Abstract
BackgroundThe domestic cat serves as an animal model for assisted reproductive studies of endangered felid species. To date, there are no data on the protein alterations following cryopreservation of oocytes in felid family.MethodsImmature (germinal vesicle) domestic cat oocytes were vitrified in the vitrification solution containing 35% ethylene glycol, 20% DMSO and 0.5 mM sucrose. The vitrified-warmed oocytes were matured (metaphase II) in vitro and subjected to proteomic analysis using 1DE SDS-PAGE prefractionation combined with LC–MS/MS.ResultsA total of 1712 proteins were identified in in vitro matured oocytes. Of the 1712 proteins, 1454 proteins were found in both groups, whereas, 258 proteins were differentially expressed between control and vitrified-warmed groups. In vitrified-warmed oocytes, the missing proteins were membrane and nuclear proteins; whereas, apoptosis and DNA repair proteins were overrepresented.ConclusionsThe identified missing and overexpressed proteins in vitrified-warmed oocytes represent potential markers of cryoinjuries and the developmental pathways of oocytes. The findings of differential expressed proteins may contribute to effective ways of proteome analysis of oocyte/embryo quality in order to assess safety of cryopreservation in felid species.
Highlights
The domestic cat serves as an animal model for assisted reproductive studies of endangered felid species
Immature cat oocytes were successfully cryopreserved either by vitrification or slow freezing, to improve rate of maturation and further fertilization after cryopreservation of immature oocytes, we need to study the effect of cryoinjuries, understand molecular mechanisms and proteins that involved in meiotic maturation
The present study demonstrates the effect of cryopreservation on the alterations of protein expression in domestic cat oocytes using proteomic analysis
Summary
The domestic cat serves as an animal model for assisted reproductive studies of endangered felid species. There are no data on the protein alterations following cryopreservation of oocytes in felid family. The domestic cat serves as an animal model for reproductive studies of endangered or non domestic species. Oocyte cryopreservation in mammalian species was first reported in mice [4], and subsequently in cattle [5, 6], goats [7] and humans [8]. Previous studies showed that oocytes submitted for vitrification or slow freezing induced subcellular alterations or disturbances in the intrinsic cellular homeostasis as compared to untreated controls. Vitrification in human oocytes resulted in decreased overall mRNA abundance [13]. Oocytes exposed to high concentrations of cryoprotective agents during vitrification such as ethylene glycol (EG), dimethyl sulfoxide (DMSO) or propylene glycol (PG) might
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