title: The influence of recombinant feline oviductin on different aspects of domestic cat (Felis catus) IVF and embryo quality.
abstract: Oviductin is known to be a key player providing a convenient environment for the process of fertilization affecting this by direct interaction with oocytes and sperm. As in vitro embryo production in the context of assisted reproduction for endangered felids is still in the process of optimization, oviductin might be used to improve IVF results. Recombinant His-tagged feline oviductin was expressed by transformed Escherichia coli BL21DE3 cells. The protein was purified by immobilized metal ion affinity chromatography. The effect of the recombinant protein was characterized in three experiments: a hemizona assay for sperm binding analysis, the IVF outcome, and the relative mRNA expression levels in blastocysts after IVF. A significant higher number of bound sperm cells were found after incubation in oviductin. No significant effect on cleavage, morula, and blastocyst rates with or without oviductin incubation during IVF could be observed. However, the relative mRNA abundance of GJA1, a gene, whose expression level is known to be a marker of embryo quality, was significantly increased (P value less than 0.05) in blastocysts after oviductin treatment. In contrast to this, expression of OCT4, HSP70, DNMT1, DNMT3A, BAX, IGF1R, and GAPDH was not significantly affected. We assume that our recombinant oviductin in its current nonglycosylated form is able to enhance sperm binding. Despite of a missing significant effect on IVF outcome, embryo quality in terms of relative GJA1 expression is influenced positively. These promising results demonstrate the value of recombinant oviductin for the IVF in cats.
authors.affiliation: Department Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany. Electronic address: hribal@izw-berlin.de.
authors.forename: Romy
authors.initials: R
authors.lastname: Hribal
authors.affiliation: Department Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Hachen
authors.affiliation: Department Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
authors.forename: Katarina
authors.initials: K
authors.lastname: Jewgenow
authors.affiliation: Department Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Zahmel
authors.affiliation: Department Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
authors.forename: Lorena
authors.initials: L
authors.lastname: Fernandez-Gonzalez
authors.affiliation: Department Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
authors.forename: Beate C
authors.initials: BC
authors.lastname: Braun
mesh_terms: D000818:Animals; D002415:Cats / Q000196:embryology* / Q000502:physiology; D015925:Cryopreservation / Q000662:veterinary; D046149:Embryo Culture Techniques / Q000662:veterinary*; D005307:Fertilization in Vitro / Q000662:veterinary*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology*; D012662:Semen Preservation / Q000662:veterinary; D012697:Serine Endopeptidases / Q000494:pharmacology*; D013084:Sperm-Ovum Interactions / Q000187:drug effects
chemical_list: D012333:RNA, Messenger; D011994:Recombinant Proteins; D012697:Serine Endopeptidases; C074599:oviductin
keywords: Domestic cat; Embryo quality; IVF; Oviductin; Sperm binding