In vitro aging of porcine oocytes

Petrova, I. A.; Sedmı́ková, Markéta; Eva, Carola; Švestková, Dana; Rajmon, Radko

doi:10.17221/4285-cjas

Cited by 13 publications

(12 citation statements)

References 26 publications

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“…Subsequently, the oocytes were cultured in a medium supplemented with rottlerin for another 23.5 hours. For activation by nitric oxide, in vitro matured oocytes were cultured in a medium supplemented with the respective concentration of rottlerin for 30 minutes and then they were exposed to a medium supplemented with the nitric oxide donor SNAP (2mM) observed only rarely during the 24-hour ageing of porcine oocytes (Petrová et al, 2004(Petrová et al, , 2005. The involvement of individual PKC isotypes in the activation of oocytes has not been studied in greater detail.…”

Section: Discussionmentioning

confidence: 99%

Untitled

2022

Czech J. Anim. Sci.

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Porcine oocytes matured in vitro were activated for parthenogenetic development using either calcium ionophore (50µM for 10 min) or nitric oxide donor SNAP (2mM for 23.5 hours). Protein kinase C (PKC) inhibitors, bisindolylmaleimide I or rottlerin, are able to inhibit parthenogenetic activation induced by calcium ionophore. The rate of activated oocytes decreased from 69% to 2% (P < 0.05) under the effect of bisindolylmaleimide I at a concentration of 0 or 20nM, respectively. The activation rate decreased from 68% to 0% (P < 0.05) under the influence of 0 or 20µM rottlerin, respectively. PKC inhibitors Go6976 or hispidin had no effect on the oocyte activation using calcium ionophore or on oocytes activated by a nitric oxide donor. The activation of oocytes by a nitric oxide donor is not significantly influenced even under the effects of bisindolylmaleimide I or rottlerin. Based on these data we can conclude that the oocyte activation induced by calcium ionophore depends on PKC, especially on PKC-δ. On the other hand, the oocyte activation induced by nitric oxide is independent of the tested isotypes of PKC.

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Section: Discussionmentioning

confidence: 99%

Untitled

2022

Czech J. Anim. Sci.

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“…Many processes occur within an ageing oocyte that complicate or exclude the use of these oocytes, for example, they undergo spontaneous parthenogenetic activation, fragmentation or lysis (Petrová et al, 2004). Fragmentation is a manifestation of apoptosis (Tarin et al, 2001), which is an integral part of oocyte ageing (Yuce and Sadler, 2001).…”

Section: Introductionmentioning

confidence: 99%

Activation of protein kinase C suppresses fragmentation of pig oocytes aged in vitro

Petr

Krejčová

Rajmon

et al. 2011

Animal

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When cultured for an extended time, pig oocytes that matured in vitro to the stage of metaphase II undergo the complex process designated as ageing. Under our conditions, some pig oocytes aged 3 days remained at the stage of metaphase II (22%), but others underwent spontaneous parthenogenetic activation (45%), and still others perished through fragmentation (28%) or lysis (5%). Activation of protein kinases C (PKCs) using phorbol-12-myristate-13-acetate (PMA) protects oocytes from fragmentation. None of the oocytes were fragmented after 3 days of aging in 50 nM of PMA. A similar effect (8% of fragmented oocytes) was observed after a 3-day treatment of aging oocytes with 100 mM of 1-stearoyl-2arachidonoyl-sn-glycerol (STEAR). PMA and STEAR activate both calcium-dependent and calcium-independent PKCs. This combined effect on PKCs seems to be essential for the protection of oocytes from fragmentation. Neither the specific activator of calcium-dependent PKCs 1-oleoyl-2-acetyl-sn-glycerol (OLE) nor the specific activator of calcium-independent PKCs dipalmitoyl-L-a-phosphatidylinositol-3,4,5-triphosphate heptaammonium salt (DIPALM) suppressed the fragmentation of aging pig oocytes. Twenty-one percentage of oocytes fragmented when aged for 3 days in 10 mM OLE and 26% of aged oocytes fragmented in 100 nM of DIPALM. However, fragmentation was significantly suppressed to 7% when the oocytes were exposed to the combination of both 10 mM OLE and 100 nM DIPALM. Aging pig oocytes cultured for 1 day with PMA maintained a high capability of being parthenogenetically activated (86% of activated oocytes), using calcium ionophore with 6-dimethylaminopurine. Ageing oocytes treated with PMA also had high capability of cleavage (82%) after their artificial parthenogenetic activation. However, their ability to develop to the stage of blastocyst (12%) was suppressed when compared with oocytes activated immediately after their maturation (29%).

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“…(Kikuchi et al, 2000;Miao et al, 2009;Mammucari and Rizzuto, 2010 and others). Only a small percentage of oocytes remain in metaphase II arrest, while the majority of them gradually undergo parthenogenetic activation, apoptosis or lysis (Petrová et al, 2004). Spontaneous parthenogenetic activation and release from metaphase II is related to MPF inactivation, cyclin B degradation, and a decrease in MAPK activity (Sun and Nagai, 2003;Takakura et al, 2005).…”

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Inhibition of c-Jun N-terminal kinase (JNK) suppresses porcine oocyte ageing in vitro

Sedmı́ková¹,

Petr²,

Dörflerová³

et al. 2013

CZECH J ANIM SCI

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ABSTRACT:Oocyte ageing is a complex of processes that occur when matured in vitro oocytes are, after reaching the metaphase II stage, exposed to further in vitro culture. Aged oocytes remaining at the metaphase II stage undergo spontaneous parthenogenetic activation, or cellular death, through apoptosis (fragmentation) or lysis. The key factor in apoptotic pathway regulation is c-Jun-N-terminal kinase (JNK), stress kinase from the mitogene-activated protein kinase (MAPK) family. To investigate the effect of JNK inhibition on porcine oocytes ageing, cleavage rate, and embryonic development after parthenogenetic activation, DNA fragmentation, and pro-apoptotic factor Bax expression, we cultured in vitro matured oocytes for another 1-4 days in the presence of a JNK inhibitor. The inhibition of JNK significantly protected the oocytes from fragmentation (0% of fragmented oocytes under JNK inhibition vs. 13.4% of fragmented oocytes in the control group, 2 nd day of ageing) and increased the percentage of parthenogenetically activated oocytes (82 vs 57.7%, 2 nd day of ageing). The embryonic development of oocytes parthenogenetically activated after 24 h of ageing was influenced by JNK inhibition as well. The percentage of oocytes at the morula stage, after seven days of cultivation, was significantly increased when oocytes aged in the presence of a JNK inhibitor (42.5%) by comparison to the percentage of oocytes exposed to ageing in an inhibitor-free medium (23.3%). DNA fragmentation was significantly suppressed by JNK inhibition from the 1 st day of ageing, but the expression of pro-apoptotic factor Bax in the oocytes was not influenced. On the basis of our experiments, we can conclude that JNK inhibition suppresses apoptosis and DNA fragmentation of aged oocytes and improves their embryonic development following the parthenogenetic activation. However, to completely eliminate all ageing related processes is insufficient.

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In vitro aging of porcine oocytes

Cited by 13 publications

References 26 publications

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Activation of protein kinase C suppresses fragmentation of pig oocytes aged in vitro

Inhibition of c-Jun N-terminal kinase (JNK) suppresses porcine oocyte ageing in vitro

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