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. 2009 Sep;201(3):322.e1-6.
doi: 10.1016/j.ajog.2009.06.002. Epub 2009 Jul 24.

The role of transforming growth factor beta in cervical remodeling within the rat cervix

Tanya Dailey  1 , Huiling Ji, Vit Long, Edward K Chien
Affiliations

The role of transforming growth factor beta in cervical remodeling within the rat cervix

Tanya Dailey et al. Am J Obstet Gynecol. 2009 Sep.

Abstract

Objective: Transforming growth factor beta (TGFbeta) plays a central role in extracellular matrix remodeling. We hypothesized that TGFbeta signaling is involved in cervical remodeling. This study evaluated patterns within this signaling pathway.

Study design: The cervices of nonpregnant and timed pregnant rats were obtained. Messenger ribonucleic acid (mRNA) expression of TGFbeta1, TGFbeta receptor 1 (TbetaR1), TbetaR2, and TbetaR3 was evaluated. Four animals were euthanized for each time point. Western blotting was performed for protein expression. Phosphorylated mothers against decapentaplegic (Smad)-2 and -3 phosphorylation was assessed to evaluate TGFbeta activation.

Results: TGFbeta1 mRNA increased through day 21 and declined on day 22 (analysis of variance, P = .001). TbetaR1 expression was unchanged. TbetaR2 and TbetaR3 mRNA expression was similar to TGFbeta1. TbetaR3 protein expression was similar to mRNA. Smad2 phosphorylation paralleled changes in TbetaR3.

Conclusion: Components of the TGFbeta signaling pathway increase during pregnancy along with Smad2 activation. The decline on day 22 correlates with a transition to the ripening phase supporting a role in cervical remodeling.

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Figures

Figure 1
Figure 1. TGFβ1 mRNA Expression in Timed Pregnant Rat Cervix
RT-PCR results representing the relative mRNA expression of TGFβ1 in timed pregnant rat cervixes. Values were normalized to β Actin. Values represent mean values obtained from 4 animals in each group run in triplicate. Error bars represent standard error. The increase in expression was found to be significant by ANOVA with p= 0.001. NP-Non Pregnant
Figure 2
Figure 2. TβR2 mRNA Expression in Timed Pregnant Rat Cervix
RT-PCR results representing the relative mRNA expression of TβR2 in timed pregnant rat cervixes. Values were normalized to β Actin. Values represent mean values obtained from 4 animals in each group run in triplicate. Error bars represent standard error. The increase in expression was found to be significant by ANOVA with p= 0.006. NP-Non Pregnant
Figure 3
Figure 3. TβR3 Expression in Timed Pregnant Rat Cervix
The blue columns represent TβR3 mRNA expression in timed pregnant rats. The maroon columns depict the densitometric analysis of TβR3 protein expression as determined by Western blot. Values were normalized to β Actin for both protein and mRNA studies. Values represent mean values obtained from 4 animals in each group. Error bars represent standard error. Amounts are relative to β Actin. NP-Non Pregnant
Figure 4
Figure 4. Phosphorylated Smad 2 Expression
Densitometric analysis of phosphorylated Smad 2 protein expression as determined by Western blotting. Values were normalized to β Actin. Values represent mean values obtained from 4 animals in each group. Error bars represent the standard error. NP-Non Pregnant
Figure 5
Figure 5. Smad 2 Expression in Cervical Tissue
Smad2 expression in cervical tissue. Smad2 expression is depicted in non pregnant (left panel) and gestational day 20 rat cervical tissue. Brown DAB staining intensity is qualitatively increased in day 20 compared to non-pregnant stromal tissue. Staining was done in parallel to limit variability in technique. There is decreased staining in endocervical epithelial cells on day 20 vs non pregnant tissue. Tissue counterstained with methyl green pyronin.
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