Hormonal profile and morphological changes in pig ovaries after intraovarian infusions of Escherichia coli endotoxin.

The purpose of this study was to estimate morphological changes in the ovary and size of the production of steroid hormones during the luteal phase of the estrous cycle in pigs after intraovarian infusions of Escherichia coli endotoxin. Polish Large White gilts (n = 15) of similar age (7-8 months) and body weight (90-110 kg) with two controlled subsequent estrous cycles were used. The animals were randomly divided into two groups: control (n = 9, the 10th day of the estrous cycle,) and treated with Escherichia coli endotoxin (n = 6, the same day of the estrous cycle). The gilts were infused with Escherichia coli endotoxin at a dose of 1 mg three times a day during six consecutive days, from the 14th to the 19th day of the estrous cycle. Plasma concentrations of progesterone (P4), androstenedione (A4), testosterone (T), estrone (E1) and estradiol-17 beta (E2) were determined by radioimmunoassay method. Infusions of Escherichia coli endotoxin resulted in a significant (p < 0.001) decrease in the production of P4, A4, T, E1 and E2 in the luteal phase as compared with the levels found in the control animals. Plasma level of P4, A4 and T was decreased by 84.6%, 86.0% and 73.0%, respectively. Plasma concentrations of E1 and E2 in some cases exceeded 5 pg/ml, nevertheless in the majority of the samples they were under sensitivity of the method. Escherichia coli endotoxin infusions resulted in a considerable decrease in the size of the ovaries, and morphological changes characteristic for acute and chronic inflammation were observed.

Effect of the gonadotrophins treatment on morphological alterations in ovary and peripheral plasma concentrations of steroid hormones in gilts.

The present study was designed to examine the influence of gonadotrophins treatment on the ovarian morphology changes and plasma concentrations of steroid hormones in peripheral blood. The experiment was performed on sexually pubertal gilts (Large White x Landrace) of similar age (7-8 months) and body mass (100-110 kg) with two controlled subsequent estrous cycles. The animals were randomly divided into four groups: two control consisting of pigs with the luteal phase (n = 9, the 10th day of the estrous cycle) and the follicular phase (n = 6, the 20th day of the estrous cycle) and two experimental ones consisting of animals with both mentioned periods (n = 7 and n = 9) treated with gonadotrophins (PMSG and hCG). The gilts in the luteal phase were injected (s.c.) with gonadotrophins at a daily dose of PMSG 400 and hCG 200 IU from the 16th to the 27th day (the 6th day of the next estrous cycle). The gilts in the follicular phase, were injected with the same dose of gonadotrophins but from the 8th to the 19th day of the estrous cycle. Plasma concentrations of P4, A4, T, E1, E2 and metabolite of PGF2 alpha-PGFM were determined by radioimmunoassay (RIA) method. Injections of PMSG and hCG in both experimental groups produced several times enlarged: weight, size and volume of ovaries and alterations in a number of structural elements as compared with those found in the control animals. The morphological elements presented in ovaries: corpora haemorrhagica, corpora lutea, regular and atretic follicles and first of all cysts by distinctly differentiation thickness of the walls are characteristic for cystic ovarian degeneration. Plasma concentrations all determined hormones after gonadotrophins treatment in experimental groups were increased except E1 (insignificant decrease) in luteal phase as compared with those found in the control groups. Statistically significant increase (p < 0.001) in plasma concentrations of P4, A4, and T in both experimental groups and E2 (p < 0.001) in luteal phase were noted. In peripheral plasma concentrations increase of E1 and E2 in follicular phase of the estrous cycle were insignificant.

Morphology of the dorsal nasal, frontal and facial veins in adult gilts.

