Expression of the epidermal growth factor receptor gene is regulated in mouse blastocysts during delayed implantation.

Publisher Website
Google Scholar
Abstract
The delayed implantation model was used to study epidermal growth factor receptor(s) (EGF-R) in the mouse blastocyst. Delayed implantation and blastocyst dormancy were induced by ovariectomy on day 4 of pregnancy and were maintained by daily (days 5-7) injections of progesterone (P4). Blastocyst activation and implantation were initiated by coinjection of estradiol-17 beta (E2) with P4 on the 3rd day of delay. Immunostaining of EGF-R, autoradiographic detection of 125I-labeled EGF binding, and measurement of EGF-inducible subcellular protein tyrosine phosphorylation demonstrated the loss of EGF-R from blastocysts (dormant) recovered 24 h after ovariectomy or on the 3rd day of P4-maintained delay. However, increased EGF-R levels were detected in blastocysts (activated) recovered 12 or 24 h after E2 injection. Blastocyst EGF-R mRNA levels were quantitated by reverse transcriptase (RT)-PCR and distribution of this mRNA was examined by in situ hybridization. To provide a homologous probe for these studies, a mouse EGF-R partial cDNA was cloned and used as the template for synthesis of antisense- and sense-strand EGF-R RNA. Quantitative RT-PCR demonstrated an 8- to 10-fold reduction in EGF-R mRNA copies per cell in dormant blastocysts. In contrast, an 8-fold increase in EGF-R mRNA copies per cell was detected in activated blastocysts 8 h after injection of E2. In situ hybridization detected EGF-R mRNA in most cells of normal day 4 blastocysts but not in those of dormant blastocysts. These studies establish that expression of the EGF-R gene in mouse blastocysts is tightly regulated by maternal steroid hormonal status.