Activation of pig oocytes using calcium ionophore: effect of protein synthesis inhibitor cycloheximide

Jílek, F.; Hűttelová, R; Petr, Jaroslav; Holubová, Monika; Rozinek, J.

doi:10.1016/s0378-4320(00)00150-0

Cited by 26 publications

(20 citation statements)

References 32 publications

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“…In this respect, the NO-dependent oocyte activation differs from activating protocols based on the elevation of intracellular levels of free calcium ions, which is successful even after a short-term treatment (several minutes) with calcium-enhancing drugs (e.g. Machatý et al, 1997;Jílek et al, 2000Jílek et al, , 2001. On the other hand, several hours of treatment are needed when the activation of pig oocytes is induced using cyclopiazonic acid, which elevates the intracellular calcium levels through the inhibition of calcium-dependent ATPases acting as intracellular sarco(endo)plasmic Ca 2+ .…”

Section: Does Nitric Oxide Activate Mammalian Oocytes?mentioning

confidence: 99%

The role of nitric oxide in parthenogenetic activation of pig oocytes: A review

Petr¹,

Eva²,

Tůmová³

et al. 2007

CZECH J ANIM SCI

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Parthenogenetic activation of mammalian oocytes with artificial stimuli is commonly applied in various reproductive biotechniques, e.g. cloning using nuclear transfer. For this reason, many studies focus on oocyte activation in vitro. Recently we have described the activation of pig oocytes using nitric oxide. This activating stimulus is very specific in many aspects. However, it does not provide an adequate stimulus for parthenogenetic development. It was shown that nitric oxide stimulated some signalling pathways which are inactive in conventional treatments for parthenogenetic activation, e.g. the cGMP-dependent signalling cascade. On the other hand, nitric oxide does not stimulate certain signalling pathways involved in oocyte activation after calcium ionophore, e.g. the PKC signalling cascade. The aim of this review is to characterize the complex processes induced in oocytes after treatment with nitric oxide. Perspectives for further research and the application of nitric oxide for parthenogenetic activation of oocytes are outlined.

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Section: Does Nitric Oxide Activate Mammalian Oocytes?mentioning

confidence: 99%

The role of nitric oxide in parthenogenetic activation of pig oocytes: A review

Petr¹,

Eva²,

Tůmová³

et al. 2007

CZECH J ANIM SCI

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“…Generally, the combination of activation stimuli (ethanol, calcium ionophore, electrical pulses, ultrasonic waves) with cycloheximide or 6-DMAP increases the activation rate of oocytes and also increases the portion of oocytes developing to the blastocyst stage (Pressice and Yang, 1994;Nussbaum and Prather, 1995;Petr et al, 1996;Jílek et al, 2000Jílek et al, , 2001Mori et al, 2008). It is not clear why under our experimental design the effect of both inhibi-tors increased the activation rate of oocytes but did not increase the proportion of oocytes developing to the blastocyst stage.…”

Section: Discussionmentioning

confidence: 81%

“…treatment using the protein synthesis inhibitor cycloheximide (Petr et al, 1996;Jílek et al, 2000;Mori et al, 2008) or treatment using the inhibitor of protein kinases 6-dimethyl aminopurine (Nussbaum and Prather, 1995;Jílek et al, 2001). The objective of this study was to enhance the efficiency of pig oocyte activation using pulsatile treatment with NO-donor combined with the inhibition of protein synthesis or inhibition of protein kinases.…”

mentioning

confidence: 99%

Effects of cycloheximide or 6-dimethyl aminopurine on the parthenogenetic activation of pig oocytes using pulsatile treatment with nitric oxide donor

