Hostname: page-component-78c5997874-t5tsf Total loading time: 0 Render date: 2024-11-10T09:04:16.621Z Has data issue: false hasContentIssue false
  • English
  • Français

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: chmelikova@af.czu.cz
Get access

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 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abbott, AL and Ducibella, T 2001. Calcium and the control of mammalian cortical granule exocytosis. Frontiers in Bioscience 6, D792–D806.CrossRefGoogle ScholarPubMed
Bleil, JD, Beall, CF and Wassarman, PM 1981. Mammalian sperm-egg interaction – fertilization of mouse eggs triggers modification of the major zona pellucida glycoprotein, ZP2. Developmental Biology 86, 189–197.Google Scholar
Chantler, PD 1985. Calcium-dependent association of a protein complex with the lymphocyte plasma-membrane – probable identity with calmodulin calcineurin. Journal of Cell Biology 101, 207–216.Google Scholar
Depreux, FFS, Scheffler, JM, Grant, AL, Bidwell, CA and Gerrard, DE 2010. Molecular cloning and characterization of porcine calcineurin-alpha subunit expression in skeletal muscle. Journal of Animal Science 88, 562–571.CrossRefGoogle ScholarPubMed
Ducibella, T 1996. The cortical reaction and development of activation competence in mammalian oocytes. Human Reproduction Update 2, 29–42.Google Scholar
Dupont, G 1998. Link between fertilization-induced Ca2+oscillations and relief from metaphase II arrest in mammalian eggs: a model based on calmodulin-dependent kinase II activation. Biophysical Chemistry 72, 153–167.Google Scholar
Feine, O, Hukasova, E, Bruinsma, W, Freire, R, Fainsod, A, Gannon, J, Mahbubani, HM, Lindqvist, A and Brandeis, M 2014. Phosphorylation-mediated stabilization of Bora in mitosis coordinates Plx1/Plk1 and Cdk1 oscillations. Cell Cycle 13, 1727–1736.CrossRefGoogle ScholarPubMed
Horner, VL and Wolfner, MF 2008. Transitioning from egg to embryo: triggers and mechanisms of egg activation. Developmental Dynamics 237, 527–544.Google Scholar
Jilek, F, Huttelova, R, Petr, J, Holubova, M and Rozinek, J 2001. Activation of pig oocytes using calcium ionophore: effect of the protein kinase inhibitor 6-dimethyl aminopurine. Reproduction in Domestic Animals 36, 139–145.Google ScholarPubMed
Kheilová, K, Petr, J, Žalmanová, T, Kučerová-Chrpová, V and Řehák, D 2015. Src family kinases are involved in the meiotic maturation of porcine oocytes. Reproduction, Fertility and Development 27, 1097–1105.Google Scholar
Klee, CB, Draetta, GF and Hubbard, MJ 1988. Calcineurin. Advances in Enzymology and Related Areas of Molecular Biology 61, 149–200.Google Scholar
Krauchunas, AR, Sackton, KL and Wolfner, MF 2013. Phospho-regulation pathways during egg activation in Drosophila melanogaster . Genetics 195, 171–180+.CrossRefGoogle ScholarPubMed
Kubelka, M, Motlik, J, Schultz, RM and Pavlok, A 2000. Butyrolactone I reversibly inhibits meiotic maturation of bovine oocytes, without influencing chromosome condensation activity. Biology of Reproduction 62, 292–302.Google Scholar
Lee, J, Miyano, T and Moor, RM 2000. Localisation of phosphorylated MAP kinase during the transition from meiosis I to meiosis II in pig oocytes. Zygote 8, 119–125.CrossRefGoogle Scholar
Liu, J, Farmer, JD, Lane, WS, Friedman, J, Weissman, I and Schreiber, SL 1991. Calcineurin is a common target of cyclophilin-cyclosporine-A and FKBP-FK506 complexes. Cell 66, 807–815.Google Scholar
Longo, FJ and Chen, DY 1985. Development of cortical polarity in mouse eggs – involvement of the meiotic apparatus. Developmental Biology 107, 382–394.CrossRefGoogle ScholarPubMed
Lorca, T, Cruzalegui, FH, Fesquet, D, Cavadore, JC, Mery, J, Means, A and Doree, M 1993. Calmodulin-dependent protein kinase-II mediates inactivation of MPF and CSF upon fertilization of Xenopus eggs. Nature 366, 270–273.Google Scholar
