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The effect of FF-MAS on porcine cumulus–oocyte complex maturation, fertilization and pronucleus formation in vitro

Published online by Cambridge University Press:  01 August 2006

Inger Faerge ,
Frantisek Strejcek ,
Jozef Laurincik ,
Detlef Rath ,
Heiner Niemann ,
Karl Schellander ,
Christine Rosenkranz ,
Poul Maddox Hyttel  and
Christian Grøndahl
Inger Faerge*
Affiliation:
Fertility Team, Novo Nordisk A/S, Copenhagen, Denmark.
Frantisek Strejcek
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic.
Jozef Laurincik
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic. Research Institute of Animal Production, Nitra, Slovak Republic.
Detlef Rath
Affiliation:
Research Institute of Animal Production, Nitra, Slovak Republic.
Heiner Niemann
Affiliation:
Research Institute of Animal Production, Nitra, Slovak Republic.
Karl Schellander
Affiliation:
Faculty of Agriculture, Institute of Animal Breeding Science, University of Bonn, Bonn, Germany.
Christine Rosenkranz
Affiliation:
University of Veterinary Medicine, Vienna, Austria.
Poul Maddox Hyttel
Affiliation:
Veterinary and Agricultural University, Copenhagen, Denmark.
Christian Grøndahl
Affiliation:
Fertility Team, Novo Nordisk A/S, Copenhagen, Denmark.
*
All correspondence to: Inger Faerge, DVM, PhD, Institute of Basic Veterinary Sciences, Groennegaardsvej 7, 1870 Frb C, Copenhagen, Denmark. Tel: +45 35283882. Fax: +45 35282548. e-mail: [email protected]
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Summary

Follicular fluid meiosis-activating sterol (FF-MAS) has been isolated from the follicular fluid (FF) of several species including man. FF-MAS increases the quality of in vitro oocyte maturation, and thus the developmental potential of oocytes exposed to FF-MAS during in vitro maturation is improved. The aim of the present study was to investigate the effects of FF-MAS on porcine oocyte maturation and pronucleus formation in vitro. Porcine cumulus–oocyte complexes (COCs) were isolated from abattoir ovaries and in vitro matured for 48 h in NCSU 37 medium supplemented with 1 mg/l cysteine, 10 ng/ml epidermal growth factor and 50 μM 2-mercaptoethanol with or without 10% porcine follicular fluid (pFF). For the first 22 h, 1 mM db-cAMP and 10 I.E PMSG/hCG was added. The medium was supplemented with 1 μM, 3 μM, 10 μM, 30 μM or 100 μM FF-MAS dissolved in ethanol. After maturation the COCs were denuded mechanically using a fine glass pipette under constant pH and in vitro fertilized with fresh semen (5 × 105 spermatozoa/ml). The presumptive zygotes were evaluated 18 h after fertilization. The addition of pFF increased the monospermic as well as the polyspermic penetration of oocytes. In the absence of pFF, the addition of FF-MAS decreased the polyspermic penetration rate, whereas FF-MAS in combination with pFF decreased monospermic and increased polyspermic penetration. The degeneration rate of ova decreased in the presence of FF-MAS irrespective of the presence or absence of pFF. In the absence of pFF, FF-MAS at 3–10 μM increased the number of zygotes with advanced maternal pronuclear stages. In supraphysiological doses, i.e. 30–100 μM, FF-MAS dose-dependently and reversibly inhibited nuclear maturation in the absence of pFF.

Keywords

FF-MASPigPronucleus formationSterols
Type
Research Article
Information
Zygote , Volume 14 , Issue 3 , August 2006 , pp. 189 - 199
Copyright
Copyright © Cambridge University Press 2006

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