Analysis of actinomycin D treated cattle oocytes and their use for somatic cell nuclear transfer

Moura, Marcelo Tigre; Sousa, R. V.; Leme, L. O.; Rumpf, R.

doi:10.1016/j.anireprosci.2007.10.013

Cited by 15 publications

(18 citation statements)

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“…To induce apoptosis, we used actinomycin D which is known as an inducer of apoptosis on different cell types by inhibiting DNA transcription ( 11 , 27 ). Some articles have shown that actinomycin D decreases development and cleavage rate of embryos, and increases apoptotic cells in blastocyst ( 10 , 11 , 28 ).…”

Section: Discussionmentioning

confidence: 99%

Effect of quercetin on the number of blastomeres, zona pellucida thickness, and hatching rate of mouse embryos exposed to actinomycin D: An experimental study

Sameni

Javadinia

Safari

et al. 2018

IJRM

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Background: Quercetin is a flavonoid with the ability to improve the growth of embryos in vitro, and actinomycin D is an inducer of apoptosis in embryonic cells. Objective: The aim was to evaluate the effect of quercetin on the number of viable and apoptotic cells, the zona pellucida (ZP) thickness and the hatching rate of preimplantation embryos exposed to actinomycin D in mice. Materials and Methods: Two-cell embryos were randomly divided into four groups (Control, Quercetin, actinomycin D, and Quercetin + actinomycin D group). Blastocysts percentage, hatched blastocysts, and ZP thickness of blastocysts was measured. The number of blastomeres was counted by Hoechst and propidium iodide staining and the apoptotic cells number was counted by TUNEL assay. Results:The results showed that the use of quercetin significantly improved the growth of embryos compared to the control group (p=0.037). Moreover, quercetin reduced the destructive effects of actinomycin D on the growth of embryos significantly (p=0.026). Conclusion: quercetin may protect the embryos against actinomycin D so that increases the number of viable cells and decreases the number of apoptotic cells, which can help the expansion of the blastocysts, thinning of the ZP thickness and increasing the hatching rate in mouse embryos.

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Section: Discussionmentioning

confidence: 99%

Effect of quercetin on the number of blastomeres, zona pellucida thickness, and hatching rate of mouse embryos exposed to actinomycin D: An experimental study

Sameni

Javadinia

Safari

et al. 2018

IJRM

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“…Cumulus-oocyte complexes (COC) were retrieved from 2-6 mm follicles with a 10 G needle into 10 mL syringes, and follicular content was deposited in Petri dishes containing HEPES-buffered IVM medium (H-IVM), consisting of TCM199 with Hank's salts, 10 % fetal bovine serum (FBS), 50 IU mL -1 heparin and 50 µg mL -1 gentamicin (In vitro Brasil). The COCs were recovered and selected those with a homogeneous cytoplasm granulation and three or more cumulus cell layers (Moura et al, 2008), and further washed in IVM medium. The IVM medium was composed of TCM-199 with Earle's salts supplemented with 10 % FBS, 2 mM Lglutamine, 5.0 μg mL -1 FSH (Sigma ® ), and 50 µg mL -1 gentamicin.…”

Section: Materials and Methods Oocyte Recovery And In Vitro Maturationmentioning

confidence: 99%

“…Parthenogenetic activation (PA) was performed as previously described by Moura et al (2008). The COCs were denuded with 0.2 % hyaluronidase and gentle pipetting.…”

Section: Parthenogenetic Activationmentioning

confidence: 99%

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Expression of RONIN and NANOG-associated proteins in goat parthenogenetic embryos

et al. 2017

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The expression of a subset of transcription factors is enriched in early preimplantation embryos, which contributes to their cellular plasticity. RONIN, NANOG and its associated proteins are PluripotencyAssociated Transcription Factors (PATF) that control relevant downstream pathways in pluripotent stem cells, but their activity in early embryos remained less understood. The work was aimed to determine the expression of RONIN and four NANOG-associated PATFs in goat preimplantation embryos. Goat embryos were produced in vitro by parthenogenetic activation. Gene transcripts of cleavage-stage embryos were investigated by reverse transcriptase-polymerase chain reaction (RT-PCR), while blastocysts were analyzed by both RT-PCR and quantitative RT-PCR (RT-qPCR) assays. Gene transcripts of ZFP281, NAC1, and NR0B1 were detected in cleavage-stage embryos, while RONIN and OCT4 were not found expressed. Detection in blastocysts by RT-PCR confirmed the activity of NR0B1, RONIN, and OCT4. Moreover, all five PATF were detected in blastocysts by RT-qPCR (ZFP281, NAC1, RONIN, OCT4, and NR0B1). In conclusion, RONIN and NANOG-associated proteins are active during goat parthenogenetic preimplantation development and hold stage-specific expression patterns.Keywords: Capra hircus; caprine; embryogenesis; RT-PCR; RT-qPCR; mRNA. ResumoA expressão de um conjunto de fatores de transcrição é aumentada em embriões antes da implantação, fato que contribui para a sua plasticidade celular. RONIN, NANOG e suas proteínas associadas são Fatores de transcrição relacionados à pluripotência (FTRP) que controlam vias de sinalização importantes para células-tronco pluripotentes, embora suas atividades em embriões permaneçam menos compreendidas. O objetivo do trabalho foi determinar a expressão de RONIN e quatro FTRPs associados ao NANOG em embriões caprinos. Os embriões foram produzidos in vitro por ativação partenogenética. Embriões na fase de clivagem foram investigados por transcrição reversa, seguida da reação da reação em cadeia da polimerase (RT-PCR), enquanto que os blastocistos foram analisados por RT-PCR e por RT-PCR quantitativa (RT-qPCR). Os transcritos de ZFP281, NAC1 e NR0B1 foram detectados em embriões clivados, enquanto que RONIN e OCT4 não foram encontrados. A detecção em blastocistos pela RT-PCR confirmou a atividade de NR0B1, RONIN e OCT4. Além disso, os cinco FTRPS foram detectados em blastocistos pela RT-qPCR (ZFP281, NAC1, RONIN, OCT4 e NR0B1). Em conclusão, RONIN e os FTRP associados ao NANOG são ativos durante o desenvolvimento pré-implantacional de caprinos e apresentam padrão de expressão estádio-específica.

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“…(Hosseini et al, 2015). Moura e colaboradores (2008), avaliaram a utilização da actinomicina D como método alternativo para a enucleação química e mostraram que apesar de induzir inativação dos cromossomos, essa substância causa uma redução drástica na taxa de blastocisto (Moura et al, 2008).…”

Section: Enucleação Reconstrução E Ativação Dos Ovócitosunclassified

Bloqueio meiótico como alternativa para aumentar a produção de embriões clone por transferência nuclear

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Analysis of actinomycin D treated cattle oocytes and their use for somatic cell nuclear transfer

Cited by 15 publications

References 37 publications

Effect of quercetin on the number of blastomeres, zona pellucida thickness, and hatching rate of mouse embryos exposed to actinomycin D: An experimental study

Effect of quercetin on the number of blastomeres, zona pellucida thickness, and hatching rate of mouse embryos exposed to actinomycin D: An experimental study

Expression of RONIN and NANOG-associated proteins in goat parthenogenetic embryos

Bloqueio meiótico como alternativa para aumentar a produção de embriões clone por transferência nuclear

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