Quercetin influences in vitro maturation, apoptosis and metabolically active mitochondria of goat oocytes

SummaryThe present study aimed to investigate the effect of quercetin as an alternative antioxidant to cysteamine on in vitro maturation. Oocytes were collected from goat ovaries, destined for in vitro maturation and distributed into three groups: CIS group, oocytes were immersed in MIV base medium; in Groups Q4 and Q8, oocytes were immersed in the medium of the CIS group, adding 4 μM or 8 μM of quercetin, respectively, and cultured for 24 h at 38.5°C with 5% CO2. The CIS and Q4 groups presented the same percentage of expanded cumulus cells, but the per cent in the Q8 group was significantly lower than that of the other groups (P<0.05). The oocyte retraction rate in the Q8 group was higher (P<0.05) than in the CIS and Q4 groups. Treatment with 8 μM of quercetin presented a lower proportion of expanded oocytes than the CIS group and 4 μM of quercetin (P<0.05). The percentage of MII oocytes was higher in the Q4 group than in the CIS group (P<0.05), but the percentages in the CIS and Q8 groups were similar. The rate of apoptosis was higher in the CIS group than in the other groups (P<0.05). In addition, oocytes matured with 4 μM quercetin showed higher mitochondrial activity than matured oocytes in the CIS and Q8 groups (P<0.05). In conclusion, 4 μM of quercetin can be used as an alternative to cysteamine in the in vitro maturation of goat oocytes.

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In vitro maturation impacts cumulus–oocyte complex metabolism and stress in cattle

Reproduction ◽

10.1530/rep-17-0134 ◽

2017 ◽

Vol 154 (6) ◽

pp. 881-893 ◽

Cited By ~ 9

Author(s):

Maite del Collado ◽

Juliano C da Silveira ◽

Marcelo L F Oliveira ◽

Bárbara M S M Alves ◽

Rosineide C Simas ◽

...

Keyword(s):

In Vitro Maturation ◽

Neutral Lipids ◽

Cumulus Cells ◽

Acid Synthesis ◽

Cellular Stress Response ◽

Mitochondrial Activity ◽

Cumulus Oocyte Complex ◽

Glycolysis Pathway

The influence of in vitro maturation (IVM) in oocytes is still not totally understood. The aim of this study was to determine the influence of IVM on the metabolism and homeostasis of bovine cumulus-oocyte complexes. In the present study, we demonstrated that IVM leads to accumulation of neutral lipids associated with differential levels of the mono-, di- and triacylglycerols in both cumulus cells and oocytes. We observed that in vitro-matured oocytes exhibited decreased glutathione and reactive oxygen species levels and a lower ATP/ADP ratio when compared to in vivo-matured oocytes, with no significant differences in metabolism and stress-related mRNA or miRNA levels. Moreover, in addition to an increase in lipids in in vitro-matured cumulus cells, fatty acid synthesis and accumulation as well as glycolysis pathway genes were upregulated, whereas those affiliated with the β-oxidation pathway were decreased. Our gene expression data in cumulus cells suggest the disruption of endoplasmic reticulum stress, apoptosis and cellular stress response pathways during IVM. Furthermore, a total of 19 miRNAs were significantly altered by the maturation process in cumulus cells. These results indicate some new negative influences of the in vitro system in cumulus-oocyte complexes, demonstrating the occurrence of functional disruption in lipid metabolism and stress pathways and showing evidences suggesting the occurrence of altered mitochondrial activity and energy metabolism during IVM, with a massive dysregulation of the corresponding transcripts in the surrounding cumulus cells.

