Abstract
The effect of heparin binding-epidermal growth factor (HB-EGF) on the in-vitro development of hamster 8-cell embryos was investigated. Supplementation of HB-EGF to culture medium accelerated zona escape of blastocysts (63 ± 9% compared with 33 ± 9% after 36 h; P < 0.05). Complete zona escape of blastocysts persisted even after 48 h (61 ± 11% versus 30 ± 4%) and 60 h (75 ± 6% versus 42 ± 8%). Addition of anti-HB-EGF antibody drastically reduced the percentage of zona escapedblastocysts (30.0 ± 5.0% versus 92.3 ± 2.8%; P < 0.05). Interestingly, a significant increase in the area of trophoblast outgrowth occurred in the presence of HB-EGF (116 × 103 ± 8 × 103 ìm2 versus 74 × 103 ± 8 × 103 ìm2 at 48 h; P < 0.05). This, however, was not due to an increased number of trophectodermal cells in HB-EGF-treated blastocysts. Immunoreactive HB-EGF was localized in blastocysts and uterine sections, visible by intense immunostaining in the luminal epithelium, particularly on the apical surface. Moreover, the expression of HB-EGF in the uterus was maximum on day 4 of pregnancy, coinciding with the timing of zona escape and implantation. The receptor of HB-EGF, viz. EGF receptor was also detected in blastocysts and the luminal epithelium of day 4 pregnant uterus. These results show that HB-EGF improves blastocyst hatching and trophoblast outgrowth in hamsters.
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