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Ovarian steroids regulate tachykinin and tachykinin receptor gene expression in the mouse uterus.

Pinto FM, Pintado CO, Pennefather JN, Patak E, Candenas L - Reprod. Biol. Endocrinol. (2009)

Bottom Line: Estrogen increased Tac1 and Tacr1 mRNA after 3 h and decreased Tac1 and Tac4 expression after 24 h.Tac2 and Tacr3 mRNA levels were decreased by estrogen at both 3 and 24 h.Most effects of estrogen were also observed in animals treated with propylpyrazole triol.

View Article: PubMed Central - HTML - PubMed

Affiliation: Instituto de Investigaciones Químicas, CSIC, Sevilla, Spain. Francisco.pinto@iiq.csic.es

ABSTRACT

Background: In the mouse uterus, pregnancy is accompanied by changes in tachykinin and tachykinin receptor gene expression and in the uterotonic effects of endogenous tachykinins. In this study we have investigated whether changes in tachykinin expression and responses are a result of changes in ovarian steroid levels.

Methods: We quantified the mRNAs of tachykinins and tachykinin receptors in uteri from ovariectomized mice and studied their regulation in response to estrogen and progesterone using real-time quantitative RT-PCR. Early (3 h) and late (24 h) responses to estrogen were evaluated and the participation of the estrogen receptors (ER), ERalpha and ERbeta, was analyzed by treating mice with propylpyrazole triol, a selective ERalpha agonist, or diarylpropionitrile, a selective agonist of ERbeta.

Results: All genes encoding tachykinins (Tac1, Tac2 and Tac4) and tachykinin receptors (Tacr1, Tacr2 and Tacr3) were expressed in uteri from ovariectomized mice. Estrogen increased Tac1 and Tacr1 mRNA after 3 h and decreased Tac1 and Tac4 expression after 24 h. Tac2 and Tacr3 mRNA levels were decreased by estrogen at both 3 and 24 h. Most effects of estrogen were also observed in animals treated with propylpyrazole triol. Progesterone treatment increased the levels of Tac2.

Conclusion: These results show that the expression of tachykinins and their receptors in the mouse uterus is tightly and differentially regulated by ovarian steroids. Estrogen effects are mainly mediated by ERalpha supporting an essential role for this estrogen receptor in the regulation of the tachykinergic system in the mouse uterus.

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Real-time quantitative PCR analysis of A) Tacr1; B) Tacr2 and C) Tacr3 in uterine cDNA from mice treated with E2, propylpyrazole triol (PPT), diarylpropionitrile (DPN)or estren. Uteri were collected from ovariectomized mice untreated (control) or treated for 3 or 24 h with E2 (1 μg/mouse) PPT (75 μg/mouse) DPN (100 μg/mouse) estren (300 μg/mouse) or the corresponding vehicle. Values are expressed as the fold change of each target-gene expression relative to the geometric mean mRNA expression of 4 housekeeping genes. Each bar represents the mean of uterine cDNA samples from at least five different mice, with SEM shown by vertical lines. *P < 0.05, significant difference versus mRNA levels in uteri from ovariectomized mice treated for 3 h with vehicle; ♦P < 0.05, significant difference versus mRNA levels in uteri from ovariectomized mice treated for 24 h with vehicle; one-way ANOVA.
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Figure 3: Real-time quantitative PCR analysis of A) Tacr1; B) Tacr2 and C) Tacr3 in uterine cDNA from mice treated with E2, propylpyrazole triol (PPT), diarylpropionitrile (DPN)or estren. Uteri were collected from ovariectomized mice untreated (control) or treated for 3 or 24 h with E2 (1 μg/mouse) PPT (75 μg/mouse) DPN (100 μg/mouse) estren (300 μg/mouse) or the corresponding vehicle. Values are expressed as the fold change of each target-gene expression relative to the geometric mean mRNA expression of 4 housekeeping genes. Each bar represents the mean of uterine cDNA samples from at least five different mice, with SEM shown by vertical lines. *P < 0.05, significant difference versus mRNA levels in uteri from ovariectomized mice treated for 3 h with vehicle; ♦P < 0.05, significant difference versus mRNA levels in uteri from ovariectomized mice treated for 24 h with vehicle; one-way ANOVA.

Mentions: The transcripts for Tac1, Tac2, Tac4, Tacr1, Tacr2 and Tacr3 were detected in uteri from ovariectomized mice. In all experiments and with all target genes, mRNA expression values were similar in untreated and vehicle-treated mice (Fig. 1, Fig. 2, Fig. 3 and Fig. 4).


