STUDIES ON EARLY DEVELOPMENT AND IMPLANTATION IN CAT .2. IMPLANTATION - PROTEINASES

The light microscopical morphology and proteinase activities are studied during the first phases of implantation in the cat, i.e. at 12, 13 and 14 days post coitum (d.p.c.). Timed matings are used to obtain exact data on the time course of implantation events. Size and shape of the blastocysts and the topographical relationships between trophoblast, zona pellucida and endometrium are studied on cryostat sections. These observations indicate that the zona pellucida is being removed at 12 d p.c. by dissolution which starts at the abembryonic pole and lateral of the embryonic disc. Since the zona has, in spite of the considerable expansion of the blastocyst, a thickness of 8-10 μm at this stage, it must have undergone a process of swelling or material must have been added invisibly. Invasion of the trophoblast into the endometrium begins between 13 and 14 d p. c. and is fully under way at 14 d p. c. Widening of the glandular lumina in the neighborhood of the blastocysts at 12-13 d p.c. indicates an early preimplantation interaction between the blastocyst and the endometrium. Amino Acid Arylamidase (Aminopeptidase) activity is found, in histochemical tests, to be high in the trophoblast but low in the endometrium in all three investigated stages. Proteinase Activity is studied with a highly sensitive gelatin substrate film test. Moderate to medium activity is found in the trophoblast at 12-13 d p.c. Very high proteinase activity is present in the invasion zone at 14 d p.c. Experiments with a large number of specific proteinase inhibitors in vitro and preliminary investigation of pH dependence show that it is mainly due to a cathepsin-B-like endopeptidase. This enzyme can be traced to both the trophoblast and the uterine epithelium. The disintegrating zona pellucida shows, at 12 d p.c., only little gelatinolytic proteinase activity. A trypsin-like endopeptidase as described for the rabbit blastocyst could not be identified with certainty in the cat but there is some indication that it might be present at 12 d p.c. Considerable trypsin-like proteinase activity is found in scattered endometrial stroma cells at all stages. The possible physiological role of the described proteinases in implantation is discussed. © 1978 Springer-Verlag.