EGF increases expression and activity of PAs in preimplantation rat embryos and their implantation rate.
BACKGROUND
Embryo implantation plays a major role in embryogenesis and the outcome of pregnancy. Plasminogen activators (PAs) have been implicated in mammalian fertilization, early stages of development and embryo implantation. As in-vitro developing embryos resulted in lower implantation rate than those developed in-vivo we assume that a reduced PAs activity may be involved. In the present work we studied the effect of EGF on PAs activity, quantity and embryo implantation.


METHODS
Zygotes were flushed from rat oviducts on day one of pregnancy and grown in-vitro in R1ECM supplemented with EGF (10 ng/ml) and were grown up to the blastocyst stage. The control groups were grown in the same medium without EGF. The distribution and quantity of the PAs were examined using fluorescence immunohistochemistry followed by measurement of PAs activity using the chromogenic assay. Implantation rate was studied using the embryo donation model.


RESULTS
PAs distribution in the embryos was the same in EGF treated and untreated embryos. Both PAs were localized in the blastocysts' trophectoderm, supporting the assumption that PAs play a role in the implantation process in rats.EGF increased the quantity of uPA at all stages studied but the 8-cell stage as compared with controls. The tissue type PA (tPA) content was unaffected except the 8-cell stage, which was increased. The activity of uPA increased gradually towards the blastocyst stage and more so due to the presence of EGF. The activity of tPA did not vary with the advancing developmental stages although it was also increased by EGF. The presence of EGF during the preimplantation development doubled the rate of implantation of the treated group as compared with controls.
Aflalo
Eliahu D
ED
Department of Life Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel. aflaloe@bgu.ac.il
Sod-Moriah
Uriel A
UA
Potashnik
Gad
G
Har-Vardi
Iris
I
D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology; D046149:Embryo Culture Techniques; D010064:Embryo Implantation / Q000187:drug effects* / Q000502:physiology; D004624:Embryo Transfer; D047108:Embryonic Development / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005260:Female; D008297:Male; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D010959:Tissue Plasminogen Activator / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D003470:Culture Media; D004815:Epidermal Growth Factor; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator