Expression of Epidermal Growth Factor (EGF) and the EGF Receptor in the Porcine Oviduct1

Abstract

The production, secretion, and localization of epidermal growth factor (EGF) and the distribution of the EGF receptor (EGF-R) were examined in the isthmus (I) and ampulla (A) of the oviducts from cyclic (C) and early-pregnant (P) gilts. Sexually mature gilts (n = 20) were divided equally into two groups: C and P. P gilts were bred twice (at 0 and 24 h), and all gilts were killed 48 h after onset of estrus. After removal of reproductive tracts, oviducts were isolated, flushed, opened longitudinally, divided by anatomical region, cut into 1-3-mm3 pieces, and placed in Dulbecco's modified Eagle's Essential medium (DMEM: F-12 + ITS [insulin, 5 micrograms/ml; transferrin, 5 micrograms/ml; and selenious acid, 5 ng/ml] + antibiotic). Half the tissue and medium were immediately homogenized and centrifuged, and the supernatant was removed. The remaining tissue was cultured in the medium for 24 h at 37 degrees C and 5% CO2, then prepared similarly for analysis. EGF was measured in the supernatant by a heterologous RIA. Concentration of EGF was expressed as nanogram/milliliter of EGF per milligram of protein in wet tissue. EGF concentrations were present in both regions of the oviducts of C and P gilts. It was greater in I than in A tissues for both C (I = 16.21 ng/ml vs. A = 13.91 ng/ml; p < 0.05) and P gilts (I = 14.27 ng/ml vs. A = 12.53 ng/ml; p < 0.10). Higher concentrations of EGF were found in I tissue of C gilts than in P gilts (C = 16.21 ng/ml vs. P = 14.27 ng/ml; p < 0.05). The media assayed from cultured explants of I and A sections from C and P gilts gave results that were highly correlated with those of immediately prepared tissue sections. Localization of EGF in frozen oviductal tissue sections was demonstrated by immunohistochemistry. The primary site of EGF immunostaining occurred in the epithelial cells (with highest intensity at the apical border) of both C and P gilts. A and I tissue sections from C gilts showed localization of EGF immunostaining mainly in epithelial cells and lamina propria cells, while those from P gilts stained less intensely. The presence of EGF-R was shown by incubating tissue imprints and frozen sections with EGF-erythrosin isothiocyanate, which revealed that EGF-R were distributed mainly on the membranes of epithelial cells. The study indicates that EGF and EGF-R are present in oviductal epithelial cells in both C and P gilts, with the highest concentration of EGF in C gilts.(ABSTRACT TRUNCATED AT 250 WORDS)