In order to clarify the distribution and content of estrogen (ER) and progesteron receptors (PR) under changing hormonal influences within the various cell populations of the uterus (glandular and luminal endometrial epithelium, stroma, myometrium), immunohistochemical determinations using specific monoclonal antibodies were made. To correlate the immunohistochemical findings with peripheral hormone levels and specific tasks of the endometrium, 17β-estradiol and progesterone serum levels were measured and cell proliferation determined by use of BrdU-labelling-immunohistochemistry. At the subcellular level ER and PR were located exclusively in the cell nuclei of female rabbits, which were either immature and lacking any peripheral hormone levels or were pseudopregnant (d0–d8 p.hCG). In the immature rabbits a general faint ER and PR immunostaining was found. In addition to a general increase in ER and PR in all cell populations estrous rabbits (d0 p.hCG) showed a significant rise of ER in the epithelial cells and of PR in the myometrium. Within the epithelial cells and the myometrium the ER dropped heavily within a few days of pseudopregnancy. The PR, however, increased sharply during the first two days of pseudopregnancy and decreased gradually following d4 p.hCG. A close relationship was observed between the high PR content and the proliferation rate of the epithelial cells on d2 p.hCG. In spite of the more rapid decrease of ER compared with PR, the glandular epithelium retained positive immunostaining. In the stroma the ER and especially PR content did not change significantly during the course of pseudopregnancy suggesting that some of the well-known differentiation events in the luminal epithelium may be mediated by the stroma.

We describe a well-established approach for studying the parameters and mechanisms of synchronization or desynchronization between the maternal and embryonic systems before implantation. It is useful for inducing 'delayed secretion' of the endometrium by different endocrine interventions, which dissociate the endometrial transformation from its control by the corpus luteum. The technique has been achieved by means of direct progesterone antagonists which competitively bind to the progesterone receptor and, in turn, inhibit the physiological effects of progesterone. During the luteal phase, secretory protein patterns indicate the receptive stage of the endometrium. Evidence is presented to show that these patterns, analysed by electrophoresis and densitometry, define the time at which an embryo transfer is promising for implantation and establishment of pregnancy.

There is evidence from previous studies that progesterone antagonists (antigestagens) modify estrogen responses at endometrial and myometrial levels without having affinity to the estrogen receptor (ER). The purpose of the present study was to investigate the influence of the antigestagen onapristone (ZK 98 299) on the uterus in ovariectomized (OVX) estradiol (E2)-substituted rabbits (3.0-mu-g/animal.day). The animals were treated for 8 days with different doses of onapristone (3.0, 10.0, and 30.0 mg/animal.day, sc). Uterine growth was not influenced by onapristone compared to that in OVX E2-substituted controls. However, morphological (light microscopy, transmission electron microscopy) and morphometric criteria indicated that there was a significant dose-dependent inhibition of the estrogen-induced gland formation within the endometrium and degenerative changes in glandular epithelial cells. By contrast, there were morphological signs of activation of the endometrial stroma (proliferation, increased capillarization, and vascularization, edema) above the level of E2-treated animals. A dose-dependent increase in the concentration of uterine cytosolic ER, nuclear ER, and ER mRNA (ER mRNA) was measured in uterine homogenates after onapristone treatment compared to values in OVX E2-substituted controls. Immunocytochemical analysis of ER in uterine sections suggests that the increase in ER after onapristone treatment took place predominantly in the myometrium and surface epithelium. To examine whether the observed interference was mediated via the progesterone receptor (PR), E2-substituted rabbits were treated, in a separate experiment, with onapristone (10.0 mg/animal.day, sc) and various doses of progesterone (1.0, 3.0, and 10.0 mg/animal.day, sc). Progesterone reversed all onapristone-induced changes, indicating that the observed effects were mediated via the PR. The data indicate that the antigestagen onapristone interacts with estrogen action in the absence of the natural PR ligand. The increase in ER and ER mRNA concentrations after onapristone treatment in OVX E2-treated animals suggests that this antigestagen abolished an inhibitory action of the unoccupied PR on ER biosynthesis.

