Oxygen tension during in vitro culture of bovine embryos: Effect in production and expression of genes related to oxidative stress

Corrêa, G. A.; Rumpf, R.; Mundim, Tatiane Carmo Duarte; Franco, M. M.; Dode, M. A. N.

doi:10.1016/j.anireprosci.2007.02.002

Cited by 81 publications

(76 citation statements)

References 52 publications

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“…However, better embryo development after IVM under lower oxygen tension was also reported (Hashimoto et al, 2000), which may reflect other effects of medium composition and culture conditions. Some studies (Papis et al, 2007;Corrêa et al, 2008) assumed that higher oxygen tensions during IVC resulted in ROS production that was detrimental for embryo development. However, it was previously demonstrated that physiological concentrations of ROS may, therefore, play a key role in the process of oocyte maturation (Dalvit et al, 2005).…”

Section: Tablementioning

confidence: 99%

The effect of interaction between macromolecule supplement and oxygen tension on bovine oocytes and embryos culturedin vitro

Mingoti

Castro

Méo

et al. 2009

Zygote

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SummaryAiming to improve in vitro production of bovine embryos and to obtain supplements to replace serum for in vitro maturation (IVM), this study evaluated the effects of macromolecular supplementation of IMV medium (bovine serum albumin -BSA, polyvinyl alcohol -PVA, polyvinyl pyrrolidone -PVP, Ficoll, KnockoutSR, or fetal calf serum -FCS) and oxygen tension [5% CO 2 in air (20% O 2 ) or 5% CO 2 , 5% O 2 and 90% N 2 (5% O 2 )] on oocyte maturation and embryo development. Nuclear progression to germinal vesicle breakdown, metaphase I and metaphase II stages were evaluated and overall results revealed that undefined (FCS) and semi-defined (BSA) media gave better results at 20% O 2 and defined media (PVA, PVP and Ficoll) at 5% O 2 . Independent of macromolecule supplement, IVM at 20% O 2 was considered optimal for nuclear maturation. To evaluate embryo development, oocytes matured in the previously described conditions were fertilized and cultured at the same oxygen tension used for IVM and assessed for cleavage (43.0 to 74.8%) and development to morulae (16.4 to 33.8%), blastocyst (7.7 to 52.9%) and hatched blastocyst (9.6 to 48.1%). Apart from oxygen tension, all treatments, except Knockout (22.7%), gave similar results for blastocyst development (26.5 to 38.7%). Independently of macromolecule supplement, higher development rates were obtained in an oxygen tension of 20% O 2 (67.4% cleavage, 29.2% morulae, 40.8% blastocyst and 34.0% hatched blastocyst) when compared with 5% O 2 (52.5, 21.8, 18.2 and 15.6%, respectively). This study indicates that BSA, PVA, PVP and Ficoll can replace serum during IVM and that the optimal atmospheric condition for in vitro production of bovine embryos is 5% CO 2 and 20% O 2 .

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Section: Tablementioning

confidence: 99%

The effect of interaction between macromolecule supplement and oxygen tension on bovine oocytes and embryos culturedin vitro

Mingoti

Castro

Méo

et al. 2009

Zygote

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“…The in vitro culture procedures were based on methods described previously (Adona et al, 2008;Corrêa et al, 2008). Ovaries obtained from slaughterhouse cows were transported to the laboratory in saline solution (NaCl 0.9%) supplemented with 100 UI mL -1 penicillin G and 100 μg mL -1 streptomycin sulfate at 35°C within 2 h. Cumulus-oocyte complexes (COCs) were aspirated from 2-to 6-mm follicles with an 18 gauge needle and a 10 mL syringe and the aspirated follicular fluid was pooled in a 15 mL conical tube.…”

Section: In Vitro Maturation Fertilization and Embryo Culture Procedmentioning

confidence: 99%

“…It has been reported that FSH and LH added to maturation medium have a beneficial effect on the competence of oocytes to develop in vitro to the blastocyst stage (Papis et al, 2007;Adona et al, 2008;Corrêa et al, 2008). Although producing controversial results (Holm et al, 2002), FCS is successfully used as protein supplement in oocyte maturation medium (Sargirkaya et al, 2007;Adona et al, 2008;Corrêa et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

“…The basic maturation media used in bovine embryo culture systems are generally composed of TCM-199 medium supplemented with a combination of FSH, LH and fetal calf serum (FCS) (Sargirkaya et al, 2007;Adona et al, 2008;Corrêa et al, 2008). It has been reported that FSH and LH added to maturation medium have a beneficial effect on the competence of oocytes to develop in vitro to the blastocyst stage (Papis et al, 2007;Adona et al, 2008;Corrêa et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

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Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos

Takada

Martins

Mingoti

et al. 2010

Sci. agric. (Piracicaba, Braz.)

