Effect of Matrix Metalloproteinases-2 and -9 during IVC-2 on the Development Competence and Gene Expression Profile of Bovine In Vitro-Produced Embryos

Abstract

Matrix Metalloproteinases (MMP)-2 and -9 are participated in embryo development, implantation, remodeling ofepithelial cell and ovulation. The objective of this study is to evaluate an impact of MMP2 and MMP9 on embryonicdevelopmental competence as well as gene expression profiles of in vitro-produced bovine embryos. After in vitrofertilization, embryos of all groups were transferred into IVC-2 medium treated with MMP2 and MMP9 to check theoptimum concentration on the basis of embryo development competence and cell numbers. The optimum concentrationsfor MMP2 and 9 were 1,200 ng/ml and 300 ng/ml. The blastocyst development competence was not different among1,200 ng/ml of MMP2 vs. 300 ng/ml of MMP9 vs. combined MMP2 + 9 vs. control groups (41.46 ± 10.66 vs. 37.73± 8.92 vs. 45.11 ± 11.41% vs. 41.59 ± 11.88, respectively). Furthermore, the developmental competences to hatchingand hatched blastocysts were not also different among the same groups (79.84 ± 12.63 vs. 83.3 ± 17.46 vs. 78.55± 14.48% vs. 72.02 ± 14.09). In addition, total cell number was significantly (p<0.05) greater in blastocyst treated withMMP9 300 ng/ml among all treatment groups. On the other hand, there was no significant difference of ICM vs. TEratio in all groups. The expression of five out of six genes (i.e., MMP2, MMP9, IFNt, SSLP1 and HNRNPA2B1)was different among the groups. The expression of IFNt and HNRNPA2B1 genes was significantly greater in MMP9(p<0.05), but there was no difference of MMP9 expression between MMP2 and MMP9 group (p>0.05). The normalizedexpression of MMP2 and SSLP1 was greater in MMP2 than other groups (p<0.05). In conclusion, MMPs treatmentduring IVC-2 medium was remarkably effected on blastocyst developmental competence and gene expression profilesthat are related to embryo quality and implantation.