Identification and localization of heparan sulphate proteoglycans in non-pregnant and pregnant human uterus.

Authors:
A. Hjelm Cluff, MD1, A. Malmström, PhD2, K. Barchan, Tech. Ass.2, G. David, MD, PhD3, G. Ekman-Ordeberg, MD, PhD1.

Departement:
Department of Woman and Child Health, Division for Obstetrics and Gynecology, Karolinska Institute/Karolinska Hospital, Department of Cell and Molecular Biology, Lund University, Center for Human genetiecs, University of Leuven, Belgium

Source:
Presented at the Society for Gynecologic Investigation 50th Annual Meeting March 26-30, 2003, Washington DC Volume 10, Number 2 (Supplement), February 2003

Abstract:
OBJECTIVE: From our earlier studies we know that a remodeling of the proteoglycans in human uterus occur throughout pregnancy, and during labor. The hypothesis of this investigation is that heparan sulfate proteoglycans play a pivotal role for establishing of normal labor. In this study the different types of heparan sulphate proteoglycans in non-pregnant and pregnant human uterine tissue are separated, identified and localized in the tissue.

METHODS: Biopsies from the isthmic part of human uterus were obtained from women undergoing hysterectomy on benign indications, and from women during cæsarean section. They were extracted using 4 M guanidine hydrochloride containing 1% of Triton followed by chromatography on columns of DEAE-cellulose. After removal of chondroitin/dermatan sulfate proteoglycans by digestion with chondroitinase ABC, the heparan sulfate proteoglycan were purified using ion exchange chromatography and precipitation with Alcian blue. The heparan sulfate proteoglycans were subjected to controlled degradation using heparitinase or deglycosylation with trifluoro-methane-sulfonic acid. Separation was performed by SDS-PAGE and agarose gel electrophoresis followed by Western blotting. The localization was determined by immunohistochemistry using specific antibodies.

RESULTS: Separation after deglycosylation indicated the presence of syndecan 3, being the dominant one, syndecan 1, glypican and a component migrating as syndecan 2 or 4.Western blots after agarose electrophoresis using specific antibodies indicated the presence of the same proteoglycans and in addition perlecan. Using Western blot showed weak indications that perlecan, glypican 1 and syndecan 4 increased, whereas syndecan 1, syndecan 2 and syndecan 3 showed a slight decrease during pregnancy. Immunohistochemistry also indicates changes in localization during pregnancy.

DISCUSSION: The analyses show the presence and localization of several heparan sulfate proteoglycans and their possible variation during the remodeling of the uterine tissue during pregnancy.

Uppdaterad
2008-10-02

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