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Melatonin (MEL) acts as a powerful scavenger of free radicals and direct gonadal responses to melatonin have been reported in the literature. Few studies, however, have evaluated the effect of MEL during in vitro maturation (IVM) on bovine embryos. This study tested the addition of MEL to maturation medium (MM) with no gonadotropins on nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos. Cumulus-oocyte complexes were aspirated from abattoir ovaries and cultured in MM (TCM-199 medium supplemented with 10% fetal calf serum -FCS) at 39°C and 5% CO 2 in air. After 24 hours of culture in MM with 0.5 μg mL -1 FSH and 5.0 μg mL -1 LH; 10 -9 M MEL) or 10 -9 M MEL, 0.5 μg mL -1 FSH and 5.0 μg mL -1 LH, the oocytes were stained with Hoechst 33342 to evaluate nuclear maturation rate. After in vitro fertilization and embryo culture, development rates were evaluated and the blastocysts were assessed for DNA damage by Comet assay. There was no effect of melatonin added to the MM, alone or in combination with gonadotropins, on nuclear maturation, cleavage and blastocyst rates. These rates ranged between 88% to 90%, 85% to 88% and 42% to 46%, respectively. The extent of DNA damage in embryos was also not affected by MEL supplementation during IVM. The addition of 10 -9 M MEL to the MM failed to improve nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos, but was able to properly substitute for gonadotropins during IVM. Key words: comet assay, embryos, gonadotropins, in vitro fertilization, melatonin Melatonina no meio de maturação não melhorou as taxas de maturação dos ovócitos, de desenvolvimento embrionário e a fragmentação do DNA dos embriões bovinos RESUMO: Melatonin (MEL) atua como um potente redutor de radicais livres. Efeito direto da MEL na função gonadal também foi observado. Existem poucos estudos relacionados ao efeito da MEL durante a maturação no desenvolvimento embrionário in vitro. Avaliou-se a adição de MEL no meio de maturação (sem gonadotrofinas) nas taxas de maturação nuclear e de desenvolvimento embrionário e na incidência de fragmentação do DNA nos embriões produzidos in vitro. Complexos cumulus-ovócitos foram aspirados de ovários provenientes de matadouros e cultivados em meio de maturação (Meio TCM-199 suplementado com 10% de soro fetal bovino) a 39°C e 5% CO 2 em ar. Após 24 horas de cultivo em meio de maturação com 0,5 μg mL -1 de FSH e 5,0 μg mL -1 de LH; 10 -9 M de MEL ou 10 -9 M de MEL, 0,5 μg mL -1 de FSH e 5,0 μg mL -1 de LH, os ovócitos foram corados com Hoechst 33342 para avaliação da taxa de maturação nuclear. A fragmentação do DNA dos blastocistos foi avaliada pela técnica do Cometa. Não houve efeito da MEL adicionada ao meio de maturação, com ou sem gonadotrofinas, nas taxas de maturação nuclear, clivagem e blastocistos. Os percentuais variaram entre 88% a 90%, 85% a 88% e 42% a 46%, respectivamente. O grau de fragmentação do DNA também não foi afetado pela adição de MEL ao meio de matur...

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“…ATF4 induces a set of genes involved in amino acid metabolism, antioxidant stress response, and protein secretion, all of which aid in homeostasis [40]. The MnSOD gene encodes manganese superoxide dismutase, which exists in mitochondria for the conversion of superoxide to hydrogen peroxide, and plays important roles in oxidative stress response [41]. The Gadd45g family of stress-inducible acidic nuclear proteins is implicated in apoptosis and DNA repair [42,43], and the significant upregulation of the three gene members indicated that 3 h after warming, eight-cell embryos from obese mice bore more stress induced by obesity compared to their counterparts.…”

Section: The Effect Of Obesity On Vitrificationmentioning

confidence: 99%

Maternal obesity in mice not only affects fresh embryo quality but also aggravates injury due to vitrification

Huang

Yang

et al. 2016

J Assist Reprod Genet

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Purpose The aims of the present study are to identify the mechanism(s) whereby obesity impairs fresh embryos and to clarify the effects of vitrification on lipid droplet content within embryos from maternally obese mice. Methods The diet-induced obesity mouse model was established, and the zygotes were captured and cultured to day 3. The eight-cell embryos were selected and divided into fresh and vitrified groups. The blastocysts derived from fresh embryos were used as a control. The expression profiles of endoplasmic reticulum (ER) stress genes (Atf4, Grp78, and Hsp70) and other genes (MnSOD, p53, Gadd45g, caspase-3, IGF-II, ZO-1, and E-cadherin) on day-3 fresh and postwarming eight-cell embryos from obese and control groups were determined. For day-5 fresh blastocysts and blastocysts previously vitrified on day 3, the expression profiles for all of the above genes were also determined. Results For the fresh group, obesity significantly upregulated Hsp70, p53, IGF-II, and ZO-1 expression in embryos on day 3 and notably upregulated Atf4, MnSOD, Gadd45g, caspase-3, ZO-1, and E-cadherin expression in blastocysts on day 5. For vitrified ones, obesity significantly upregulated Atf4, MnSOD, and Gadd45g expression in embryos on day 3 and notably upregulated Hsp70 expression and downregulated MnSOD in day 5 blastocysts previously vitrified on day 3. Conclusions Obesity impairs fresh embryos and aggravates embryonic vitrification injury at a molecular level.

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Oxygen tension during in vitro culture of bovine embryos: Effect in production and expression of genes related to oxidative stress

Cited by 81 publications

References 52 publications

The effect of interaction between macromolecule supplement and oxygen tension on bovine oocytes and embryos culturedin vitro

The effect of interaction between macromolecule supplement and oxygen tension on bovine oocytes and embryos culturedin vitro

Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos

Maternal obesity in mice not only affects fresh embryo quality but also aggravates injury due to vitrification

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