Modulation of P2Y2 receptors in bovine cumulus oocyte complexes: effects on intracellular calcium, zona hardening and developmental competence

Fonseca, Élisa; Mesquita, Patrı́cia; Marques, C.C.; Baptista, M.C.; Pimenta, Jorge; Matos, Joana; Soveral, Graça; Pereira, Manuel F. O. Seabra

doi:10.1007/s11302-020-09690-6

Cited by 4 publications

(8 citation statements)

References 34 publications

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“…Only COCs with at least three layers of compact cumulus cells and a homogeneous ooplasm were washed and selected for maturation according to the experimental design. Maturation was accomplished in an incubator at 38.8 • C, 5% CO 2 in humidified air for 22 or 30 h in a maturation medium composed of tissue culture medium 199 (TCM) with 10% of fetal bovine serum, 0.2 mM sodium pyruvate, 10 ng mL −1 of epidermal growth factor, and 10 µL mL −1 of gentamicin [40].…”

Section: Oocyte Collection and In Vitro Maturationmentioning

confidence: 99%

“…COCs and sperm were co-incubated for 20 h at 38.8 • C and 5% CO 2 in humidified air. Presumptive zygotes were then transferred to droplets of synthetic oviductal fluid (SOF) medium supplemented with BME and MEM amino acids, glutamine, glutathione, and BSA [40]. After 48 h of the insemination, the cleavage rate (cleaved embryos per total inseminated oocytes) was assessed, and cleaved embryos were maintained in SOF supplemented with BSA and 10% of fetal bovine serum (FBS).…”

Section: In Vitro Fertilization and Embryo Culturementioning

confidence: 99%

“…Embryos were cultured for 12 days [41,43] to assess the blastocyst development rate (at days 7, 9, and 12; D7 embryos per cleaved embryos) and hatched embryo rate (hatched embryos per D7 embryos) and their quality [43]. Day 7 embryos were classified as grade 1 (good quality), 2 (fair quality), and grade 3 (bad quality) [40,44].…”

Section: In Vitro Fertilization and Embryo Culturementioning

confidence: 99%

“…Total DNA and RNA were isolated from GCs using the High Pure PCR Template Preparation Kit (Roche, Basel, Switzerland) and PureLink™ RNA Mini Kit (Invitrogen™, Waltham, MA, USA), respectively, according to the manufacturer's instructions. Those protocols included the use of spin columns used to isolate high-quality total DNA and RNA, and DNase as treatment to remove genomic DNA from RNA [40]. After extraction, the samples were stored at −80 • C. The concentration and quality of DNA and RNA were determined using a NanoDrop™ One/OneC Spectrophotometer (ThermoFisher Scientific™, Waltham, MA, USA).…”

Section: Dna and Rna Extraction And Quantificationmentioning

confidence: 99%

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Anti-Aging Effect of Urolithin A on Bovine Oocytes In Vitro

Fonseca

Marques

Pimenta

et al. 2021

Animals

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Oxidative stress and mitochondrial dysfunction have been associated with the age-related decline of oocyte quality and strategies for their prevention are currently quested. Urolithin A (UA) is a natural metabolite with pro-apoptotic and antioxidant effects, capable of preventing the accumulation of dysfunctional mitochondria in different aged cells. UA has never been tested in bovine oocytes. Our aim was to study the effect of UA on the developmental potential of cumulus-oocyte-complexes (COCs) and granulosa cells’ (GCs) expression of important genes related to reproductive competence. Nuclear maturation progression, mitochondrial membrane potential (MMP) and developmental competence of physiologically mature (22 h) and in vitro aged oocytes (30 h of IVM) obtained from prepubertal and adult females, either supplemented with UA or not were assessed. Additionally, the amount of mRNA of several genes (NFE2L2, NQO1, and mt-DN5) and the number of mt-ND5 DNA copies were quantified in cultured GCs from prepubertal and adult females, either supplemented with UA or not. Our study confirmed the harmful effect of oocyte aging on the nuclear maturation progression, MMP, developmental competence and gene expression levels. UA treatment during in vitro maturation enhanced (p < 0.05) the maturation rate and subsequent developmental capacity of aged oocytes. A positive effect (p < 0.05) of UA on physiological maturation, MMP and embryonic development was also identified. UA also interfered on the expression profile of NFE2L2 and NQO1 genes in GCs cultures. Our findings demonstrate that UA supplementation is an effective way to prevent oocyte aging and improves the subsequent bovine embryonic development.

