from PCOS mice. The GVs were randomly divided into four groups and incubated for 24 hours in an IVM medium (TCM199, as the control group) or TCM199 supplemented by 25%, 50%, and 75% of hBM-MSC-CM (PCOS-CM25, PCOS-CM50, and PCOS-CM75 groups, respectively) so as to evaluate which dose(s) could enhance maturation rate of the GVs and their subsequent in vitro fertilization (IVF) outcome. Furthermore, MII oocytes and their subsequent IVF outcome were considered as the in vivo matured (PCOS-IVO) group. The data showed that supplementation of IVM medium with 50% hBM-MSC-CM significantly increased cytoplasmic and nuclear maturation of the GVs (P < 0.001), and also fertilization and two-cell rate (P < 0.001) and blastocyst formation (P < 0.01) of in vitro matured oocytes from mice with PCOS. Overall, higher oocyte maturation and fertilization outcome in PCOS-CM50 group proposed that enrichment of IVM medium with hBM-MSC-CM could be considered as a promising approach to improve IVM of PCOS oocytes.

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“…Complementary DNA (cDNA) synthesis and quantitative real‐time PCR (qRT‐PCR) were performed according to a protocol by Hosseini et al (). In order to produce cDNA, 7 µl of nuclease‐free water and 1 µl poly N were added to 4 µl of the RNA sample.…”

Section: Methodsmentioning

confidence: 99%

DNA integrity and methylation changes of mouse spermatozoa following prolonged incubation

et al. 2019

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Sperm quality can be affected by different factors including the length of incubation time between sperm preparation and intracytoplasmic sperm injection. Here, we have evaluated the level of DNA methylation and expressions of related genes in mice spermatozoa. The spermatozoa were divided into three groups: fresh, spermatozoa incubated at room temperature (RT) and 37°C for 24 hr. The sperm chromatin structure assay was used to determine the DNA fragmentation index (DFI), and DNA methylation was analysed by flow cytometry. The expression levels of DNA methylation‐related genes were determined by quantitative real‐time PCR (qRT‐PCR). According to the results, we observed significantly higher sperm progressive motility and viability in the group incubated at RT compared to the spermatozoa incubated at 37°C (p < 0.05). Spermatozoa incubated at 37°C had a higher DFI compared to the other groups (p < 0.05), but the DNA methylation level significantly decreased (p < 0.05). qRT‐PCR analysis showed increased Dnmt‐1 expression in spermatozoa after 24‐hr incubation at 37°C. However, there were significantly higher expression levels of Dnmt‐3l, Dnmt‐3a and Dnmt‐3b after incubation at both RT and 37°C compared to the fresh group (p < 0.05). The 24‐hr incubation period affected both sperm DNA methylation and integrity. This study indicated that incubation at RT resulted in better sperm quality.

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Toll‐like receptor4 as a modulator of fertilization and subsequent pre‐implantation development following in vitro maturation in mice

Abstract: Low expression level of Tlr4 following IVM and higher fertilization rate through TLR4 receptor activation with LPS proposed that alteration in TLR4 expression and subsequent cytokine section could be a possible cause of low fertilization rate in IVM-derived oocytes.

Cited by 17 publications

References 41 publications

Thymoquinone reduces intracytoplasmic oxidative stress and improves epigenetic modification in polycystic ovary syndrome mice oocytes, during in‐vitro maturation

Thymoquinone reduces intracytoplasmic oxidative stress and improves epigenetic modification in polycystic ovary syndrome mice oocytes, during in‐vitro maturation

Improvement of oocyte in vitro maturation from mice with polycystic ovary syndrome by human mesenchymal stromal cell–conditioned media

DNA integrity and methylation changes of mouse spermatozoa following prolonged incubation

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