Ovarian steroids regulate tachykinin and tachykinin receptor gene expression in the mouse uterus.

Pinto FM, Pintado CO, Pennefather JN, Patak E, Candenas L - Reprod. Biol. Endocrinol. (2009)

Real-time quantitative PCR analysis of A) Tacr1; B) Tacr2 and C) Tacr3 in uterine cDNA from mice treated with E2, propylpyrazole triol (PPT), diarylpropionitrile (DPN)or estren. Uteri were collected from ovariectomized mice untreated (control) or treated for 3 or 24 h with E2 (1 μg/mouse) PPT (75 μg/mouse) DPN (100 μg/mouse) estren (300 μg/mouse) or the corresponding vehicle. Values are expressed as the fold change of each target-gene expression relative to the geometric mean mRNA expression of 4 housekeeping genes. Each bar represents the mean of uterine cDNA samples from at least five different mice, with SEM shown by vertical lines. *P < 0.05, significant difference versus mRNA levels in uteri from ovariectomized mice treated for 3 h with vehicle; ♦P < 0.05, significant difference versus mRNA levels in uteri from ovariectomized mice treated for 24 h with vehicle; one-way ANOVA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2724541&req=5

Figure 3: Real-time quantitative PCR analysis of A) Tacr1; B) Tacr2 and C) Tacr3 in uterine cDNA from mice treated with E2, propylpyrazole triol (PPT), diarylpropionitrile (DPN)or estren. Uteri were collected from ovariectomized mice untreated (control) or treated for 3 or 24 h with E2 (1 μg/mouse) PPT (75 μg/mouse) DPN (100 μg/mouse) estren (300 μg/mouse) or the corresponding vehicle. Values are expressed as the fold change of each target-gene expression relative to the geometric mean mRNA expression of 4 housekeeping genes. Each bar represents the mean of uterine cDNA samples from at least five different mice, with SEM shown by vertical lines. *P < 0.05, significant difference versus mRNA levels in uteri from ovariectomized mice treated for 3 h with vehicle; ♦P < 0.05, significant difference versus mRNA levels in uteri from ovariectomized mice treated for 24 h with vehicle; one-way ANOVA.
Mentions: The transcripts for Tac1, Tac2, Tac4, Tacr1, Tacr2 and Tacr3 were detected in uteri from ovariectomized mice. In all experiments and with all target genes, mRNA expression values were similar in untreated and vehicle-treated mice (Fig. 1, Fig. 2, Fig. 3 and Fig. 4).

Bottom Line: Estrogen increased Tac1 and Tacr1 mRNA after 3 h and decreased Tac1 and Tac4 expression after 24 h.Tac2 and Tacr3 mRNA levels were decreased by estrogen at both 3 and 24 h.Most effects of estrogen were also observed in animals treated with propylpyrazole triol.

View Article: PubMed Central - HTML - PubMed

Affiliation: Instituto de Investigaciones Químicas, CSIC, Sevilla, Spain. Francisco.pinto@iiq.csic.es

ABSTRACT

Background: In the mouse uterus, pregnancy is accompanied by changes in tachykinin and tachykinin receptor gene expression and in the uterotonic effects of endogenous tachykinins. In this study we have investigated whether changes in tachykinin expression and responses are a result of changes in ovarian steroid levels.

Methods: We quantified the mRNAs of tachykinins and tachykinin receptors in uteri from ovariectomized mice and studied their regulation in response to estrogen and progesterone using real-time quantitative RT-PCR. Early (3 h) and late (24 h) responses to estrogen were evaluated and the participation of the estrogen receptors (ER), ERalpha and ERbeta, was analyzed by treating mice with propylpyrazole triol, a selective ERalpha agonist, or diarylpropionitrile, a selective agonist of ERbeta.

Results: All genes encoding tachykinins (Tac1, Tac2 and Tac4) and tachykinin receptors (Tacr1, Tacr2 and Tacr3) were expressed in uteri from ovariectomized mice. Estrogen increased Tac1 and Tacr1 mRNA after 3 h and decreased Tac1 and Tac4 expression after 24 h. Tac2 and Tacr3 mRNA levels were decreased by estrogen at both 3 and 24 h. Most effects of estrogen were also observed in animals treated with propylpyrazole triol. Progesterone treatment increased the levels of Tac2.

Conclusion: These results show that the expression of tachykinins and their receptors in the mouse uterus is tightly and differentially regulated by ovarian steroids. Estrogen effects are mainly mediated by ERalpha supporting an essential role for this estrogen receptor in the regulation of the tachykinergic system in the mouse uterus.

Show MeSH