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Section: Oocyte Collection and In Vitro Maturationmentioning

confidence: 99%

Section: In Vitro Fertilization and Embryo Culturementioning

confidence: 99%

Section: In Vitro Fertilization and Embryo Culturementioning

confidence: 99%

Section: Dna and Rna Extraction And Quantificationmentioning

confidence: 99%

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Anti-Aging Effect of Urolithin A on Bovine Oocytes In Vitro

Fonseca

Marques

Pimenta

et al. 2021

Animals

Self Cite

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“…Then, oocytes with uniformly granular cytoplasm and surrounded by at least 3 layers of compact cumulus cells were selected for maturation [ 21 ]. The COCs were matured in tissue culture medium 199 (TCM199) with 10% serum, 10 ng mL −1 epidermal growth factor (EGF), 0.2 mM sodium pyruvate, and antibiotics, in an incubator at 38.8 °C with an atmosphere saturated with humidity and 5% CO 2 , for 22 h [ 22 ].…”

Section: Methodsmentioning

confidence: 99%

Effect of a Novel Hydroxybenzoic Acid Based Mitochondria Directed Antioxidant Molecule on Bovine Sperm Function and Embryo Production

Santos

Marques

Baptista

et al. 2022

Animals

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Sperm cells are particularly vulnerable to reactive oxygen species (ROS), impairing their fertilizing ability. Our objective was to study the effect of a novel mitochondrial-directed antioxidant, AntiOxBEN2, on bovine sperm function. This antioxidant was added to the semen capacitation medium (CAP), during the swim-up process, and to the fertilization medium (FERT) during the co-incubation of matured oocytes and capacitated spermatozoa, in concentrations of 0 (control), 1, and 10 µM. After the swim-up, sperm motility (CASA and visual analysis), vitality (eosin-nigrosin), mitochondrial membrane potential (JC1), intracellular ROS, adenosine triphosphate (ATP) levels, and basal metabolism (Seahorse Xfe96) were evaluated. Embryo development and quality were also assessed. Higher cleavage rates were obtained when 1 µM AntiOxBEN2 were added to CAP and FERT media (compared to control, p < 0.04). A positive effect of AntiOxBEN2 on intracellular ROS reduction (p = 0.01), on the increment of mitochondrial membrane potential (p ≤ 0.003) and, consequently, on the sperm quality was identified. However, the highest dose impaired progressive motility, ATP production, and the number of produced embryos. The results demonstrate a beneficial effect of AntiOxBEN2 (1 µM) on sperm capacitation and fertilization processes, thus improving embryonic development. This may constitute a putative novel therapeutic strategy to improve the outcomes of assisted reproductive techniques (ART).

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Calcium signaling in oocyte quality and functionality and its application

Chen,

Huang,

Dong

et al. 2024

Front. Endocrinol.

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Calcium (Ca2+) is a second messenger for many signal pathways, and changes in intracellular Ca2+ concentration ([Ca2+]i) are an important signaling mechanism in the oocyte maturation, activation, fertilization, function regulation of granulosa and cumulus cells and offspring development. Ca2+ oscillations occur during oocyte maturation and fertilization, which are maintained by Ca2+ stores and extracellular Ca2+ ([Ca2+]e). Abnormalities in Ca2+ signaling can affect the release of the first polar body, the first meiotic division, and chromosome and spindle morphology. Well-studied aspects of Ca2+ signaling in the oocyte are oocyte activation and fertilization. Oocyte activation, driven by sperm-specific phospholipase PLCζ, is initiated by concerted intracellular patterns of Ca2+ release, termed Ca2+ oscillations. Ca2+ oscillations persist for a long time during fertilization and are coordinately engaged by a variety of Ca2+ channels, pumps, regulatory proteins and their partners. Calcium signaling also regulates granulosa and cumulus cells’ function, which further affects oocyte maturation and fertilization outcome. Clinically, there are several physical and chemical options for treating fertilization failure through oocyte activation. Additionally, various exogenous compounds or drugs can cause ovarian dysfunction and female infertility by inducing abnormal Ca2+ signaling or Ca2+ dyshomeostasis in oocytes and granulosa cells. Therefore, the reproductive health risks caused by adverse stresses should arouse our attention. This review will systematically summarize the latest research progress on the aforementioned aspects and propose further research directions on calcium signaling in female reproduction.

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Modulation of P2Y2 receptors in bovine cumulus oocyte complexes: effects on intracellular calcium, zona hardening and developmental competence

Cited by 4 publications

References 34 publications

Anti-Aging Effect of Urolithin A on Bovine Oocytes In Vitro

Anti-Aging Effect of Urolithin A on Bovine Oocytes In Vitro

Effect of a Novel Hydroxybenzoic Acid Based Mitochondria Directed Antioxidant Molecule on Bovine Sperm Function and Embryo Production

Calcium signaling in oocyte quality and functionality and its application

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