Changes in intraconceptus pressure in the rabbit during days 7-10 of pregnancy

The present study established the magnitude of and temporal changes in the pressure within in situ rabbit conceptuses during early pregnancy. At the 7th, 8th, 9th, or 10th day post coitum (dpc), animals were anesthetized, and the number and dimensions of implantation sites were recorded. Conceptus pressure was measured by the servo-nulling method employing a glass micropipette. The mean number of sites was 6/cornu; spacing appeared normal. Implantation dome volume increased from 7 through 10 dpc: 0.2 +/- 0.03; 0.5 +/- 0.1; 1.6 +/- 0.2, and 2.7 +/- 0.9 cm3, respectively (Day 7 vs. 8, 9 and 10, p less than 0.01). Intraconceptus pressure declined between 7 and 10 dpc: 6.3 +/- 0.4; 5.7 +/- 0.4; 4.5 +/- 0.9, and 3.9 +/- 0.7 mm Hg, respectively (Day 7 vs. 10, p less than 0.05). Pressure fluctuated; the frequency of change in pressure varied between 7 and 8 dpc (4.2 +/- 3.0 to 2.3 +/- 3.5 peaks/min). The amplitude of fluctuation did not vary significantly between 7 and 10 dpc (2.6 +/- 0.3; 0.79 +/- 0.8 mm Hg, respectively). Simultaneous measurement of pressure within the conceptus and within the adjacent uterine lumen indicated that alterations in luminal pressure only occasionally influenced pressure within the conceptus. The decline in conceptus pressure suggests that the uterine wall becomes progressively more compliant as blastocyst cavity/yolk sac fluid accumulates within the conceptus. Conceptus expansion resulting from internal pressure may enhance conceptus-uterine metabolic exchange by facilitating apposition of conceptus-uterine surfaces and by increasing the ratio of conceptus surface to cytoplasmic mass.     

Artefakte im Amylase-Substratfilmtest

Zusammenfassung Bei der Verwendung von perjodsäureoxydierten Stärkefilmen (Shear und Pearse, 1963) zum Amylasenachweis können Artefakte entstehen durch alkalische, nicht-enzymatische Hydrolyse in Geweben mit hohem pH. Dies wird anhand von Beobachtungen an Kaninchen-Blastozysten erläutert.

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Artifacts in the substrate film test for amylase

Summary In substrate film tests for amylase, periodate-oxidized starch films (Shear and Pearse, 1963) may produce artifacts, for they undergo spontaneous hydrolysis at slightly alkaline pH levels. This is concluded from observations made on rabbit blastocysts.

     

Embryo implantation and proteinase activities in a marsupial (Macropus eugenii)

Summary Embryo implantation remains superficial (epithelio-chorial type) in most marsupials including the Macropodidae, but does involve formation of specialized contact zones of the trophoblast with the uterine epithelium. Since in eutherian mammals proteinases appear to play a central role in implantation-initiation mechanisms, a systematic histochemical investigation of proteinase patterns as related to implantation was performed in the tammar wallaby, Macropus eugenii (Macropodidae).Tammar uteri with embryos were collected at diapause and at days 7, 17, 18, 19, 20, 21 and 26 of the 27-day gestational period. Proteinase patterns were studied using a sensitive histochemical gelatin-substrate-film test previously optimized for the detection of trophoblast-dependent proteinase (blastolemmase) in the rabbit. Proteinase patterns were correlated with light-microscopical morphology of the processes of shedding of the extracellular embryo coverings (shell membrane) and attachment of the trophoblast to the uterine epithelium.At acid pH values an intracellular proteinase is detected in yolk sac endoderm and trophoblast as well as in endometrial glands and certain stromal cells. This enzyme is proposed to be a cathepsin indicating high catabolic activity connected particularly with protein transport from the endometrium into the yolk sac. Peak activity is found in the avascular (bilaminar) yolk sac at the phase when contact with the endometrium is being established.A particularly interesting proteinase active at alkaline pH values is detected in the trophoblast-endoderm complex. This enzyme appears to be extruded into the interface between trophoblast and uterine epithelium where it shows maximal activity for only approximately one day, around day (18-)19, exclusively in the bilaminar (avascular) yolk sac. The activity is correlated with the process of shedding of the extracellular embryo coverings (shell membrane) and of subsequent attachment of the trophoblast to the uterine epithelium, in the bilaminar but not the trilaminar (vascular) yolk-sac region. This is the first report on an extracellular (alkaline) proteinase activity possibly serving a specific function in embryo implantation in a marsupial.Abreviations BYS bilaminar (avascular) yolk sac membrane = bilaminar omphalopleure - dp.c. days post coitum - d RPY days after removal of pouch young - TYS trilaminar (vascular) yolk sac membrane = trilaminar omphalopleure Preliminary reports on portions of these investigations were presented at the 14th Annual Meeting of the Society for the Study of Reproduction 1981 (Biol Reprod 24 Suppl 1, p 78 A, 1981) and at the 3. Arbeitstagung der Anatomischen Gesellschaft 1982 (Anat Anz 153, 268, 1983)     

Endometrial arylamidase activity in the guinea pig: changes during the oestrous cycle, decidualization and ovarian steroid hormone treatment.

1. As suggested by comparative studies done in various species, amino acid arylamidases (amino peptidases) may play a role in blastocyst implantation. 2. Histochemical studies of the guinea pig endometrium indicate that arylamidase increases in the stroma during pregnancy but is depleted in the vicinity of the blastocyst during implantation. 3. To further explore the possible significance of arylamidases in uterine function, endometrial arylamidase activity was measured in guinea pigs during the reproductive cycle, decidualization and after ovariectomy with and without estrogen (E) and/or progesterone (P) treatment. Arylamidase activity was maximal during pro-oestrus-oestrus (40.0 +/- 10.0 mu/mg protein). 4. Enzyme activity was markedly depleted in decidualized endometrial stroma (12.3 +/- 1.6, P less than 0.01); reduced by ovariectomy (20.5 +/- 2.7); and stimulated by E (29.2 +/- 1.2); P had little effect (21.9 +/- 3.5). 5. The physiological significance of modulation of endometrial arylamidase activity by steroid hormones is discussed.