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The application of in vitro cattle embryo production system to study the influence of elevated temperature on oocyte maturation, fertilization and early embryonic development

100%
Rynkowska A., Rapa?a L., Trzeciak P., Duszewska A. M.
Biotechnologia
|
2011
|
issue 1
45-53
EN
Higher air temperature in summer causes a significant reduction in fertility in cattle. Increase in female body temperature during the period of reproduction by only 2EC, also known as hyperthermia, leads to disturbances in the functioning of the female reproductive system, oocytes maturation, fertilization and embryos development. Particularly sensitive to high temperatures are embryos in the first and second day after fertilization (thermosensitive), but just at third till fifth day after fertilization their resistance to thermal stress significantly increases. Morula-stage and blastocyst-stage bovine embryos are insensitive to elevated temperatures (thermoresistant). Most probably this is due to the increasing number of cells within the embryo and the capacity to activate defense mechanisms based on the synthesis of various factors providing resistance to high temperatures. These factors include heat shock protein 70 (HSP70), antioxidants such as glutathione, and IGF-1. One of the responses of the embryo to elevated temperature is the induction of apoptosis, which is associated with the activation of embryonic genome. Owing to the apoptosis, cells damaged by high temperature may be eliminated from the embryo, which increases their chance of survival. Precise examination of the mechanisms responsible for the development of thermotolerance of preimplantation bovine embryos will enable their protection from the consequences of elevated temperature. The aim of this review is to summarise experiments in which in vitro embryo production system was used to estimate the influence of elevated temperature on cattle fertility.
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The application of in vitro cattle embryo production system to study the influence of elevated temperature on oocyte maturation, fertilization and early embryonic development

100%
Rynkowska A., Rąpała Ł., Trzeciak P., Duszewska A. M.
BioTechnologia
|
2011
|
issue 1
3

Noninvasive fluorescent screening of microinjected bovine embryos to predict transgene integration

76%
Rosochacki S. J., Kozikova L., Korwin-Kossakowski M., Matejczyk M., Poloszynowicz J., Duszewska A. M.
Folia Biologica
|
2003
|
vol. 51
|
issue 1-2
97-104
EN
Most transgenic domestic animals are generated by direct microinjection of DNA fragments into zygote pronuclei. It has generally been assumed that the majority of integration events should occur prior to the first round of chromosomal DNA replication. The aim of this study was to investigate the expression of GFP in bovine preimplantation embryos by using a gfp reporter gene consisting of chicken beta-actin promoter, the CMV-IE enhancer, gfp cDNA (EGFP) (732 bp) and rabbit beta-globin polyadenylation sequences. In five experiments 302 bovine zygotes were injected while 75 served as a control. The fluorescence intensity was detected at 72 and 168 h following fertilization in bovine embryos injected with 3 ng/mu l in experiments 1-3, and injected with 5 ng/mu l in experiments 4-5. Eight embryos were considered as expressing green fluorescence protein; 2 of them were 100% fluorescent after microinjection of a higher dose of the DNA; one was 75%, two - 50%, and three 25% transgenic. The mosaicism was assumed to be at 75%. The results indicated that the fluorescence could be analyzed at any time of bovine embryo development. It was therefore concluded, that chicken -actin promoter together with the CMV-IE enhancer would confer a strong expression of the gfp reporter gene in preimplantation bovine embryos. Therefore, using GFP that could be simply detected in live bovine (transgenic) embryos would be very promising in establishing transgenic lines of domestic animals producing in their fluids human therapeutic proteins.
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