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Hormone-regulated expression and distribution of versican in mouse uterine tissues.

Salgado RM, Capelo LP, Favaro RR, Glazier JD, Aplin JD, Zorn TM - Reprod. Biol. Endocrinol. (2009)

Bottom Line: This study analyzed the synthesis and distribution of VER in mouse uterine tissues during the estrous cycle, in ovariectomized (OVX) animals and after 17beta-estradiol (E2) and medroxyprogesterone (MPA) treatments, either alone or in combination.Real Time PCR analysis showed that VER expression increases considerably in the MPA-treated group.These results show that the expression of versican in uterine tissues is modulated by ovarian steroid hormones, in a tissue-specific manner.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Reproductive and Extracellular Matrix Biology, Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil. rsalgado@icb.usp.br

ABSTRACT

Background: Remodeling of the extracellular matrix is one of the most striking features observed in the uterus during the estrous cycle and after hormone replacement. Versican (VER) is a hyaluronan-binding proteoglycan that undergoes RNA alternative splicing, generating four distinct isoforms. This study analyzed the synthesis and distribution of VER in mouse uterine tissues during the estrous cycle, in ovariectomized (OVX) animals and after 17beta-estradiol (E2) and medroxyprogesterone (MPA) treatments, either alone or in combination.

Methods: Uteri from mice in all phases of the estrous cycle, and animals subjected to ovariectomy and hormone replacement were collected for immunoperoxidase staining for versican, as well as PCR and quantitative Real Time PCR.

Results: In diestrus and proestrus, VER was exclusively expressed in the endometrial stroma. In estrus and metaestrus, VER was present in both endometrial stroma and myometrium. In OVX mice, VER immunoreaction was abolished in all uterine tissues. VER expression was restored by E2, MPA and E2+MPA treatments. Real Time PCR analysis showed that VER expression increases considerably in the MPA-treated group. Analysis of mRNA identified isoforms V0, V1 and V3 in the mouse uterus.

Conclusion: These results show that the expression of versican in uterine tissues is modulated by ovarian steroid hormones, in a tissue-specific manner. VER is induced in the myometrium exclusively by E2, whereas MPA induces VER deposition only in the endometrial stroma.

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Related in: MedlinePlus

Relative mRNA expression of versican in the mouse uterus. The mRNA expression was analyzed by real-time PCR using primers common to all splice variants, and relative gene expression determined by designating estrus to 1. (A) versican mRNA in estrus and diestrus; (B) versican mRNA after ovariectomy and hormone replacement. Values represent the mean + SE of determinations on three independent tissue preparations. # p < 0.05 vs. estrus by Student t-test; *p < 0.05 vs. ovx and E2; ** p < 0.001 vs. all by ANOVA.
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Figure 3: Relative mRNA expression of versican in the mouse uterus. The mRNA expression was analyzed by real-time PCR using primers common to all splice variants, and relative gene expression determined by designating estrus to 1. (A) versican mRNA in estrus and diestrus; (B) versican mRNA after ovariectomy and hormone replacement. Values represent the mean + SE of determinations on three independent tissue preparations. # p < 0.05 vs. estrus by Student t-test; *p < 0.05 vs. ovx and E2; ** p < 0.001 vs. all by ANOVA.

Mentions: VER mRNA was estimated by real time PCR of uterine cDNA obtained during the estrous cycle or after ovariectomy or hormone replacement (n = 5). Figure 3A shows that the relative expression of VER was ~3 fold higher in estrus than in diestrus (p < 0.05). After ovariectomy, VER mRNA was increased in the MPA-treated group (~2 fold; p < 0.001). The treatment with E2 or E2+MPA did not significantly alter VER mRNA, if compared to estrus. However, oil injections significantly decreased VER mRNA expression when compared to Ovx and E2 (Figure 3B).


Hormone-regulated expression and distribution of versican in mouse uterine tissues.

Salgado RM, Capelo LP, Favaro RR, Glazier JD, Aplin JD, Zorn TM - Reprod. Biol. Endocrinol. (2009)

Relative mRNA expression of versican in the mouse uterus. The mRNA expression was analyzed by real-time PCR using primers common to all splice variants, and relative gene expression determined by designating estrus to 1. (A) versican mRNA in estrus and diestrus; (B) versican mRNA after ovariectomy and hormone replacement. Values represent the mean + SE of determinations on three independent tissue preparations. # p < 0.05 vs. estrus by Student t-test; *p < 0.05 vs. ovx and E2; ** p < 0.001 vs. all by ANOVA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2698856&req=5

Figure 3: Relative mRNA expression of versican in the mouse uterus. The mRNA expression was analyzed by real-time PCR using primers common to all splice variants, and relative gene expression determined by designating estrus to 1. (A) versican mRNA in estrus and diestrus; (B) versican mRNA after ovariectomy and hormone replacement. Values represent the mean + SE of determinations on three independent tissue preparations. # p < 0.05 vs. estrus by Student t-test; *p < 0.05 vs. ovx and E2; ** p < 0.001 vs. all by ANOVA.
Mentions: VER mRNA was estimated by real time PCR of uterine cDNA obtained during the estrous cycle or after ovariectomy or hormone replacement (n = 5). Figure 3A shows that the relative expression of VER was ~3 fold higher in estrus than in diestrus (p < 0.05). After ovariectomy, VER mRNA was increased in the MPA-treated group (~2 fold; p < 0.001). The treatment with E2 or E2+MPA did not significantly alter VER mRNA, if compared to estrus. However, oil injections significantly decreased VER mRNA expression when compared to Ovx and E2 (Figure 3B).

Bottom Line: This study analyzed the synthesis and distribution of VER in mouse uterine tissues during the estrous cycle, in ovariectomized (OVX) animals and after 17beta-estradiol (E2) and medroxyprogesterone (MPA) treatments, either alone or in combination.Real Time PCR analysis showed that VER expression increases considerably in the MPA-treated group.These results show that the expression of versican in uterine tissues is modulated by ovarian steroid hormones, in a tissue-specific manner.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Reproductive and Extracellular Matrix Biology, Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil. rsalgado@icb.usp.br

ABSTRACT

Background: Remodeling of the extracellular matrix is one of the most striking features observed in the uterus during the estrous cycle and after hormone replacement. Versican (VER) is a hyaluronan-binding proteoglycan that undergoes RNA alternative splicing, generating four distinct isoforms. This study analyzed the synthesis and distribution of VER in mouse uterine tissues during the estrous cycle, in ovariectomized (OVX) animals and after 17beta-estradiol (E2) and medroxyprogesterone (MPA) treatments, either alone or in combination.

Methods: Uteri from mice in all phases of the estrous cycle, and animals subjected to ovariectomy and hormone replacement were collected for immunoperoxidase staining for versican, as well as PCR and quantitative Real Time PCR.

Results: In diestrus and proestrus, VER was exclusively expressed in the endometrial stroma. In estrus and metaestrus, VER was present in both endometrial stroma and myometrium. In OVX mice, VER immunoreaction was abolished in all uterine tissues. VER expression was restored by E2, MPA and E2+MPA treatments. Real Time PCR analysis showed that VER expression increases considerably in the MPA-treated group. Analysis of mRNA identified isoforms V0, V1 and V3 in the mouse uterus.

Conclusion: These results show that the expression of versican in uterine tissues is modulated by ovarian steroid hormones, in a tissue-specific manner. VER is induced in the myometrium exclusively by E2, whereas MPA induces VER deposition only in the endometrial stroma.

Show MeSH
Related in: MedlinePlus