Morphological examinations conducted on adult gilts indicate that the dorsal nasal, frontal and facial veins belong to the myoelastic type with a well-developed internal elastic lamina and a thick tunica media with elastic fibers. Smooth muscle cells in the tunica media are mainly arranged circularly. A characteristic feature of individual veins, and even their parts, was the difference in the distribution and number of elastic fibers and amount of collagen, both in the internal elastic lamina and tunica media. Slight thickening of the vessel walls and a decrease in the number of elastic fibers were observed in the distal part of the dorsal nasal vein and in the proximal parts of the frontal and facial veins. No valves were found in the frontal vein. The bundles of smooth muscle cells in the tunica media and elastic fibers surrounding them were rounded, not laminar like in the other veins. Characteristic, sandwich-like arranged smooth muscle bundles, elastic fibers and large amounts of collagen were observed in the tunica media of the distal part of the facial vein. A distinctive feature of the middle auricular and radial veins was the presence of a well-visible external elastic lamina in the adventitia. An evaluation of the luminous vein surface in a SEM shows that endothelial cells are elongated and arranged consistently with the direction of blood flow in almost all of the veins analyzed. Endothelial cells were less elongated in the distal part of the facial vein, and microvilli were present on them.

Light and scanning electron microscopy of the porcine mesometrial and paraovarian lymphatic networks.

Uterine lymphatics were examined in 38 pigs using light microscopy, stereomicroscopy, and scanning electron microscopy. Uterine lymphatics were initially filled with plastic materials: Microfil for stereomicroscopy, Mercox-corrosion cast for scanning electron microscopy (SEM), and Micropaque for x-ray photographs. Lymph precollectors of the uterine horns formed two superficial layers, ventral and dorsal. At the level of the ovary, precollectors joined to become collector lymph vessels, which entered nearby lymph nodes. Among the lymph vessels emanating from the uterus, there was a characteristics band of lymphatics that bordered on the isthmus of the oviduct. These passed toward the ovary to form the paraovarian lymphatic plexus. Segments of collector lymphatics were longer than precollectors, had thicker walls consisting of endothelial cells, smooth muscle (uniformly forming a continuous band around lymphatics) and fibroblasts. Both precollector and collector lymphatics were covered and surrounded by a fine network of blood microvessels (vasa-vasorum) especially well seen in corrosion casts (SEM).

Effect of estradiol and progesterone on growth of porcine myometrial smooth muscle cells and phospholipase C and adenylate cyclase signalling systems in vitro.

The objective of presented studies was to investigate whether estradiol and progesterone administered in vivo and/or added in vitro can influence the primary myometrial cell culture and how these steroid hormones can affect hCG stimulated cAMP and inositol phosphate production in the porcine uterine myocytes. Myometrial smooth muscle cells were obtained from six ovariectomized gilts pretreated (n = 4) or not (n = 2) with oestradiol benzoate and progesterone for 5 consecutive days. Immunocytochemical staining proved that the pattern of filamentous actin in the cytoplasm of the myometrial fibroblasts (basketlike network) was different from that of myometrial smooth muscle cells (long parallel fibres). The myocytes derived from steroid treated pigs and supplemented with estradiol and progesterone in vitro formed a hillock pattern on days 4-5 day of culture whereas cells obtained from not steroid pretreated gilts were smaller and did not create confluent form. Myocytes were treated in vitro with two doses of estradiol/progesterone (low - 0.2 nM/50 nM and high - 2 nM/500 nM, respectively) and two doses of hCG - 0.1 mU and 1000 mU/ml to study hCG action on the second messenger system in myocytes. The myometrial smooth muscle cells treated with low dose of estradiol and progesterone in vivo responded with much higher accumulation of inositol phosphates to strong (1000 mU/ml) hCG stimulation when compared with those receiving high dose of both steroids. The different doses of estradiol and progesterone caused a similar increase in basal cAMP accumulation as compared to control cells cultured without steroid hormones. hCG (0.1 mU/ml) had usually the additive effect on cAMP production in porcine myometrial cells. The presented paper shows that estradiol and progesterone administration in vivo followed by steroid hormone treatment in vitro affects the primary myometrial cells culture and that both steroid hormones modify the basal accumulation of the second messengers: cAMP and IP3 and their answer to hCG stimuli in pigs.

Vasa vasorum of blood and lymph vessels in the broad ligament of the sheep uterus analyzed by scanning electron microscopy.

