[en] The objective of this study was to test the accuracy of genotype diagnosis after whole amplification of DNA extracted from biopsies obtained by trimming goat embryos and to evaluate the viability of biopsied embryos after vitrification/warming and transfer. Whole genome amplification (WGA) was performed using Multiple Displacement Amplification (MDA). Sex and prion protein (PRNP) genotypes were determined. Sex diagnosis was carried out by PCR amplification of ZFX/ZFY and Y chromosome-specific sequences. Prion protein genotype determination was performed on codons 142, 154, 211, 222 and 240. Embryos were collected at day 7 after oestrus and biopsied either immediately after collection (blastocysts and expanded blastocysts) or after 24 h of in vitro culture (compacted morulae). Biopsied embryos were frozen by vitrification. Vitrified whole embryos were kept as control. DNA of biopsies was extracted and amplified using MDA. Sex diagnosis was efficient for 97.4% of biopsies and PRNP genotyping was determined in 78.7% of biopsies. After embryo transfer, no significant difference was observed in kidding rate between biopsied and vitrified control embryos, whereas embryo survival rate was different between biopsied and whole vitrified embryos (p = 0.032). At birth, 100% of diagnosed sex and 98.2% of predetermined codons were correct. Offspring PRNP profiles were in agreement with parental genotype. Whole genome amplification with MDA kit coupled with sex diagnosis and PRNP genotype predetermination are very accurate techniques to genotype goat embryos before transfer. These novel results allow us to plan selection of scrapie-resistant genotypes and kid sex before transfer of cryopreserved embryo.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Guignot, F.; Institut Scientifique de Recherche Agronomique - INRA > UMR85 Physiologie de la Reproduction et des Comportements
Perreau, C.; Institut Scientifique de Recherche Agronomique - INRA > UMR85 Physiologie de la Reproduction et des Comportements
Cavarroc, C.; Institut Scientifique de Recherche Agronomique - INRA > UMR85 Physiologie de la Reproduction et des Comportements
Touze, J. L.; Institut Scientifique de Recherche Agronomique - INRA > UMR85 Physiologie de la Reproduction et des Comportements
Pougnard, J. L.; Institut Scientifique de Recherche Agronomique - INRA > UE88 Insémination caprine et porcine, Rouillé, France
Dupont, F.; Institut Scientifique de Recherche Agronomique - INRA > UMR85 Physiologie de la Reproduction et des Comportements
Beckers, Jean-François ; Université de Liège - ULiège > Département de sciences fonctionnelles > Physiologie de la reproduction
Remy, B.; Université de Liège - ULiège
Babilliot, J. M.; LABOGENA
Bed'hom, B.; Institut Scientifique de Recherche Agronomique - INRA > UMR1313 Génétique-Animale et Biologie Intégrative, Jouy en Josas, France
Lamoriniere, J.; Goat breeder, Neuillé Pont Pierre, France
Mermillod, P.; Institut Scientifique de Recherche Agronomique - INRA > UMR85 Physiologie de la Reproduction et des Comportements
Baril, G.; Institut Scientifique de Recherche Agronomique - INRA > UMR85, Physiologie de la Reproduction et des Comportements
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