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Mirjan Thys Reproduction, Virology, Ghent Research Group on Nanomedicines, Departments of

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Hans Nauwynck Reproduction, Virology, Ghent Research Group on Nanomedicines, Departments of

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Dominiek Maes Reproduction, Virology, Ghent Research Group on Nanomedicines, Departments of

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Maarten Hoogewijs Reproduction, Virology, Ghent Research Group on Nanomedicines, Departments of

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Dries Vercauteren Reproduction, Virology, Ghent Research Group on Nanomedicines, Departments of

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Tom Rijsselaere Reproduction, Virology, Ghent Research Group on Nanomedicines, Departments of

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Herman Favoreel Reproduction, Virology, Ghent Research Group on Nanomedicines, Departments of

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Ann Van Soom Reproduction, Virology, Ghent Research Group on Nanomedicines, Departments of

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Fibronectin (Fn) is a 440 kDa glycoprotein assumed to participate in sperm–egg interaction in human. Recently, it has been demonstrated that Fn – when present during bovine IVF – strongly inhibits sperm penetration. The present study was conducted firstly to evaluate the expression of Fn and its integrin receptor (α5β1) on male and female bovine gametes using indirect immunofluorescence and secondly, to determine the function of Fn during bovine IVF. Endogenous Fn was detected underneath the zona pellucida (ZP) and integrin α5 on the oolemma of cumulus-denuded oocytes. Bovine spermatozoa displayed integrin α5 at their equatorial segment after acrosome reaction. We established that the main inhibitory effect of exogenously supplemented Fn was located at the sperm–oolemma binding, with a (concurrent) effect on fusion, and this can probably be attributed to the binding of Fn to spermatozoa at the equatorial segment, as shown by means of Alexa Fluor 488-conjugated Fn. Combining these results, the inhibitory effect of exogenously supplemented Fn seemed to be exerted on the male gamete by binding to the exposed integrin α5β1 receptor after acrosome reaction. The presence of endogenous Fn underneath the ZP together with integrin α5 expression on oolemma and acrosome-reacted (AR) sperm cell surface suggests a ‘velcro’ interaction between the endogenous Fn ligand and corresponding receptors on both (AR) sperm cell and oolemma, initiating sperm–egg binding.

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Katrien Smits
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Jan Govaere
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Luc J Peelman Department of Reproduction, Department of Nutrition, Laboratory of Zoophysiology, Department of Pharmaceutics, Department of Equine Sciences, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium

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Karen Goossens Department of Reproduction, Department of Nutrition, Laboratory of Zoophysiology, Department of Pharmaceutics, Department of Equine Sciences, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium

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Dirk C de Graaf Department of Reproduction, Department of Nutrition, Laboratory of Zoophysiology, Department of Pharmaceutics, Department of Equine Sciences, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium

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Dries Vercauteren Department of Reproduction, Department of Nutrition, Laboratory of Zoophysiology, Department of Pharmaceutics, Department of Equine Sciences, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium

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Leen Vandaele
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Maarten Hoogewijs
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Eline Wydooghe
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Tom Stout Department of Reproduction, Department of Nutrition, Laboratory of Zoophysiology, Department of Pharmaceutics, Department of Equine Sciences, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium

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Ann Van Soom
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The necessity for early interaction between the embryo and the oviductal and/or uterine environment in the horse is reflected by several striking differences between equine embryos that develop in vivo and those produced in vitro. Better understanding of the salient interactions may help to improve the efficiency of in vitro equine embryo production. In an initial experiment, cleavage-stage in vitro-produced (IVP) equine embryos were transferred into the uterus of recipient mares that had ovulated recently to determine whether premature placement in this in vivo environment would improve subsequent development. In a second experiment, an important element of the uterine environment was mimicked by adding uterocalin, a major component of the endometrial secretions during early pregnancy, to the culture medium. Intrauterine transfer of cleavage-stage IVP equine embryos yielded neither ultrasonographically detectable pregnancies nor day 7 blastocysts, indicating that the uterus is not a suitable environment for pre-compact morula stage horse embryos. By contrast, exposure to uterocalin during IVP improved capsule formation, although it did not measurably affect the development or expression of a panel of genes known to differ between in vivo and in vitro embryos. Further studies are required to evaluate whether uterocalin serves purely as a carrier protein or more directly promotes improved capsule development.

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