The morphology of vasa vasorum of the blood and lymph vessels in the broad ligament of the uterus was examined in 12 adult ewes. The technique of vascular corrosion casts with the usage of Mercox resin allows multiple surveys of a three-dimensional organization of vascular networks under a scanning electron microscope (SEM). The study was appended by tissue pieces dried by the critical point method for SEM and semi-thin sections stained with toluidyne blue for light microscopy. Vasa vasorum surrounded both the larger vessels and the lymph vessels emanating from the ovary and the uterus. Significant differences in location and intensity of vasa vasorum in blood vessels were observed. They were less developed in the first part of the ovarian and the uterine arteries than in their smaller branches. Significantly large, two- or even three-layer vasa vasorum surrounded the ovarian and the uterine branches of the ovarian artery as well as the ovarian veins in the area of mesovarium. In the other areas of the broad ligament of the uterus, arterial vessels of similar sizes usually had a single-layer vasa vasorum. The lymph vessels leaving the ovary and uterus were surrounded by dense, although usually single-layed, microvascular network. The presence of vasa vasorum on relatively small blood and lymph vessels of the broad ligament of uterus indicates their particular functions that are probably connected with the estrous cycle or pregnancy and changes of blood flow rate entering the ovary and uterus. However, the presence of significantly developed, multi-layer vasa vasorum networks on only some of the blood vessels indicates that it is probably one of the vessels morphological adaptations to serve the counter-current transfer of biologically-active regulators in the area of the broad ligament of the sheep's uterus.

Differentiation of lymphatics from blood vessels in the broad ligament of the swine using S-100 protein immunohistochemical localization in the vascular endothelium.

We examined the immunohistochemical staining characteristics of S-100 protein in the vessels of the broad ligament of the swine uterus. The endothelial cells of arterial vessels, lymphatics and blood capillaries as well as nerve fiber bundles showed S-100 protein positivity. In contrast, the endothelial cells of veins did not react for the S-100 antiserum. Immunoreactivity for S-100 protein in the endothelial cells of lymphatics did not consistently demonstrate strong staining intensity. Accordingly, we filled lymphatics with colored gelatin before immunohistochemical staining to facilitate identification of lymphatics under light microscopy. Numerous arterioles and capillaries (of which the endothelial cells were immunopositive for S-100 protein) in the lymphatic walls, especially those in the paraovarian vascular plexus, support the existence of a microvascular arterio-arterial rete mirabile network.

Distribution and cycle phase dependency of gonadotropin receptors in musculature and blood vessels of the porcine broad ligament.

Luteinizing hormone (LH) and human chorionic gonadotropin (hCG) share common receptors in the uterus of many species. The aim of the present work was to investigate the possibility of LH/hCG binding sites present in the musculature of the porcine broad ligament, and to study the topography of LH/hCG receptors in ligamental blood vessels. Uteri of cyclic pigs were collected during follicular (n = 6) and midluteal (n = 6) phases of the estrous cycle. The horns and broad ligaments were dissected for measurements of numbers and affinities of unoccupied LH/hCG binding sites in crude cell membrane fractions of the myometrium and the smooth muscle layers of the ligament. The tissue samples for contact autoradiography, on film using [125I]hCG as ligand, were collected from different regions of the reproductive tract including: ovary, uterine horn, paraovarian vascular plexus, uterine vein, and uterine artery with supplying vessels and the outer arches of the broad ligament. The kidney and liver were used as negative control tissue. The specific binding of [125I]-labelled hCG in blood vessels was quantified on the base of optical density of the film. Scatchard analysis of data from broad ligamental and myometrial crude membrane preparations gave comparable estimates of the association constants and numbers of LH/hCG receptors. Mean concentrations (+/- SEM) of receptors (nmol/kg protein) were lower in the broad ligament than in the myometrium (p < 0.05) during both follicular and luteal phases (2.2 +/- 0.5 vs. 1.0 +/- 0.3 and 2.5 +/- 0.5 vs 1.7 +/- 0.2; respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Lymphatic vessels in the broad ligament of the uterus in swine.

The lymphatics of the broad ligament were depicted in 24 pigs by multiple interstitial injections of latex into the ovary and subserosa of the mesometrial margin of the uterine horn. Two morphologically different networks of lymphatics emanating from the uterus and ovary were established. Ovarian lymphatics leave the hilus, invariably enter the parovarian plexus, lie superficially under the perimetrium of the mesovarium, and run a parallel course closely intertwined with the blood vessels in this region. Some lymphatics are thus located close to the branches of the ovarian artery and the utero-ovarian vein. Whereas the ovarian and uterine lymphatics in the swine are not directly connected, the uterine lymphatics enter the mesovarium and lie in close proximity to both the ovarian lymphatics and nearby blood vessels.

