Search

Pig embryos enter the uterus from the oviduct about 48 h after ovulation at about the 4-cell stage. They remain near the tip of the uterine horn until about Day 6 when they migrate towards the body of the uterus. By Day 9 some embryos have entered the horn opposite the one of origin and continue migrating until Day 12. At Day 12 embryos can no longer successfully move to a different site. The critical signal for recognition of pregnancy occurs at Day 12 at the same time that embryos are elongating at a rapid rate. The distribution of embryos throughout the length of the horns has occurred by this time and spacing accompanied by uterine growth takes place. The speed of migration is not influenced by the number of embryos but may be affected by secretions from the embryos acting on the uterus. A significant section of unoccupied uterus at Day 12 will prevent continuance of pregnancy regardless of the number of embryos present in the occupied section. When the number of embryos is so few as to not occupy the uterus fully, pregnancy will not continue. Removal of embryos from a significant section of the uterus after Day 14 does not stop an existing pregnancy. Embryos are spaced nearly equidistant from each other with the absolute space between fetuses dependent on the total uterine space available. The position of the fetus in the uterus affects the spacing between fetuses; fetuses at the tip of the horn before Day 25 have the greatest space with the space decreasing from the tip to the body.(ABSTRACT TRUNCATED AT 250 WORDS)

In pigs, transcervical infusion of seminal plasma at the onset of oestrus advances ovulation and thus improves the chance of gametes meeting during their full fertilizing competence. An animal model that allows single uterine horn infusion was used in combination with transcutaneous sonographic monitoring of ovaries. Preparative surgery involved the detachment of one uterine horn from the corpus, leaving the caudal end open to the peritoneal cavity but sealing the corpus wound. Transcervical infusion of 100 ml seminal plasma immediately after the detection of oestrus advanced ovulation by between 8 and 14 h on the ipsilateral ovary adjacent to the infused horn compared with the contralateral ovary. In addition, the seminal plasma infusion did not influence the LH profile compared with uterine infusion of PBS. This finding indicates that the effect is mediated by a local mechanism in the female genital tract. The advancement of ovulation depends on the time of infusion early in oestrus and is more pronounced in gilts exhibiting a long interval between the onset of oestrus and spontaneous ovulation compared with early ovulators. At 24 h after the detection of oestrus, seminal plasma was ineffective. Apparently, seminal plasma does not affect maturation and fertilizing competence of oocytes. The activity resides in a low molecular mass protein fraction and, to a far lesser extent, in seminal oestrogens. Further characterization of the active components may allow a practical application in AI.

Implantation/placentation in domestic pigs is preceded by synthesis of oestrogen by the conceptus to maintain functional corpora lutea throughout pregnancy and a rapid morphological transformation of conceptuses from spherical to long filamentous thread-like structures. Initial conceptus expansion, reaching a metre in length, not only delineates the surface area for placental attachment, but also provides the mechanism for delivery of oestrogen to signal events necessary for placentation throughout the uterine horn. Timing for conceptus gene expression to induce trophoblast expansion and attachment in pigs is temporally associated with downregulation of progesterone receptors and increase in oestrogen receptors within the uterine epithelium. Within the confines of the uterine lumen, pig conceptuses normally do not erode or invade through the uterine epithelial surface. However, the pig conceptus possesses extensive proteolytic activity as it is highly invasive outside the uterine lumen of the pig. Initial release of oestrogen by the elongating pig conceptus induces endometrial release of cytokines and a variety of protease inhibitors. Recently, endometrial expression for the inter-trypsin inhibitor (I alpha I) family of protease inhibitors has been detected in the pig endometrium during conceptus elongation and attachment. It is possible that I alpha Is may function to inhibit trophoblast invasion and also serve as targets for adhesion molecules, such as integrins and heparin, to aid in placental attachment to the uterine epithelium.

The aim of this study was to identify uterine pluripotent cells both in bovine uterine tissues as well in epithelial, stromal, and myometrial uterine cell populations. Moreover, the relationship of pluripotent markers expression with age and the uterine horn side was considered. Uterine tissue was collected from ipsilateral and contralateral horns (days 8–10 of the estrous cycle). Immunohistostaining for C-KIT, OCT3/4, NANOG, and SOX2 in uterine tissue was determined. mRNA expression of C-KIT, OCT3/4, NANOG and SOX2 was evaluated in uterine tissue relative to the age of the cow and uterine horn side. Gene and protein expression of these markers in the uterine luminal epithelial, stromal, and myometrial cells was evaluated by real-time PCR and western blotting respectively. The expression of pluripotent cell markers OCT3/4, NANOG, and SOX2 was identified by flow cytometry assay in epithelial, stromal, and myometrial cells. Multilineage differentiation of the bovine uterine cells was performed. mRNA expression of OCT3/4, NANOG, and SOX2 in uterine tissue was higher in the ipsilateral horn than in the contralateral horn. Flow cytometry assay revealed positive fluorescence for OCT3/4, NANOG, and SOX2 in all uterine cell types. Results showed the age-dependent expression of pluripotent markers in uterine tissue. Beside, the different expression of pluripotent cells in each horn of uterus suggests the influence of ovarian hormones on these characteristics. The highest mRNA and protein expression for pluripotent markers was observed in stromal cells among uterine cells, which indicates this population of cells as the main site of pluripotent cells in the cow uterus.

