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Expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro

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. The invasion of trophoblast cells into the endometrium during the impla...

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Autores principales: Aflalo, Eliahu D, Sod-Moriah, Uriel A, Potashnik, Gad, Har-Vardi, Iris
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC549534/
https://www.ncbi.nlm.nih.gov/pubmed/15703084
http://dx.doi.org/10.1186/1477-7827-3-7
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author Aflalo, Eliahu D
Sod-Moriah, Uriel A
Potashnik, Gad
Har-Vardi, Iris
author_facet Aflalo, Eliahu D
Sod-Moriah, Uriel A
Potashnik, Gad
Har-Vardi, Iris
author_sort Aflalo, Eliahu D
collection PubMed
description 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. The invasion of trophoblast cells into the endometrium during the implantation process can be blocked by inhibitors of serine proteases, illustrating the role of these enzymes in the invasion process. As in vitro developing embryos resulted in lower implantation rate than those developed in vivo we assume that a reduced PAs activity may lead to it. There is hardly any information regarding qualitative or quantitative differences in expression of PAs in preimplantation embryos, or comparisons between in vivo and in vitro developed embryos. The purpose of this study was to assess the expression of urokinase type (uPA) and tissue type (tPA) plasminogen activators in in vivo and in vitro preimplantation development in rat embryos using immunofluorescence confocal microscopy and computerized image analysis. METHODS: Zygotes, 2-cell, 4-cell, 8-cell, morula and blastocyst stages of development were flushed from the reproductive tract (control groups) of Wistar rats. Zygotes were flushed and grown in vitro to the above mentioned developmental stages and comprised the experimental groups. Immunofluorescence microscopy and computerized image analysis were used to evaluate both qualitative (localization) and quantitative expression of plasminogen activators. RESULTS: uPA and tPA were found to be expressed in rat embryos throughout their preimplantation development, both in vivo and in vitro. While uPA was localized mainly in the cell cytoplasm, the tPA was detected mainly on cell surface and in the perivitelline space. In blastocysts, both in vivo and in vitro, uPA and tPA were localized in the trophectoderm cells. Total uPA content per embryo was higher in the in vivo as compared with the in vitro developed embryos at all stages measured. Blastocyst uPA content was significantly low as compared with the four-cell, eight-cell, and morula stages. Total tPA content was higher in embryos developed in vivo than those developed in vitro except for the 4-cell and 8-cell stages. CONCLUSION: In vitro embryo development leads to lower PAs expression in a stage dependent manner as compared with in vivo developing controls. The enzymes studied vary probably in the ratio of their active and inactive forms as there is no correlation between their content and the activity observed in our previous study. The localization of both PAs in the blastocysts' trophectoderm supports the assumption that PAs plays a role in the implantation process in rats.
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spelling pubmed-5495342005-02-25 Expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro Aflalo, Eliahu D Sod-Moriah, Uriel A Potashnik, Gad Har-Vardi, Iris Reprod Biol Endocrinol Research 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. The invasion of trophoblast cells into the endometrium during the implantation process can be blocked by inhibitors of serine proteases, illustrating the role of these enzymes in the invasion process. As in vitro developing embryos resulted in lower implantation rate than those developed in vivo we assume that a reduced PAs activity may lead to it. There is hardly any information regarding qualitative or quantitative differences in expression of PAs in preimplantation embryos, or comparisons between in vivo and in vitro developed embryos. The purpose of this study was to assess the expression of urokinase type (uPA) and tissue type (tPA) plasminogen activators in in vivo and in vitro preimplantation development in rat embryos using immunofluorescence confocal microscopy and computerized image analysis. METHODS: Zygotes, 2-cell, 4-cell, 8-cell, morula and blastocyst stages of development were flushed from the reproductive tract (control groups) of Wistar rats. Zygotes were flushed and grown in vitro to the above mentioned developmental stages and comprised the experimental groups. Immunofluorescence microscopy and computerized image analysis were used to evaluate both qualitative (localization) and quantitative expression of plasminogen activators. RESULTS: uPA and tPA were found to be expressed in rat embryos throughout their preimplantation development, both in vivo and in vitro. While uPA was localized mainly in the cell cytoplasm, the tPA was detected mainly on cell surface and in the perivitelline space. In blastocysts, both in vivo and in vitro, uPA and tPA were localized in the trophectoderm cells. Total uPA content per embryo was higher in the in vivo as compared with the in vitro developed embryos at all stages measured. Blastocyst uPA content was significantly low as compared with the four-cell, eight-cell, and morula stages. Total tPA content was higher in embryos developed in vivo than those developed in vitro except for the 4-cell and 8-cell stages. CONCLUSION: In vitro embryo development leads to lower PAs expression in a stage dependent manner as compared with in vivo developing controls. The enzymes studied vary probably in the ratio of their active and inactive forms as there is no correlation between their content and the activity observed in our previous study. The localization of both PAs in the blastocysts' trophectoderm supports the assumption that PAs plays a role in the implantation process in rats. BioMed Central 2005-02-10 /pmc/articles/PMC549534/ /pubmed/15703084 http://dx.doi.org/10.1186/1477-7827-3-7 Text en Copyright © 2005 Aflalo et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Aflalo, Eliahu D
Sod-Moriah, Uriel A
Potashnik, Gad
Har-Vardi, Iris
Expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro
title Expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro
title_full Expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro
title_fullStr Expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro
title_full_unstemmed Expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro
title_short Expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro
title_sort expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC549534/
https://www.ncbi.nlm.nih.gov/pubmed/15703084
http://dx.doi.org/10.1186/1477-7827-3-7
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