Analysis of transgenic rabbit vitrified embryos carrying EGFP gene
2011
Chrenek, P., Slovak Univ. of Agriculture in Nitra (Slovak Republic) | Makarevich, A.V., Slovak Univ. of Agriculture in Nitra (Slovak Republic)
The aim of our study was to investigate influence of vitrification on developmental rate and quality (total cells number, numer of ICM blastomeres, apoptotic index and embryo diameter) of rabbit transgenic embryos carrying exogenous EGFP gene. EGFP positive rabbit embryos were produced under in vitro conditions by the microinjection of foreign gene into the pronucleus of fertilized eggs. EGFP expression was detected by fluorescent microscope (at 492 nm). Developmental rate of vitrified/thawed transgenic embryos reached hatching blastocyst stage (68.00 %) differed significantly (p less than 0.001) from those in control embryo (100.00 % resp.). The higher proportion of ICM cells (32.00 %) and embryo diameter (130.85 plus minus 10.90) were found in the kontrol group compared to transgenic ones. The ratio of apoptotic cells was higher (2.90 %) in vitrified EGFP-positive embryos compared with the control group (2.50 %). Our results demonstrate, that neither gene microinjection itself, nor exogenous (EGFP) gene expression do interfere with quality of rabbit embryos. However, combination of microinjection and vitrification significantly decreases (p less than 0.001) survival rate of rabbit embryos.
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