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Improved human oocyte in vitro maturation in co-culture with cumulus cells from mature oocytes in the same patient: a prospective study

10.26226/morressier.573c1514d462b80296c98d07 ◽

2016 ◽

Author(s):

Irma Virant - Klun

Keyword(s):

Prospective Study ◽

In Vitro Maturation ◽

Cumulus Cells ◽

Human Oocyte ◽

A Prospective Study

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46 Effect of the addition of α-tocopherol to in vitro maturation media of bovine oocytes

Journal of Animal Science ◽

10.1093/jas/skz397.005 ◽

2020 ◽

Vol 98 (Supplement_2) ◽

pp. 2-3

Author(s):

Theisy P Acosta Pérez

Keyword(s):

In Vitro Maturation ◽

Cumulus Cells ◽

Control Group ◽

Chi Square ◽

Cumulus Expansion ◽

Minimum Concentration ◽

Maturation Rate ◽

Chi Square Test ◽

System Data

Abstract α-tocopherol is known to be a powerful antioxidant, in this regard, it was added to bovine oocyte in vitro maturation media to evaluate its effect on oocyte maturation. Oocytes (n = 624) aspirated from ovaries of slaughtered cows were classified by quality and divided in four categories according to cytoplasm appearance and cumulus cells layers. Oocytes were washed in TCM-199 supplemented with fetal bovine serum (FBS) and FSH, then distributed in maturation media (TCM-199 supplemented with FBS, FSH and gentamicin). Three experimental groups of α-tocopherol (50, 100 and 200 mM) and a control group without α-tocopherol were used. Maturation was carried 22 h at 38.5°C in a 5% CO2 atmosphere. Oocytes were examined to determine cumulus expansion as categorical data (expansion or no expansion), as well as cumulus expansion Index (CEI). For CEI determination oocytes were graded 0 to 4, being 0 those with null expansion and 4 those with a noticeable cell expansion, then the number of oocytes were multiplied by the grade given and a sum of the totals was obtained, the new total was divided by the total of oocytes in the group and the result obtained corresponded to the CEI of the group. Results were analyzed with Chi Square test (for maturation rates) and an ANOVA (for the CEI) using the SAS system, data are presented as mean ± standard error. There was no statistical difference between control and α-tocopherol groups (P >0.05). Numerically, the control group showed a higher maturation rate (100%) and obtained a higher CEI (2.44±0.20), followed by the 50 mM group (98.16%; 2.39±0.13), the groups 200 mM (97.40%; 2.00±0.14) and 100 mM (96.25%; 2.06±0.24) were the lowest. The addition of the minimum concentration (50 mM) of α-tocopherol to the maturation media could improve maturation rates without exposing oocytes to toxic effects.

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Growth factors improve mouse oocyte developmental potential via increased MAPK and MTOR signaling activities in cumulus cells during in vitro maturation

Fertility and Sterility ◽

10.1016/j.fertnstert.2018.07.882 ◽

2018 ◽

Vol 110 (4) ◽

pp. e313 ◽

Cited By ~ 1

Author(s):

J.C. Becker ◽

R. Pasquariello ◽

S.K. Rajput ◽

W.B. Schoolcraft ◽

R.L. Krisher ◽

...

Keyword(s):

Growth Factors ◽

In Vitro Maturation ◽

Cumulus Cells ◽

Mtor Signaling ◽

Mouse Oocyte ◽

Developmental Potential

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Influence of cumulus cells on in vitro maturation and fertilization of equine oocytes

Theriogenology ◽

10.1016/0093-691x(96)84736-4 ◽

1996 ◽

Vol 45 (1) ◽

pp. 263 ◽

Cited By ~ 3

Author(s):

M.V. Marcos ◽

A.R. Spell ◽

M.D. Butine ◽

M.J. Arns

Keyword(s):

In Vitro Maturation ◽

Cumulus Cells

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Uptake and Metabolism of Tritiated 17ß-Oestradiol and Oestrone by Rabbit Uterine Tissue in vitro

Australian Journal of Biological Sciences ◽

10.1071/bi9740137 ◽

1974 ◽

Vol 27 (2) ◽

pp. 137 ◽

Cited By ~ 1

Author(s):

RG Gabb ◽

GM Stone

Keyword(s):

