EGF-FSH supplementation reduces apoptosis of pig granulosa cells in co-culture with cumulus-oocyte complexes

Palmerini, Maria Grazia; Nottola, Stefania Annarita; Tunjung, Woro Anidito Sri; Kadowaki, Akane; Bianchi, Serena; Cecconi, Sandra; Sato, Eimei; Macchiarelli, Guido

doi:10.1016/j.bbrc.2016.10.151

Cited by 27 publications

(15 citation statements)

References 40 publications

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“…Additionally, vitrification significantly decreased the expression of Dnmt1 messenger RNA (mRNA) in mouse MII oocytes [100,101]. However, in the absence of cryopreservation, the process of IVM may induce degeneration via apoptosis [102,103]. This pathway may be activated by stimuli of temperature, toxicants, and oxidative stress.…”

Section: Molecular Integrity Of Ivm Oocytes and Vitrificationmentioning

confidence: 99%

Vitrification of human immature oocytes before and after in vitro maturation: a review

Khalili

Shahedi

Ashourzadeh

et al. 2017

J Assist Reprod Genet

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The use of immature oocytes subjected to in vitro maturation (IVM) opens interesting perspectives for fertility preservation where ovarian reserves are damaged by pathologies or therapies, as in PCO/PCOS and cancer patients. Human oocyte cryopreservation may offer some advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation and postponing childbirth. It also eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In addition, a successful oocyte cryopreservation program could eliminate the need for donor and recipient menstrual cycle synchronization. Recent advances in vitrification technology have markedly improved the oocyte survival rate after warming, with fertilization and implantation rates comparable with those of fresh oocytes. Healthy live births can be achieved from the combination of IVM and vitrification, even if vitrification of in vivo matured oocytes is still more effective. Recently, attention is given to highlight whether vitrification procedures are more successful when performed before or after IVM, on immature GV-stage oocytes, or on in vitro matured MII-stage oocytes. In this review, we emphasize that, even if there are no differences in survival rates between oocytes vitrified prior to or post-IVM, reduced maturation rates of immature oocytes vitrified prior to IVM can be, at least in part, explained by underlying ultrastructural and biomolecular alterations.

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Section: Molecular Integrity Of Ivm Oocytes and Vitrificationmentioning

confidence: 99%

Vitrification of human immature oocytes before and after in vitro maturation: a review

Khalili

Shahedi

Ashourzadeh

et al. 2017

J Assist Reprod Genet

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“…Ampullar samples of the right FTs were processed for conventional [ 37 , 38 , 39 , 40 , 41 ] and variable pressure (VPSEM) SEM analysis. For each experimental group (Ctr, 4R, 8R), the isolated oviducts of their neighboring ovaries were fixed in 2.5% glutaraldehyde/0.1 M PBS at 4°C for at least 48 h. Selected sections of ampulla were processed either for conventional or VPSEM preparation.…”

Section: Methodsmentioning

confidence: 99%

Repeated hyperstimulation affects the ultrastructure of mouse fallopian tube epithelium

et al. 2020

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Controlled ovarian hyperstimulation (COH) is routinary used in assisted reproductive technologies (ARTs) to increase the yields of mature oocytes. The possibility that patients with a history of failures or poor-responders may develop side-effects following these treatments is still debated. Epidemiological studies reported controversial results about pregnancy outcome and the risk of developing gynecological cancers. By using a mouse model, here we compared the ultrastructural features of fallopian tubes (FTs) obtained from mice undergoing or not (control, CTR) four (4R) and eight (8R) rounds of gonadotropin stimulation. Although the morphological characteristics of oviductal layers seemed unaffected by repeated treatments, dose-response ultrastructural alterations in the ampulla appeared in the 4R group and even more in the 8R group. The targets were oviductal ciliated (CCs) and non-ciliated (NCCs) cells, which showed damaged mitochondria and glycogen accumulations in the cytoplasm. The drastic reduction of CCs, evident after 4R, was supported by the absence of cilia. After 8R, glycogen granules were significantly reduced and massive degeneration of mitochondria, which appeared swollen and/or vacuolated, occurred in NCCs. Moreover, disintegrated mitochondria were found at the periphery of mitophagic vacuoles with evident signs of cristolysis. The morphometric analysis evidenced a significant increase in the density and frequency of damaged mitochondria after 4R and 8R. The absence of cilia, necessary to sustain oviductal transport of oocytes, spermatozoa and embryos, may originate from either mitochondrial dysfunction or glycogen consumption. These results suggest that repeated COH treatments could induce alterations impairing fertilization and embryo transport toward the uterus.

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“…For SEM observations, IVC was performed on 18-mm sterile polylisinated coverslips placed in culture dishes. Next, adherent GCs were washed in PBS, fixed in 2.5% glutaraldehyde/PBS at 4°C for at least 48 h, and then subjected to SEM specimen preparation [ 21 , 22 , 23 , 24 , 25 , 26 ]. Briefly, fixed GCs were post-fixed with osmium tetroxide, washed in PBS, and dehydrated with increasing concentrations of alcohol.…”

Section: Methodsmentioning

confidence: 99%

Mancozeb impairs the ultrastructure of mouse granulosa cells in a dose-dependent manner

Palmerini

Belli

Nottola

et al. 2018

Journal of Reproduction and Development

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Mancozeb, an ethylene bis-dithiocarbamate, is widely used as a fungicide and exerts reproductive toxicity in vivo and in vitro in mouse oocytes by altering spindle morphology and impairing the ability to fertilize. Mancozeb also induces a premalignant status in mouse granulosa cells (GCs) cultured in vitro, as indicated by decreased p53 expression and tenuous oxidative stress. However, the presence and extent of ultrastructural alterations induced by mancozeb on GCs in vitro have not yet been reported. Using an in vitro model of reproductive toxicity, comprising parietal GCs from mouse antral follicles cultured with increasing concentrations of mancozeb (0.001–1 µg/ml), we sought to ascertain the in vitro ultrastructural cell toxicity by means of transmission (TEM) and scanning (SEM) electron microscopy. The results showed a dose-dependent toxicity of mancozeb on mouse GCs. Ultrastructural data showed intercellular contact alterations, nuclear membrane irregularities, and chromatin marginalization at lower concentrations, and showed chromatin condensation, membrane blebbing, and cytoplasmic vacuolization at higher concentrations. Morphometric analysis evidenced a reduction of mitochondrial length in GCs exposed to mancozeb 0.01−1 µg/ml and a dose-dependent increase of vacuole dimension. In conclusion, mancozeb induced dose-dependent toxicity against GCs in vitro, including ultrastructural signs of cell degeneration compatible with apoptosis, likely due to the toxic breakdown product ethylenethiourea. These alterations may represent a major cause of reduced/delayed/missed oocyte maturation in cases of infertility associated with exposure to pesticides.

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EGF-FSH supplementation reduces apoptosis of pig granulosa cells in co-culture with cumulus-oocyte complexes

Cited by 27 publications

References 40 publications

Vitrification of human immature oocytes before and after in vitro maturation: a review

Vitrification of human immature oocytes before and after in vitro maturation: a review

Repeated hyperstimulation affects the ultrastructure of mouse fallopian tube epithelium

Mancozeb impairs the ultrastructure of mouse granulosa cells in a dose-dependent manner

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