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Calcineurin role in porcine oocyte activation

Published online by Cambridge University Press:  10 May 2016

L. Tůmová ,
E. Chmelíková ,
T. Žalmanová ,
V. Kučerová-Chrpová ,
R. Romar ,
M. Dvořáková ,
K. Hošková  and
J. Petr
L. Tůmová
Affiliation:
Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences in Prague, 165 21 Prague 6 – Suchdol, Czech Republic
E. Chmelíková*
Affiliation:
Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences in Prague, 165 21 Prague 6 – Suchdol, Czech Republic
T. Žalmanová
Affiliation:
Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences in Prague, 165 21 Prague 6 – Suchdol, Czech Republic
V. Kučerová-Chrpová
Affiliation:
Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences in Prague, 165 21 Prague 6 – Suchdol, Czech Republic
R. Romar
Affiliation:
Department of Physiology, Faculty of Veterinary, Campus Mare Nostrum, University of Murcia,IMIB-Arrixaca,30100 Murcia, Spain
M. Dvořáková
Affiliation:
Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences in Prague, 165 21 Prague 6 – Suchdol, Czech Republic
K. Hošková
Affiliation:
Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences in Prague, 165 21 Prague 6 – Suchdol, Czech Republic
J. Petr
Affiliation:
Institute of Animal Science, 104 00 Prague 22-Uhříněves, Czech Republic
*
†E-mail: [email protected]
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Abstract

Calcineurin is required for oocyte exit from meiotic block in metaphase II (MII) stage in invertebrates and also in lower vertebrates. However, the role of calcineurin in mammalian oocyte activation is still unclear. The aim of this study was to determine whether calcineurin is involved in the processes regulating porcine oocyte activation. Indirect immunofluorescence demonstrated localization of both calcineurin subunits, CnA and CnB, especially in the cortex area of MII oocytes, in vitro fertilized and also parthenogenetically activated oocytes. After activation, the fluorescence intensity of the protein in the cortex area of oocytes remains unchanged; the protein calcineurin in the cytoplasm was recorded mainly around the pronuclei. Treatment of matured oocytes with calcineurin inhibitors, cyclosporin A (CsA) and hymenistatin I (HS-I), followed by activation with calcium ionophore A23187, significantly decreased the rate of activated oocytes compared to oocytes that were treated only with calcium ionophore (Ca-Io), (CsA+Ca-Io 25.0% v. Ca-Io 83.3%; HS-I+Ca-Io 32.5% v. Ca-Io 85.0%). Compared to the control, CsA treatment of matured oocytes followed by activation with Ca-Io did not affect the activity level of metaphase-promoting factor (MPF) and mitogen-activated protein kinase (MAPK) in activated oocytes evaluated by kinase activity assay. Simultaneous staining of calcineurin and cortical granule content in matured oocytes showed that calcineurin distributed in the cortical area of the oocyte has not been colocalized with cortical granules content. On the other hand, the calcineurin inhibition before parthenogenetic activation leads to a reduction of the cortical reaction level compared to oocytes that were not treated with CsA (complete exocytosis: CsA+Ca-Io 2.6% v. Ca-Io 83.9%; sum of cortical granule brightness: CsA + Ca-Io 0.69 v. Ca-Io 0.15). Our results showed that calcineurin is involved in the process of pig oocyte activation and cortical granule exocytosis; however this regulation seems to be MPF and MAPK independent.

Keywords

pigoocyteactivationcortical granule exocytosiscalcineurin
Type
Research Article
Information
animal , Volume 10 , Issue 12 , December 2016 , pp. 1998 - 2007
Copyright
© The Animal Consortium 2016 

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