In pigs, in vitro production is difficult with a high occurrence of polyspermy and low blastocyst formation rates. To test the hypothesis that this may, at least in part, be due to chromosomal errors, we employed whole genome amplification and comparative genomic hybridization, performing comprehensive chromosome analysis to assess both cells of the two-cell stage in vitro porcine embryos. We thus described the incidence, nature and origin of chromosome abnormalities, i.e. whether they derived from incorrect meiotic division during gametogenesis or aberrant mitotic division in the zygote. We observed that 19 out of 51 (37%) of two-cell stage early pig IVP embryos had a chromosome abnormality, mostly originating from an abnormal division in the zygote. Moreover, we frequently encountered multiple aneuploidies and segmental chromosome aberrations. These results indicate that the pig may be particularly sensitive to in vitro production, which may, in turn, be due to incorrect chromosome segregations during meiosis and early cleavage divisions. We thus accept our hypothesis that chromosome abnormality could explain poor IVP outcomes in pigs.
- MeSH
- aneuploidie MeSH
- chromozomální aberace * MeSH
- embryo savčí MeSH
- fertilizace in vitro veterinární MeSH
- srovnávací genomová hybridizace MeSH
- Sus scrofa embryologie genetika MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The aim of this work was to evaluate ontogeny of reactive nitrogen species (RNS) production by peripheral blood phagocytes in pig. Pig fetuses (55 and 92 days of gestation) and postnatal piglets (1, 3, 8, 17, 31 and 41 days after birth) were used. RNS production was measured by fluorescent probes diaminofluorescein-diacetate (DAF-FMDA) and dichloro-fluorescein-diacetate (H2DCFDA). Levels of nitration of cell proteins were established by immunofluorescent detection of nitrotyrosine. Levels of plasma nitrites/nitrates were detected spectrophotometrically by Griess reaction. Nitric oxide production measured by DAF-FMDA in neutrophils decreased during postnatal life. Spontaneous RNS measured by H2DCFDA decreased from 55th day of gestation to the 41st day of life. Phorbol-12-myristate-13-acetate activated production decreased during postnatal life. Production of NO measured by DAF-FMDA in macrophages decreased from the first to 41st day after birth. RNS production measured by H2DCFDA in monocytes did not show any significant changes during ontogeny. The level of nitrotyrosine significantly decreased from the third to 17th day. Levels of plasma nitrites/nitrates gradually decreased from the 55th day of gestation to the 41st day after birth. A temporary increase in all parameters occurred after weaning, but without any significance. In conclusion, RNS production has a decreasing trend during ontogeny and is transiently upregulated after weaning.