Nuclear Receptor ◽

Mode Of Action ◽

Nuclear Extract ◽

Lower Proportion ◽

The Other ◽

Uterine Tissue ◽

Uptake And Metabolism

To determine whether the established capability of rabbit uterine tissue to interconvert 17 p-oestradiol and oestrone might have some effect on the mode of action of the oestrogens in this organ, the in vitro interconversion of [3H]-17p-oestradiol and [3H]oestrone by rabbit endometrial and myometrial tissue was investigated and the identity of radiometabolites in 'soluble, 'mitochondrial-microsomal' a'nd 'nuclear' preparations was studied. Both endometrial and myometrial tissue were found to be capable of oxidoreduction of the oestrogens, the equilibrium of the reaction favouring the reduction of oestrone. Irrespective of the tissue--steroid combination studied, the greater part of the radioactivity in all fractions was associated with 17p-oestradiol. The relative proportions of [3Hl-17p-oestradiol and [3H]oestrone varied between fractions, the nuclear preparation consistently showing a lower proportion of oestrone than the other fractions. Sephade<c fractionation of a 0'4M KCl 'nuclear extract' revealed that proportionately less oestrone than 17p-oestradiol was bound to the nuclear 'receptor'. These findings provide further evidence for 17p-oestradiol being the ovarian oestrogen which is active in the uterus, and suggest a role for uterine oxidoreduction of oestrogens in the control exercised over this organ by these steroids.

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Transcriptional, Mitochondrial Activity, and Viability of Egyptian Buffalo’s Granulosa Cells In Vitro Cultured under Heat Elevation

World s Veterinary Journal ◽

10.54203/scil.2021.wvj25 ◽

2021 ◽

Vol 11 (2) ◽

pp. 193-201

Author(s):

Nasser Ghanem ◽

Marwa Said Faheem ◽

Romysa Samy ◽

Ashraf Hesham Barkawi

Keyword(s):

Heat Stress ◽

Granulosa Cells ◽

Transcriptional Activity ◽

The Other ◽

Control Group ◽

Mitochondrial Activity ◽

Fluorescent Intensity ◽

Stressed Group ◽

Other Hand

It is documented that heat stress caused impairment on the reproductive performance of dairy animals. However, there are few reports that have focused on the molecular and intracellular responses of in vitro cultured buffalo granulosa cells during heat elevation. The present study was conducted to investigate the effect of heat elevation during in vitro culture of buffalo granulosa cells on their viability, quality, mitochondrial activity, and transcriptional activity. Granulosa cells were harvested after aspiration of cumulus-oocytes complexes that were collected from abattoir ovaries. The granulosa cells were cultured in vitro either at a normal physiological temperature suitable for oocyte maturation and embryo development (38.5°C) or exposed to the elevated temperature of 40.5°C on day 3 of culture (the first two days were for confluence) for two hours of culture then continued at 38.5°C up to day 7 of culture. The viability of granulosa cells was measured using trypan blue and quality was estimated by measuring the level of intracellular reactive oxygen species (ROS) on day 7. Moreover, metabolic activity was performed by measuring the fluorescent intensity of mitochondria. Moreover, transcriptional activity was done by profiling four selected candidate genes using quantitative real-time PCR. The results indicated that the granulosa cells viability rate significantly decreased in the heat stress group (25.1 ± 3.7), compared to the control group (36.6 ± 5.3) on confluence day (day 3). In addition, the viability rate on the last day of culture (day 7) decreased in heat stress, compared to control (83.7 ± 4.5 and 97.4 ± 0.4, respectively). On the other hand, there was a nonsignificant difference in ROS profile between the control (21.7*104 ± 1.3) and the heat-stressed group (15.7 ± 0.7) on day 7 of culture. However, the mitochondrial fluorescent intensity was higher in the control (21.9 ± 1.9) than in the heat-stressed group (15.4 ± 0.8) on day 7 of culture. The expression of cellular defense (HSF1) and apoptosis-inducing gene (P53) were significantly up-regulated in granulosa cells exposed to heat elevation, compared to the control group. On the other hand, the steroidogenesis-regulating gene (StAR) was down-regulated in granulosa cells cultured under heat shock, compared to the control group. In conclusion, heat stress reduced the viability of granulosa cells by inducing the expression of an apoptosis-related gene (P53) and compromised expression of genes regulating the steroid biosynthesis, which resulted in up-regulation of cell defense gene (HSF1) in an attempt to ameliorate the deleterious effect of heat stress on the biological activity of the granulosa cells.

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