Manders, EMM, Verbeek, FJ and Aten, JA 1993. Measurement of co-localization of objects in dual-color confocal images. Journal of Microscopy 169, 375–382.Google Scholar
Miyano, T, Dai, YF, Lee, J, Kano, K and Moor, RM 2000. Degradation of pig cyclin B1 molecules precedes MAP kinase dephosphorylation during fertilisation of the oocytes. Zygote 8, 153–158.Google Scholar
Miyazaki, S, Yuzaki, M, Nakada, K, Shirakawa, H, Nakanishi, S, Nakade, S and Mikoshiba, K 1992. Block of Ca2+wave and Ca2+oscillation by antibody to the inositol 1,4,5-trisphosphate receptor in fertilized hamster eggs. Science 257, 251–255.CrossRefGoogle Scholar
Mochida, S and Hunt, T 2007. Calcineurin is required to release Xenopus egg extracts from meiotic M phase. Nature 449, 336–340+.Google Scholar
Moos, J, Visconti, PE, Moore, GD, Schultz, RM and Kopf, GS 1995. Potential role of mitogen-activated protein-kinase in pronuclear envelope assembly and disassembly following fertilization of mouse eggs. Biology of Reproduction 53, 692–699.Google Scholar
Nishiyama, T, Yoshizaki, N, Kishimoto, T and Ohsumi, K 2007. Transient activation of calcineurin is essential to initiate embryonic development in Xenopus laevis. Nature 449, 341–345.Google Scholar
Petr, J, Chmelikova, E, Zalmanova, T, Tumova, L, Kheilova, K, Kucerova-Chrpova, V and Jilek, F 2013. Pyrethroids cypermethrin, deltamethrin and fenvalerate have different effects on in vitro maturation of pig oocytes at different stages of growth. Animal 7, 134–142.Google Scholar
Pursel, VG and Johnson, LA 1975. Freezing of boar spermatozoa – fertilizing-capacity with concentrated semen and a new thawing procedure. Journal of Animal Science 40, 99–102.Google Scholar
Romar, R, Coy, P, Gadea, J and Rath, D 2005. Effect of oviductal and cumulus cells on zona pellucida and cortical granules of porcine oocytes fertilized in vitro with epididymal spermatozoa. Animal Reproduction Science 85, 287–300.CrossRefGoogle ScholarPubMed
Rusnak, F and Mertz, P 2000. Calcineurin: form and function. Physiological Reviews 80, 1483–1521.Google Scholar
Saavedra, MD, Mondéjar, I, Coy, P, Betancourt, M, González-Márquez, H, Jiménez-Movilla, M, Avilés, M and Romar, R 2014. Calreticulin from suboolemmal vesicles affects membrane regulation of polyspermy. Reproduction 147, 369–378.Google Scholar
Saunders, CM, Larman, MG, Parrington, J, Cox, LJ, Royse, J, Blayney, LM, Swann, K and Lai, FA 2002. PLC zeta: a sperm-specific trigger of Ca(2+) oscillations in eggs and embryo development. Development 129, 3533–3544.Google Scholar
Schatten, G 1994. The centrosome and its mode of inheritance – the reduction of the centrosome during gametogenesis and its restoration during fertilization. Developmental Biology 165, 299–335.Google Scholar
Silverman-Gavrila, LB and Charlton, MP 2009. Calcineurin and cytoskeleton in low-frequency depression. Journal of Neurochemistry 109, 716–732.Google Scholar
Takeo, S, Hawley, RS and Aigaki, T 2010. Calcineurin and its regulation by Sra/RCAN is required for completion of meiosis in Drosophila . Developmental Biology 344, 957–967.CrossRefGoogle ScholarPubMed
Takeo, S, Tsuda, M, Akahori, S, Matsuo, T and Aigaki, T 2006. The calcineurin regulator Sra plays an essential role in female meiosis in Drosophila . Current Biology 16, 1435–1440.Google Scholar
Tůmová, L, Petr, J, Žalmanová, T, Chmelíková, E, Kott, T, Tichovská, H, Kučerová-Chrpová, V, Hošková, K and Jílek, F 2013. Calcineurin expression and localisation during porcine oocyte growth and meiotic maturation. Animal Reproduction Science 141, 154–163.CrossRefGoogle ScholarPubMed
Yamamuro, T, Kano, K and Naito, K 2008. Functions of FZR1 and CDC20, activators of the anaphase-promoting complex during meiotic maturation of swine oocytes. Biology of Reproduction 79, 1202–1209.CrossRefGoogle ScholarPubMed
Supplementary material: File

Tůmová supplementary material

Table S1 and Figures S1-S4

Download Tůmová supplementary material(File)
File 3.1 MB