RESUMO
Rats were studied to determine if matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) were detectable in developing embryonic and uterine tissues during the implantation period; that is, Days 6-8 of pregnancy. Tissue extracts were studied by zymography, reverse zymography and Western blotting. Immunohistochemistry procedures using antibodies against MMPs and TIMPs were applied to tissue sections that passed through implantation sites. The MMP inhibitor, doxycycline, was injected intraluminally into uterine horns of pseudopregnant animals. MMP-2 and -9, and TIMP-2 were detected in gelatin and reverse zymographs, respectively, from extracts of implantation sites on each day studied. Western blots defined bands corresponding to MMP-1, but casein zymographs did not consistently show evidence of extractable MMP-1 or -3. In tissue sections the primary decidual tissue area stained consistently for TIMP-2 and on Days 7 and 8 this area also had positive staining for MMP-1 in close proximity to the implanting embryo. On Day 8 positive staining in the outer stromal tissue was detected using an antibody against MMP-9, and another MMP (possibly MMP-3) was localized exclusively in ectoplacental cone/trophoblastic cells. Intraluminal injection of doxycycline did not significantly prevent the growth of the uterine horns following surgical induction of pseudopregnancy on Day 5. The strong localization of TIMP-2 in the primary decidual tissue is similar to that reported previously for TIMP-3, indicating that these inhibitors have a role in decidualization, including the regulation of trophoblastic invasion.
Assuntos
Implantação do Embrião , Metaloproteinases da Matriz/análise , Inibidores Teciduais de Metaloproteinases/análise , Útero/enzimologia , Animais , Western Blotting , Decídua/enzimologia , Doxiciclina/farmacologia , Eletroforese , Inibidores Enzimáticos/farmacologia , Feminino , Imuno-Histoquímica , Metaloproteinase 1 da Matriz/análise , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 9 da Matriz/análise , Inibidores de Metaloproteinases de Matriz , Gravidez , Pseudogravidez , Ratos , Ratos Wistar , Fatores de Tempo , Inibidor Tecidual de Metaloproteinase-1/análise , Inibidor Tecidual de Metaloproteinase-2/análise , Inibidor Tecidual de Metaloproteinase-3/análiseRESUMO
Uteri of pregnant rats on Days 6, 7 and 8 of pregnancy were studied to determine the histochemical distribution of collagen types III and V and the incorporation of [3H]glycine into fibrillar collagens during the period of embryonic implantation. Types III and V had a similar distribution in the non-decidual stromal region and muscle layers in implantation sites. They were found to have very low levels in the primary decidual tissue on Day 6 and were not detected in developing decidual tissues on Days 7 or 8. Following injection of labelled glycine, collagen was extracted and the specific activity of the collagens determined by fluorography and 3H incorporated into the collagen bands in the gels. It was found that incorporation of label into both types I and III was similar (33.4+/-12.0 and 31.8+/-18.1 cpm microg-1 collagen respectively) but 3.5 times that of type V (7.7+/-5.3 cpm microg-1). These studies suggest that although fibrillar collagens are metabolized or redistributed in the growing decidual tissue, they are incorporated rapidly into the extracellular matrix during remodelling of the outer stroma and muscle tissues.