The fertility of ram semen after cervical insemination is substantially reduced by 24 h of storage in liquid form. The effects of liquid storage on the transit of ram spermatozoa in the ewe genital tract was investigated using a new procedure allowing direct observation of the spermatozoa in the genital tract. Ejaculated ram spermatozoa were double labeled with R18 and MitoTracker Green FM, and used to inseminate ewes in estrus either cervically through the vagina or laparoscopically into the base of the uterine horns. Four hours after insemination, the spermatozoa were directly observedin situusing fibered confocal fluorescence microscopy in the base, middle and tip of the uterine horns, the utero-tubal junction (UTJ) and the oviduct. The high resolution video images obtained with this technique allowed determination of the distribution of spermatozoa and individual motility in the lumen of the ewe's genital tract. The results showed a gradient of increasing concentration of spermatozoa from the base of the uterus to the UTJ 4 h after intra-uterine insemination into the base of the horns. The UTJ was shown to be a storage region for spermatozoa before their transfer to the oviduct. Thein vitrostorage of spermatozoa in liquid form decreased their migration through the cervix and reduced the proportion of motile spermatozoa and their straight line velocity at the UTJ and their transit into the oviduct.

In the present study, sperm distribution in the genital tract of the bitch following artificial insemination (AI) in relation to the time of ovulation was investigated by histology, scanning electron microscopy (SEM) and flushing. Ten bitches were inseminated intravaginally with 500 × 106spermatozoa: three dogs before ovulation, four dogs during ovulation and three dogs after ovulation. Ovariohysterectomy was performed 24 h after AI. Half of the genital tract was divided into nine segments (cervix, corpus uteri, caudal, middle and cranial uterine horn (UTH), utero–tubal junction (UTJ), isthmus, ampulla and infundibulum), which were processed for histology and SEM. The contralateral UTH and uterine tube (UT) were flushed, and several sperm characteristics were assessed. Histology revealed that the spermatozoa were mainly located in the uterine glands and at the UTJ, while very few spermatozoa were detected in the UT. Insemination during ovulation resulted in higher percentages of glands with spermatozoa in the different parts of the uterus (P< 0.05). Evaluation by SEM showed higher numbers of spermatozoa in several parts of the uterus for bitches inseminated during ovulation (P< 0.05). The mean number of spermatozoa flushed from the UTH and the UT was low. No significant differences in the evaluated sperm quality parameters were found between the flushings of the UTH and the UT. In conclusion, based on our findings, the uterine glands and the UTJ might act as sperm reservoirs in the bitch and sperm transport in the genital tract is affected by the time of AI in relation to ovulation.

First postpartum dominant follicles are preferentially selected in the ovary contralateral to the previously gravid uterine horn. The aim of the present study was to test the hypothesis that uterine bacterial contamination alters the location of ovarian follicle emergence and selection, and inhibits follicle growth and function. Swabs were collected from the uterine body lumen of cattle on days 7, 14, 21 and 28 after parturition. Bacteria were identified by aerobic and anaerobic culture; bacterial growth was scored semiquantitatively and animals were categorized into standard or high bacterial contamination categories on the basis of the number of colonies detected. Follicular growth and function were monitored by daily transrectal ultrasonography, and estimation of plasma FSH, oestradiol and progesterone concentrations. There was no effect of bacterial contamination on plasma FSH concentration profiles or emergence of the ovarian follicle wave. When uterine bacterial growth scores were high on day 7 or day 21 after parturition, fewer first (1/20 versus 15/50; P < 0.05) or second (1/11 versus 13/32; P < 0.05) dominant follicles were selected in the ipsilateral compared with the contralateral ovary, respectively. The diameter of the first dominant follicle was smaller in animals with a high day 7 bacterial score (P < 0.001), dominant follicle growth was slower (P < 0.05) and oestradiol secretion was decreased (P < 0.05). The present study provides evidence for an effect of the uterus on the ovary after parturition, whereby uterine bacteria have a contemporaneous localized effect on ovarian follicle selection and subsequent growth and function, but not on initial emergence.

