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  • Title: Differential regulation of heparin-binding epidermal growth factor-like growth factor in the adult ovariectomized mouse uterus by progesterone and estrogen.
    Author: Wang XN, Das SK, Damm D, Klagsbrun M, Abraham JA, Dey SK.
    Journal: Endocrinology; 1994 Sep; 135(3):1264-71. PubMed ID: 8070372.
    Abstract:
    Expression of heparin-binding epidermal growth factor-like growth factor (HB-EGF) was studied in the adult ovariectomized mouse uterus in response to progesterone (P4) and/or 17 beta-estradiol (E2) using Northern blotting, in situ hybridization, and immunohistochemistry. A 2.5-kilobase transcript of HB-EGF messenger RNA (mRNA) was detected in total uterine RNA samples. Although low levels of this mRNA were detected in uterine samples of oil-treated ovariectomized mice (control), an injection of E2 promptly up-regulated the levels. The mRNA levels peaked at 2 h and returned to basal levels after 12 h. Injection of P4 alone did not influence the basal levels; however, coinjection of E2 with P4 caused a rapid, but transient, up-regulation of the mRNA. The levels peaked between 2-4 h and declined 6 h after the hormone injections. Coinjection of E2 with P4 after 1 day of P4 priming also resulted in peak levels of HB-EGF mRNA at 2 h; however, the levels were not sustained thereafter. Because P4 and E2 differentially regulate heterogeneous uterine cell types, in situ hybridization was performed to determine cell-specific expression of HB-EGF mRNA in the ovariectomized uterus before and after steroid treatments. In the oil-treated uterine sections, very low levels of autoradiographic signals were observed in the luminal epithelium. In contrast, an injection of E2 resulted in a marked accumulation of HB-EGF mRNA primarily in uterine epithelial cells within 2 h. Although specific hybridization signals could not be detected in any uterine cell types after P4 treatment, combined treatment with P4 and E2 resulted in an accumulation of HB-EGF mRNA in stromal cells. To determine whether uterine HB-EGF mRNA was translated, cellular distribution of HB-EGF protein was investigated by immunohistochemistry. In oil-treated uterine sections, an overall weak immunostaining was noted, whereas no staining could be detected in uterine sections after P4 treatment. In contrast, positive immunostaining was noted in epithelial cells after E2 treatment. Coinjection of P4 with E2 caused immunostaining in the stroma. These results are consistent with those of in situ hybridization. The present investigation establishes that in the adult ovariectomized mouse uterus, E2 regulates HB-EGF expression in the epithelium, whereas expression of HB-EGF in the stroma is regulated by P4 and E2.
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