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

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 activa...

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Main Authors: J. Petr, M. Krejčová, R. Rajmon, F. Jílek
Format: Article
Language:English
Published: Elsevier 2011-01-01
Series:Animal
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S1751731110002119
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author J. Petr
M. Krejčová
R. Rajmon
F. Jílek
author_facet J. Petr
M. Krejčová
R. Rajmon
F. Jílek
author_sort J. Petr
collection DOAJ
description 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 μM 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-α-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 μM 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 μM 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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spelling doaj.art-5fbb0f63b08240469d590dd4c175c7932022-12-21T19:23:00ZengElsevierAnimal1751-73112011-01-0154565571Activation of protein kinase C suppresses fragmentation of pig oocytes aged in vitroJ. Petr0M. Krejčová1R. Rajmon2F. Jílek3Research Institute of Animal Production, Přátelství 815, 10401 Prague 10-Uhříněves, Czech RepublicFaculty of Agrobiology, Food and Natural Resources, Department of Veterinary Sciences, Czech University of Life Sciences Prague, Kamýcká 129, 165 21 Prague 6 – Suchdol, Czech RepublicFaculty of Agrobiology, Food and Natural Resources, Department of Veterinary Sciences, Czech University of Life Sciences Prague, Kamýcká 129, 165 21 Prague 6 – Suchdol, Czech RepublicFaculty of Agrobiology, Food and Natural Resources, Department of Veterinary Sciences, Czech University of Life Sciences Prague, Kamýcká 129, 165 21 Prague 6 – Suchdol, Czech RepublicWhen 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 μM 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-α-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 μM 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 μM 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%).http://www.sciencedirect.com/science/article/pii/S1751731110002119pig oocyteoocyteageingoocyte fragmentationprotein kinase C
spellingShingle J. Petr
M. Krejčová
R. Rajmon
F. Jílek
Activation of protein kinase C suppresses fragmentation of pig oocytes aged in vitro
Animal
pig oocyte
oocyte
ageing
oocyte fragmentation
protein kinase C
title Activation of protein kinase C suppresses fragmentation of pig oocytes aged in vitro
title_full Activation of protein kinase C suppresses fragmentation of pig oocytes aged in vitro
title_fullStr Activation of protein kinase C suppresses fragmentation of pig oocytes aged in vitro
title_full_unstemmed Activation of protein kinase C suppresses fragmentation of pig oocytes aged in vitro
title_short Activation of protein kinase C suppresses fragmentation of pig oocytes aged in vitro
title_sort activation of protein kinase c suppresses fragmentation of pig oocytes aged in vitro
topic pig oocyte
oocyte
ageing
oocyte fragmentation
protein kinase C
url http://www.sciencedirect.com/science/article/pii/S1751731110002119
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AT mkrejcova activationofproteinkinasecsuppressesfragmentationofpigoocytesagedinvitro
AT rrajmon activationofproteinkinasecsuppressesfragmentationofpigoocytesagedinvitro
AT fjilek activationofproteinkinasecsuppressesfragmentationofpigoocytesagedinvitro