Vasoactive intestinal peptide influences hatching of ovine blastocysts

Expanded blastocysts collected from superovulated Sarda ewes were divided a t random into four groups for culture in a simple medium that does not supp ort blastocyst hatching (CZB) or a complex medium that is permissive to hat ching (TCM 199), with or without vasoactive intestinal peptide (VIP), a kno wn embryo mitogenic peptide. Plasminogen activator (PA) secretion after 24 h of culture, and the number of cells, diameter of blastocysts and hatching rate after 48 h of culture were compared. The results showed an increase i n hatching rate (78.6 v. 6.7%; P<0.01), diameter and number of cells (220.8 9 v. 210.44 <mu>m, P<0.01 and 246 v. 232, P<0.01 respectively) and caseinol ytic areas (1.33 v. 0.92 cm, P<0.01) of blastocysts cultured in TCM 199 com pared with those cultured in CZB. Supplementation of the culture media with VIP increased these parameters in CZB (P<0.01) and partially in TCM 199. I n particular, cell number, diameter and PA activity were significantly high er (P<0.01) after culture with VIP in both media. Immunoneutralization of e xogenous VIP in culture with anti-VIP antibody caused a decrease in the hat ching rate (P<0.01) of embryos cultured in medium with VIP, similar to the rate in unsupplemented CZB (P<0.01). These results suggest a receptor-media ted response. In immunohistochemical studies, VIP was shown to bind recepto rs in hatched blastocysts demonstrating the VIP-receptor interaction, and V IP receptors of approximately 150 kDa were revealed by electrophoretic stud ies. In conclusion, ovine preimplantation embryos exhibit VIP receptors, pr oviding a basis for a receptor-mediated influence of VIP in the hatching of ovine blastocysts.