Auf dem Gebiet der extrakorporalen Befruchtung sind in den letzten 10 Jahren enorme Fortschritte erzielt worden. Einige Teilbereiche dieses Therapieverfahrens wie z. B. die hormonelle Stimulation, die Follikelpunktion oder die Vorbe-handlung der Spermatozoen, erfuhren einerseits eine weitgehende Standardisierung, andererseits aber auch eine technische Vereinfachung oder Optimierung. Eine Stagnation der technischmethodischen Entwicklung ist dagegen im Bereich der Oocytenkultur zu verzeichnen, deren Prinzip seit 10 Jahren praktisch nicht verändert wurde. Eine neue Methode der In-vitro-Fertilisation wurde 1987 von Ranoux beschrieben [4]. Bei diesem Verfahren der intravaginalen Kultur (IVC), werden Eizellen zusammen mit den präparierten Spermatozoen in einer Plastik-kapsel in der Vagina der Patientin inkubiert. In der vorliegenden Arbeit möchten wir über eigene Erfahrungen mit diesem neuen Verfahren berichten.

In this study, we present further experiences with the intravaginal culture (IVC) of oocytes and cleavage stages. IVC is a new technique for in-vitro fertilization, its principle consisting of fertilization of oocytes in an air-free plastic capsule which is placed into the maternal vagina. In a total of 45 patients, 22 were treated by IVC and 23 by a conventional in-vitro fertilization technique (IVF). The pregnancy rates for IVC and IVF amounted to 22.7% (5/22) and 17.4% (4/23), respectively. Our results indicate that IVC is a valuable new method for treatment of special cases of sterility. Furthermore, extracorporal stress factors such as light and low temperature can be minimized by IVC. We also consider the psychological factor for the patients as very important since the mother can actively participate in the early development of the conceptus.

We present in this study our first experience with intravaginal culture (IVC) of human oocytes as a modified technique for in-vitro fertilization. Up to 4 oocytes and washed, pre-incubated spermatozoa (final concentration: 0.1-0.2 x 10(5)/ml) are pipetted into a plastic capsule. The capsule is closed, avoiding any air inclusion. It is placed into the maternal vagina for up to 50 hours, held in place by a diaphragm. The first 15 patients treated by IVC achieved a fertilization rate of 58% (35/60). Following transfer of the cleavage stages, 3 intact clinical pregnancies were established: 2 in the group with tubal sterility factors and 1 in the group of patients with endometriosis. In the group with andrological causes of sterility, one beta-hCG-positive reaction was detected. However, no intact clinical pregnancy was achieved. Our preliminary results show IVC to be a promising alternative in particular cases of sterility treatment. In contrast to the conventional technique, IVC minimizes exposure of gametes to factors which may affect their early development, such as light or low temperature. Furthermore, there is an important psychological factor to be considered, as the patient actively participates in the incubation period of in-vitro fertilization.

We present in this study our first experience with intravaginal culture (IVC) of human oocytes as a modified technique for in-vitro fertilization. Up to 4 oocytes and washed, pre-incubated spermatozoa (final concentration: 0.1-0.2 x 10(5)/ml) are pipetted into a plastic capsule. The capsule is closed, avoiding any air inclusion. It is placed into the maternal vagina for up to 50 hours, held in place by a diaphragm. The first 15 patients treated by IVC achieved a fertilization rate of 58% (35/60). Following transfer of the cleavage stages, 3 intact clinical pregnancies were established: 2 in the group with tubal sterility factors and 1 in the group of patients with endometriosis. In the group with andrological causes of sterility, one beta-hCG-positive reaction was detected. However, no intact clinical pregnancy was achieved. Our preliminary results show IVC to be a promising alternative in particular cases of sterility treatment. In contrast to the conventional technique, IVC minimizes exposure of gametes to factors which may affect their early development, such as light or low temperature. Furthermore, there is an important psychological factor to be considered, as the patient actively participates in the incubation period of in-vitro fertilization.

Proteinase inhibitor activity of rabbit uterine fluid during preimplantation and in the course of pseudopregnancy was investigated to determine if uteroglobin itself is actually a proteinase inhibitor.

In mammals, the reproductive process depends at all stages on the coordinated functioning of several physiologically closed systems. In the genital tract itself these are the mucous membranes and muscular structures. Their functioning is dependent on very complex hormonal and nervous controls and also on the blood supply, which changes according to the situation. There is a lot of evidence that various hormones and drugs can have a direct and significant influence on the structure and function of the uterus. Since the ovaries and their regulating systems are also complex, it is understandable that widely varying drugs can also indirectly influence processes in the uterus.