The relationships between activities of matrix metalloproteinases (MMPs) in the canine uterus and the occurrence of degeneration of the luminal epithelium, cystic endometrial hyperplasia, pyometra and uterine remodelling post partum were determined. Mature bitches (n = 10) were ovariectomized, treated with hormones (oestradiol benzoate, progestagen) and investigated at stages simulating pro-oestrus (n = 2), oestrus (n = 2), dioestrus (n = 2), and mid- (n = 2) and late (n = 2) anoestrus (3 and 9 weeks, respectively, after cessation of treatment with progestagen). Untreated bitches (n = 1 per group) served as controls (Expt 1). An additional 10 ovariectomized bitches, at the end of treatment-induced simulated dioestrus, were treated each day for a further 3 weeks either with the same dose (standard dose, n = 3), a decreased dose (n = 3) or an increased dose (n = 3) of progestagen, or no treatment (withdrawal dose, n = 1). These bitches were then investigated (Expt 2). A suture was placed in the lumen of one uterine horn of another five bitches at ovariectomy. Three of these bitches were treated to induce simulated dioestrus and two bitches served as untreated controls. In the hormone-treated bitches, the suture resulted in cystic endometrial hyperplasia in one bitch and in cystic endometrial hyperplasia with pyometra in two bitches. The control bitches showed no cystic endometrial hyperplasia or pyometra (Expt 3). Four intact bitches were studied at 2 (n = 1), 3 (n = 2) and 11 (n = 1) weeks post partum. Uterine tissues were also collected from two bitches with naturally occurring cystic endometrial hyperplasia with pyometra (Expt 4). All uteri were examined histologically and the activities of MMP-2, -7 and -9 (latent and active forms) were assessed using zymography of extracts of endometrium. In Expts 1 and 2, marked degeneration of the luminal epithelium in six of 25 bitches (simulated mid-anoestrus, withdrawal dose and decreased dose groups) was not associated with changes in MMP activities. Markedly increased activities of MMP-2 (active form), -7 (latent form) and -9 (active and latent forms) were observed in the bitches with cystic endometrial hyperplasia with pyometra (but not with cystic endometrial hyperplasia alone) and in the bitches at 2 and 3 weeks post partum (but not at 11 weeks post partum). These results indicate that MMPs are not involved with degeneration of the luminal epithelium, but are involved with uterine remodelling in the postpartum bitch and with cystic endometrial hyperplasia with pyometra.

A fibreoptic endoscope procedure for non-surgical deep intrauterine insemination in non-sedated sows has been reported. However, the endoscope is an expensive and fragile instrument, and is unsuitable for use under field conditions. The aim of this study was to determine the minimum number of spermatozoa required to maintain optimal fertility using a flexible catheter (1.8 m in length, 4 mm in diameter) for deep intrauterine insemination in 2-6 parity non-sedated sows. Crossbred sows were treated with eCG 24 h after weaning and with hCG 72 h later to induce oestrus. Deep intrauterine insemination was performed 36 h after hCG treatment in 117, 126, 60 and 69 sows with 15.0, 5.0, 2.5 or 1.0 x 10(7) spermatozoa in 10 ml, respectively. Weaned sows (n = 147) not treated with hormones and used for standard artificial insemination (AI) (two inseminations per oestrus with 3 x 10(9) spermatozoa in 100 ml) served as controls. The flexible catheter was passed successfully through the cervix into one uterine horn in 95.4% of the sows in an average of 3.7 +/- 0.09 min. Farrowing rates after deep intrauterine insemination with 15 or 5 x 10(7) spermatozoa did not differ from those of the control group (82.9, 76.2 and 83.0%, respectively), but a significant decrease (P < 0.001) was observed in sows inseminated with 2.5 or 1.0 x 10(7) spermatozoa (46.7 and 39.1%, respectively). In contrast, the number of spermatozoa inseminated did not affect prolificacy. Laparotomy revealed that the tip of the flexible catheter reached approximately the anterior third of the uterine horn. Although deep intrauterine insemination was performed in only one uterine horn, the percentages of embryos collected from the tip of both uterine horns 2 days after deep insemination were not significantly different. The results show that in comparison with standard AI, a 20-60-fold reduction in the number of spermatozoa inseminated and an 8-10-fold reduction in the dose volume can be achieved without decreasing fertility when semen is deposited non-surgically into the upper first third of one uterine horn.