Krejcova¹,

Petr²,

Krejčová³

et al. 2009

CZECH J ANIM SCI

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Pig oocytes matured in vitro were parthenogenetically activated using nitric oxide donor SNAP (2mM). Continuous treatment successfully activated the oocytes only after more than 12 hours of exposure. Pulsatile treatments during which oocytes were repeatedly exposed to 2mM SNAP for a short time (10, 20 or 30 minutes) were more efficient with regard to the activation rate, even when the total exposure time did not exceed 4 hours. Parthenogenetic development was very limited after continuous treatment with 2mM SNAP. A significantly higher proportion of developing parthenogenetic embryos was observed after the pulsatile treatment (development to the morula stage 0 vs. 18%; development to the blastocyst 0 vs. 7%; P < 0.05). However, this developmental rate was significantly lower (P < 0.05) than the development induced by conventional activation treatment with calcium ionophore (development to the morula stage, 23%; development to the blastocyst stage, 18%). When we combined pulsatile SNAP-treatment with the effect of protein kinase inhibitor 6-dimethyl aminopurine (6-DMAP) (2mM 6-DMAP for 2 hours) or with the inhibitor of protein synthesis cycloheximide (CHX) (10 µM CHX for 2 hours), we observed a significant increase (P < 0.05) in the activation rate when compared to the respective pulsatile SNAP-treatment without 6-DMAP or CHX (63 vs. 78% of activated oocytes for 6-DMAP; 63 vs. 83% of activated oocytes for CHX). However, the development of parthenogenetic embryos was not enhanced when the pulsatile SNAP-treatment was combined with 6-DMAP or with CHX.

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“…Parallel to these statements, in the present study the morulablastocyst rates in MII stage oocytes were higher than the same clusters of immature oocytes for all activation groups. Some researchers [12] found positive effects of the protein synthesis inhibitor CHX on the activation and subsequent parthenogenetic development of in vitro matured pig oocyte, activated by calcium ionophore. In our study, the protein synthesis inhibitors 6-DMAP and CHX although shown to have similar positive effects on activation rates and in terms of morula-blastocyst formation rates, 6-DMAP was more effective than CHX in both MII and imature stage oocytes.…”

Section: Discussionmentioning

confidence: 99%

“…For the attainment of low MAPK activity, electrical activation should be followed by protein synthesis inhibitors [11] . Several different activating stimulants such as ionomycin, ethanol or electrical pulses, protein synthesis inhibitors such as cycloheximide (CHX) and 6-(Dimethylamino) purine (6-DMAP) are widely used to induce the artificial activation of mammalian eggs [12] . CHX and 6-DMAP are the protein synthesis inhibitors responsible for decreasing MPF and MAPK activity of the oocyte and restarting meiosis [13] .…”

Section: Introductionmentioning

confidence: 99%

Farklı Aktivasyon Tekniklerinin Olgun Olmayan ve In Vitro Olgunlaştırılmış Kedi Oositleri Üzerine Etkisi

Demır¹,

Can²,

Ertürk³

et al. 2014

Kafkas Univ Vet Fak Derg

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SummaryThis study was conducted to determine the most successful techniques on inmature and in vitro-matured cat oocytes that were parhtenogenically activated using 6-dimethylaminopurine (6-DMAP) and cycloheximide (CHX), in combination with electrical stimulation and calcium ionophore. After 44 h of in vitro maturation, the oocytes with a polar body were separated as mature (M II) and those without a polar body were considered as immature. Four different activation treatments and two control groups were used for parthenogenetic activation with both mature and immature cat oocytes. After 48 h of activation, the oocytes were examined and the non-cleaved oocytes removed. The cleaved oocytes/embryos were cultured in vitro in mSOF medium for an additional four days. After six days of in vitro culture (IVC), embryo quality was evaluated. The results in the present study suggested that (I) both in vitro matured and immature cat oocytes have a potential to develop to morula and blastocyst stages after parthenogenetic activation, (II) electrical stimulation + 6-DMAP is a more useful technique for both matured and immature cat oocytes and (III) to our knowledge, this is the first report that describes morula and blastocyst formation from parthenogenetically activated immature cat oocytes.

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Activation of pig oocytes using calcium ionophore: effect of protein synthesis inhibitor cycloheximide

Cited by 26 publications

References 32 publications

The role of nitric oxide in parthenogenetic activation of pig oocytes: A review

The role of nitric oxide in parthenogenetic activation of pig oocytes: A review

Effects of cycloheximide or 6-dimethyl aminopurine on the parthenogenetic activation of pig oocytes using pulsatile treatment with nitric oxide donor

Farklı Aktivasyon Tekniklerinin Olgun Olmayan ve In Vitro Olgunlaştırılmış Kedi Oositleri Üzerine Etkisi

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