Parthenogenetic activation of bovine oocytes using single and combined strontium, ionomycin and 6-dimethylaminopurine treatments

Méo, Simone Cristina; Yamazaki, W.; Ferreira, Christina R.; Perecin, Felipe; Saraiva, Naiara Zoccal; Leal, Cláudia Lima Verde; Garcia, Joaquim Mansano

doi:10.1017/s0967199407004285

Cited by 25 publications

(13 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The low blastocyst rates obtained when fertilization was performed at 28 h of IVM might be explained by the fact that fertilization does not occur immediately after insemination. According to Méo et al (2007), the pronuclei first appear 12 h after insemination. Thus, oocytes inseminated later (28 h) would have missed the optimal moment for fertilization due to aging.…”

Section: Resultsmentioning

confidence: 99%

Effect of follicular fluid supplementation during in vitro maturation on total cell number in bovine blastocysts produced in vitro

et al. 2014

View full text Add to dashboard Cite

-This study evaluated the influence of follicular fluid (FF) added to the maturation medium on the quality of bovine embryos produced in vitro. In the first experiment, oocytes were matured in media containing five different FF concentrations with different maturation times and classified according to meiotic progression and migration of cortical granules. In the second experiment, oocytes matured in the same media were fertilized at three different maturation times; thereafter, cleavage and blastocyst rates were evaluated. In the third experiment, oocytes were matured in media containing three different FF concentrations at two different maturation times, and embryo quality, inferred by the ratio of inner cell mass and trophectoderm cells compared with total cell number, was evaluated. Higher FF concentration (75 -100% FF) slowed meiotic progression and CG migration (control -78.13% vs. treated -52.58% and control -52.7% vs. treated -11.59%, respectively, at 24 h of maturation). Also, FF at concentration of 75% or 100% had a negative influence on cleavage and blastocyst rates (control -90.13% vs. treated -82.64% and control -35.73% vs. treated -11.57%, respectively, at 24 h of maturation). The 50% FF resulted in embryos with increased inner cell mass numbers (control -29.91 vs. treated -35.49, at 24 h of maturation) and total cell numbers (control -109.53 vs. treated -120.67, at 26 h of maturation). Even though higher concentration of FF added to the maturation medium reduced embryonic development rates, in lower concentrations, FF slowed the meiotic progression and migration of CG and contributed to increases in inner cell mass number. Thus, FF added to the maturation medium enhances the number of cells in bovine embryos produced in vitro, especially for inner cell mass.

show abstract

Section: Resultsmentioning

confidence: 99%

Effect of follicular fluid supplementation during in vitro maturation on total cell number in bovine blastocysts produced in vitro

et al. 2014

View full text Add to dashboard Cite

show abstract

“…The Ca 2+ ionophore ionomycin has also been shown to induce [Ca 2+ ] i elevations in occytes of many species. According to the literature, the parameters of ionomycin treatment used to activate oocytes of various species vary significantly: concentrations ranging from 5 μM to 5 mM [33] and treatment duration of 1 min to 40 min have also been reported [34]. The influence of ionomycin on cat oocytes has never been described; therefore optimizing the concentration and incubation time is critical for successful oocyte activation.…”

Section: Discussionmentioning

confidence: 99%

Analysis of cat oocyte activation methods for the generation of feline disease models by nuclear transfer