Investigations of the course of post-partum period in the merino Polish sheep. IV. Shifts in the intercaruncular regions of uterus (observation in light microscope--LM and in scanning electron microscope--SEM).

Studies on the intercaruncular regions of uterus were carried out on 34 ewes on Days 1-78 of the post-partum period to determine the approximate date of ending post-partum changes and the appearance of morphological sings interrelated with the ovarian cycle phases. It was demonstrated that in sheep the remodelling of endometrium relief lasted until Day 12 of post-partum (p.p.) and at the same time destruction changes in the superficial and glandular epithelium, as well as in the uterine stroma ended. At first, changes in the myometrium (around Day 6--in the longitudinal layer and on Day 10--in the circular layer) were finished. After Day 12 p.p. there was a number of morphological signs indicating the relationship between the ovarian cycle phase and changes in the endometrium of intercaruncular regions of the uterus. At the same time, the role of stromal leucocytes and certain hormones in processes of the involution of uterus was discussed.

[Studies of the course of the postpartum period in sheep of the Polish Merino breed. III. Progesterone level in the peripheral blood 1-40 days after labor]. / Badania nad przebiegiem okresu poporodowego u owiec rasy merynos polski. III. Poziom progesteronu we krwi obwodowej w dniach od 1 do 40 po porodzie.

Progesterone concentration was determined by RIA in 24 ewes of Polish Merino Breed on 1-40th day after parturition. In most of the studied ewes (58%) 1-3 luteal phases were observed. Elevated concentration of progesterone appeared on days 6-15, but in most cases on the 20th day postpartum approximately. Luteal phases of first oestrous cycles were shortened and lasted approximately 6 days, whereas normal length of oestrous cycles and progesterone concentration illustrating the presence of cyclic corpus luteum were observed on the 25th day after postpartum.

[Studies of the course of the puerperal period in selected group of Polish Merino sheep. II. Clinical observations of the genital organs]. / Badania nad przebiegiem okresu poporodowego u wybranej grupy owiec rasy merynos polski. II. Obserwacje kliniczne narzadu plciowego.

Studies were carried out on 33 Polish Merino sheep. They concerned macro-morphological changes in the sexual organs during the postpartum period. Observations were made from the first to the 41st day postpartum. Changes in the sexual organs were described, as follows: skin swelling and colouring around perineum and vulvar lips, shape and size of the vaginal part of the uterine cervix, colour of the mucosa in the vaginal part of the uterine cervix, degree of opening of the external orifice of the uterine cervix, amount, consistency and colour of the postpartum discharges and uterine mucus. Observations revealed that in the examined group of ewes of the Polish Merino sheep the genital tracts became totally clean of the postpartum discharges, the mucosa of the vaginal part of the uterine cervix and of the vagina regained its normal colour by the 21st day of postpartum. The external orifice of the uterine cervix and the cervical canal also closed completely by this time. On the other hand, changes as to the shape and size of the vaginal part of the uterine cervix persisted as long as 33 days postpartum.

[Studies of the course of the postpartum period in Polish Merino sheep.I. Macromorphological studies of the ovary and uterus]. / Badania nad przebiegiem okresu poporodowego u owiec rasy merynos polski. I. Obserwacje makromorfologiczne jajnika i macicy.

The aim of the conducted research was to establish postpartum oestrous cycles and approximate data for mating Polish Merino sheep. 34 sheep were examined. During sheep laparotomy observations of ovaries and uterus were made on definite days of the postpartum period (1-78). It was stated that after about one-week of postpartum inactivity, the growth and development of follicles occurred in ovaries. First ovulations were noticed from the 15th to 18th day of postpartum, while processes connected with uterus involution were not completed. From the 14th to 18th day of postpartum, casting off of pregnant tissues from caruncules' surfaces into uterus cavity occurred. In the majority of sheep, regeneration of caruncules and purification of uterus cavity were completed by the 30th day of postpartum. The observed compatibility of the phases of ovary cycle with macromorphologically determined functional state of uterus after 30 days of postpartum, in the majority of tested sheep, suggests possibility of successful mating of Polish Merino sheep.