Accéder au contenu
Merck

PARP1 during embryo implantation and its upregulation by oestradiol in mice.

Reproduction (Cambridge, England) (2014-02-12)
Anubha Joshi, Sahil Mahfooz, Vineet Kumar Maurya, Vijay Kumar, Chadchan Sangappa Basanna, Gurpreet Kaur, Kashif Hanif, Rajesh Kumar Jha
RÉSUMÉ

Pregnancy requires successful implantation of an embryo, which occurs during a restricted period defined as 'receptivity of the endometrium' and is influenced by the ovarian steroids progesterone and oestradiol. The role of poly(ADP-ribose)polymerase-1 (PARP1) in apoptosis is well established. However, it is also involved in cell differentiation, proliferation and tissue remodelling. Previous studies have described the presence of PARP in the uterus, but its exact role in embryo implantation is not yet elucidated. Hence, in this study, we studied the expression of PARP1 in the uterus during embryo implantation and decidualisation, and its regulation by ovarian steroids. Our results show upregulation of the native form of PARP1 (∼116 kDa) in the cytosolic and nuclear compartments of implantation and non-implantation sites at day 5 (0500 h), followed by downregulation at day 5 (1000 h), during the embryo implantation period. The transcript level of Parp1 was also augmented during day 5 (0500 h). Inhibition of PARP1 activity by the drug EB-47 decreased the number of embryo implantation sites and blastocysts at day 5 (1000 h). Further, cleavage of native PARP1 was due to the activity of caspase-3 during the peri-implantation stage (day 5 (0500 h)), and is also required for embryo implantation, as inhibition of its activity compromised blastocyst implantation. The native (∼116 kDa) and cleaved (∼89 kDa) forms of PARP1 were both elevated during decidualisation of the uterus. Furthermore, the expression level of PARP1 in the uterus was found to be under the control of the hormone oestrogen. Our results clearly demonstrate that PARP1 participates in the process of embryo implantation.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
Réactif TRI®, For processing tissues, cells cultured in monolayer or cell pellets
Sigma-Aldrich
Cocktail d′inhibiteurs de protéases 2, aqueous solution (dark coloration may develop upon storage, which does not affect the activity)
Sigma-Aldrich
Cocktail d′inhibiteurs de protéases SIGMAFAST, comprimés, sans EDTA, for use in purification of Histidine-tagged proteins
Sigma-Aldrich
HEPES, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
Acide éthylènediaminetétraacétique solution, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
HEPES solution, 1 M in H2O
Sigma-Aldrich
Anticorps monoclonal de souris anti-β-actine−peroxydase antibody produced in mouse, clone AC-15, purified from hybridoma cell culture
Sigma-Aldrich
Progesterone, powder, BioReagent, suitable for cell culture
SAFC
HEPES
Sigma-Aldrich
Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture
Sigma-Aldrich
Ethylenediaminetetraacetic acid, 99.995% trace metals basis
Sigma-Aldrich
Progesterone, ≥99%
Sigma-Aldrich
Ethylenediaminetetraacetic acid, BioUltra, anhydrous, ≥99% (titration)
Sigma-Aldrich
Acide éthylènediaminetétraacétique disodium salt solution, BioUltra, for molecular biology, pH 8.0, ~0.5 M in H2O
SAFC
HEPES
Sigma-Aldrich
HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
Sigma-Aldrich
Ethylenediaminetetraacetic acid, purified grade, ≥98.5%, powder
Sigma-Aldrich
HEPES, BioXtra, pH 5.0-6.5 (1 M in H2O), ≥99.5% (titration)