Wang

Swanson

Herrick

et al. 2009

Reprod Biol Endocrinol

View full text Add to dashboard Cite

BackgroundSomatic cell nuclear transfer in cats offers a useful tool for the generation of valuable research models. However, low birth rates after nuclear transfer hamper exploitation of the full potential of the technology. Poor embryo development after activation of the reconstructed oocytes seems to be responsible, at least in part, for the low efficiency. The objective of this study was to characterize the response of cat oocytes to various stimuli in order to fine-tune existing and possibly develop new activation methods for the generation of cat disease models by somatic cell nuclear transfer.MethodsFirst, changes in the intracellular free calcium concentration [Ca2+]i in the oocytes induced by a number of artificial stimuli were characterized. The stimuli included electroporation, ethanol, ionomycin, thimerosal, strontium-chloride and sodium (Na+)-free medium. The potential of the most promising treatments (with or without subsequent incubation in the presence of cycloheximide and cytochalasin B) to stimulate oocyte activation and support development of the resultant parthenogenetic embryos was then evaluated. Finally, the most effective methods were selected to activate oocytes reconstructed during nuclear transfer with fibroblasts from mucopolysaccharidosis I- and alpha-mannosidosis-affected cats.ResultsAll treatments were able to elicit a [Ca2+]i elevation in the ooplasm with various characteristics. Pronuclear formation and development up to the blastocyst stage was most efficiently triggered by electroporation (60.5 +/- 2.9 and 11.5 +/- 1.7%) and the combined thimerosal/DTT treatment (67.7 +/- 1.8 and 10.6 +/- 1.9%); incubation of the stimulated oocytes with cycloheximide and cytochalasin B had a positive effect on embryo development. When these two methods were used to activate oocytes reconstructed during nuclear transfer, up to 84.9% of the reconstructed oocytes cleaved. When the 2 to 4-cell embryos (a total of 220) were transferred into 19 recipient females, 4 animals became pregnant. All of the fetuses developed from oocytes activated by electroporation followed by cycloheximide and cytochalasin B incubation; no fetal development was detected as a result of thimerosal/DTT activation. Although heartbeats were detected in two of the cloned fetuses, no term development occurred.ConclusionElectroporation proved to be the most effective method for the activation of cat oocytes reconstructed by nuclear transfer. The combined thimerosal/DTT treatment followed by cycloheximide and cytochalasin B incubation triggered development effectively to the blastocyst stage; whether it is a viable option to stimulate term development of cloned cat embryos needs further investigations.

show abstract

“…For the oocyte activation protocol, 5 µM of ionomycin for 5 min was used. Then, oocytes were washed one more time and cultured in 2 mM 6-dimethylaminopurine for 4 h (Méo et al, 2007). Finally, parthenogenetically activated oocytes were cultured in modified synthetic oviduct fluid (SOF) supplemented with 5% FBS, 100 IU/ml of penicillin and 50 μg/ml of streptomycin under 5% CO 2 , 5% O 2 and, 90% N 2 at 38.5°C, up to Days 7, 9 or 12, when the conditioned medium (CM) was stored at -80°C for subsequent usage.…”

Section: Oocyte Retrieval and Parthenogenetic Activationmentioning

confidence: 99%

Parthenogenetic bovine embryos secrete type I interferon capable of stimulating ISG15 in luteal cell culture

et al. 2018

View full text Add to dashboard Cite

Interferon tau (IFNT) is the pregnancy recognition signal in ruminants and is secreted by trophoblast cells. Paracrine action in the endometrium is well established by inhibiting luteolytic pulses of prostaglandin F2 alpha. Recently, endocrine action was documented in the corpus luteum, blood cell and liver. It was hypothesized that conditioned medium (CM) obtained from days 7, 9 and 12 parthenogenetic embryos alters luteal cell gene expression. The aim was to establish a bovine mixed luteal cell culture to evaluate cellular response associated to interferon stimulated genes, steroidogenesis and apoptosis. Conditioned medium was obtained from Days 7, 9 and 12 parthenogenetic (PA) embryos culture. Moreover, antiviral assay was performed on CM from Days 7, 9 and 12 to verify Type I interferon activity. Luteal cell culture was validated by steroidogenic and apoptotic genes (CYP11A1, HSD3B1, BAX, BCL2, AKT and XIAP mRNA expression), and concentration of progesterone as endpoint. Luteal cell culture was treated with interferon alpha (IFNA) and CM from parthenogenetic embryos. Antiviral assay revealed Type I interferon activity on CM from embryos increasing on Days 9 and 12. ISG15 mRNA was greater in the mixed luteal cells culture treated with 1, 10 and 100ng/ml of interferon alpha (IFNA) and also on Days 7, 9 and 12 CM treatments. Concentration of progesterone was not altered in luteal cell culture regardless of treatments. Steroidogenic and apoptotic genes were similar among groups in luteal cell culture treated with different doses of IFNA or CM from PA embryos. In conclusion, parthenogenetic embryo-derived CM has antiviral activity, luteal cell culture respond to Type I interferon by expressing IGS15. These data indicate this model can be used for IFNT endocrine signaling studies.

show abstract

Parthenogenetic activation of bovine oocytes using single and combined strontium, ionomycin and 6-dimethylaminopurine treatments

Cited by 25 publications

References 40 publications

Effect of follicular fluid supplementation during in vitro maturation on total cell number in bovine blastocysts produced in vitro

Effect of follicular fluid supplementation during in vitro maturation on total cell number in bovine blastocysts produced in vitro

Analysis of cat oocyte activation methods for the generation of feline disease models by nuclear transfer

Parthenogenetic bovine embryos secrete type I interferon capable of stimulating ISG15 in luteal cell culture

Contact Info

Product

Resources

About