Differences in proteinase digestibility of the zona pellucida of in vivo and in vitro derived porcine oocytes and embryos

Embryo transfer practicioners know very well that, in a variety of species, there are differences between in vitro- and in vivo-derived embryos. It is assumed that these differences are results of suboptimal in vitro conditions leading to cytoplasmic and nuclear imperfections that will result in decreased embryo viability. In the present investigation the resistance of the zona pellucida of in vivo- and ex vivo-derived porcine embryos to a proteolytic enzyme is addressed. Ovulated but unfertilized oocytes, in vitro and in vivo-derived embryos of various developmental stages were exposed to a 0.5% pronase solution. The zonae of ovulated oocytes and in vivo-derived embryos at various stages of development took much longer to be digested than zonae of comparable in vitro-stages. Residence of in vitro derived embryos in an oviduct, no matter whether excised or in situ, significantly increased the zona resistance to pronase digestion. Embryonal stages normally residing in the uterus exhibited a distinctly decreased zona resistance to pronase. The culture of IVF embryos in an excised oviduct brought about a 6- to 14-fold increase in time required for zona digestion. A 24 h residence of IVF-derived embryos in the oviduct of a live recipient resulted in a digestion time for the zona of, on average, 48 h versus 1.4 h in the controls. Individual differences were substantial. The observations suggest that there must be structural changes in the zona pellucida or some sort of protective coating deposited while ova or embryos reside in the oviduct supplying protection against the action of proteolytic enzymes.