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• W2740276597 abstract "Apoptosis has an important role in pre-implantation embryo development because embryonic cells are subjected to DNA damage that can activate repair mechanisms or induce apoptosis. The latter culminates in the activation of the caspase enzyme responsible for cleaving various cellular substrates. The aim of this study was to measure differences in Casp15 gene expression between Piau embryos and embryos from a swine commercial line. Gilts were slaughtered or subjected to exploratory laparotomy for recovery of embryos on day 6 after mating (day 0 = estrus). Surgery was performed as previously described (RD Cameron 1989 Australian Veterinary Journal, 66, 314- 318) and approved by the Ethics Committee of the UFV. Commercial line embryos were obtained by flushing the uterine tract post mortem 6 days post mating, using PBS at 37 ° C plus 1% PVA. Embryos were classified according to the recommendations of the IETS (International Embryo Transfer Society, 1998) and 65 grade I blastocysts were selected for the study (35 and 30 to Piau and commercial line, respectively). The embryos were preserved in 2 mL of PBS with 1% PVA in cryotubes, and kept in liquid nitrogen until RNA extraction. The extraction was performed using RNeasy Mini Kit (Qiagem, Valencia, CA). Total RNA was reverse transcribed with M-MuLV ProtoScript First Strand cDNA Synthesis kit (Biolabs, Ipswich, MA). Real-time polymerase chain reaction was performed using GoTaq qPCR Master Mix (Promega, Madison, WI) in an ABI PRISM 7300 Sequence Detection Systems (Applied Biosystems, Foster City, CA). Primers were designed from pig sequences available in GenBank. The linearity of Casp15 mRNA amplification was similar to reference gene (GAPDH). The reactions were performed using 200 nM and 400 nM primers for Casp15 and GAPDH, respectively, and 1.6 ng of cDNA reaction for each gene. The thermocycling conditions consisted of 40 cycles of 30 sec melting at 95 °C followed by 30 seconds annealing and extending at 60 °C. After amplification, an analysis of the melting curve was performed to confirm the absence of unspecific products . Gene expression data were evaluated using the 2-ΔCt method (KJ Livak and Schmittgen TD 2001, Methods, 25, 402-408). The results of gene expression were analyzed using the GLM procedure (SAS Institute, Cary, NC). The Casp15 mRNA expression did not differ between the preimplantation embryos from Piau and commercial line (P > 0.05). Despite the fact that no difference was found, the development of Piau embryo and litter size are different compared to other races." @default.
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• W2740276597 date "2016-03-01" @default.
• W2740276597 modified "2023-09-22" @default.
• W2740276597 title "Caspase15 gene expression in swine embryos of different genetic groups" @default.
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