Experiments were performed to test the null hypotheses that embryonic survival is not affected by the presence of regressing corpora lutea in progestogen-supplemented ewes, and that the embryotoxic effects of regressing corpora lutea do not act locally on embryos in the uterine horn adjacent to the regressing corpora lutea. In Expt 1, laparotomies were performed on day 4 after mating, and progestogen supplementation was initiated and continued until pregnancy was diagnosed by ultrasonography on day 25. On day 4 after mating, ewes were lutectomized (n = 17) or sham lutectomized (n = 15), and injected (i.m.) with 5 mg PGF2alpha at 8 h intervals for 2 or 3 days. Controls (n = 14) were sham lutectomized and injected with saline as described above. Pregnancy rates did not differ in ewes treated with PGF2alpha for 2 rather than 3 days. Pregnancy rates were lower in ewes treated with PGF 2a compared with controls (P < 0.01). In ewes treated with PGF2alpha, lutectomy resulted in an increase in pregnancy rates (59%) compared with ewes subjected to sham lutectomy (33%; P < 0.05). In Expt 2, progestogen supplementation was initiated in the morning of day 4 after mating and continued until pregnancy diagnosis on day 25. In the afternoon of day 4, one ovary selected at random was lutectomized in ewes (n = 34) with at least one corpus luteum on each ovary, and the uterine horns were isolated by ligation to impede intraluminal transfer of luteal or uterine products that might initiate embryonic death. On days 5-8 after mating, equal numbers of ewes were injected i.m. with either saline or 5 mg PGF2alpha at 8 h intervals. Pregnancy rates did not differ between isolated uterine horns contralateral and ipsilateral to the regressing corpus luteum; however, pregnancy rates were lower in PGF2alpha-treated ewes than in saline-treated ewes (34 and 77%; P < 0.05). In conclusion, regressing corpora lutea exert an embryotoxic effect; however, there is no evidence that this effect occurs through systemic pathways.

During early pregnancy, pig conceptuses initiate the synthesis of oestrogens and on day 12 their trophoblastic membranes undergo a rapid expansion throughout the uterine horns. The insulin-like growth factor (IGF) system may be involved with conceptus development and steroidogenesis in pigs. Changes in uterine luminal IGF, insulin-like growth factor binding proteins (IGFBPs) and enzymatic activity for cleavage of IGFBPs during the oestrous cycle and early pregnancy were investigated. Uterine luminal content of IGF-I and IGF-II in uterine flushings from pigs on day 12 of pregnancy were two and three times greater, respectively, compared with uterine flushings collected from gilts during the oestrous cycle. Both IGF-I and -II content decreased on day 15 of gestation but content of IGF-II in uterine flushings remained three times greater than that of cyclic gilts. IGFBP-2 and -3 were the predominant binding proteins present in uterine flushings during days 0-10 of the oestrous cycle or day 10 of pregnancy. No IGFBPs were detected in the uterine flushings of either cyclic or pregnant pigs after day 10 by ligand blotting. Incubation of [125I]-labelled IGFBPs with various protease inhibitors indicated that cleavage of [125I]-labelled IGFBP-2 and -3 in uterine flushings involved serine proteases such as tissue kallikrein and matrix metalloproteinases. The results of the present study indicate that an increase in tissue kallikrein activity on day 12 of the oestrous cycle and pregnancy in pigs can directly, or indirectly through activation of matrix metalloproteinases, cleave IGFBP-2 and -3, thus allowing uterine release of IGF-I and -II in the uterine lumen to stimulate conceptus development.

During implantation, matrix metalloproteinases are believed to play roles in the tissue remodelling that accompanies decidualization in the endometrium and in embryo invasion. The objective of this study was to characterize further the expression of matrix metalloproteinases 2 and 9 in the mouse uterus during early pregnancy and oil-induced decidualization. mRNA encoding matrix metalloproteinase 2 was detected in pregnant uteri and uteri undergoing oil-induced decidualization by northern blot analyses. The steady-state concentrations of mRNA encoding matrix metalloproteinase 2 did not change significantly in implantation compared with inter-implantation areas on days 5-8 of pregnancy but were significantly lower in stimulated compared with non-stimulated uterine horns during artificially induced decidualization. mRNA encoding matrix metalloproteinase 9 was also detected in uteri undergoing oil-induced decidualization but not in pregnant uteri. Its concentration was significantly greater in uterine horns undergoing oil-induced decidualization compared with control horns. Immunoreactive matrix metalloproteinases 2 and 9 were detected in the uterus during early pregnancy and oil-induced decidualization by immunohistochemistry, localized to the endometrial stroma, but the staining progressively became weaker and was absent in areas that had undergone decidualization. By day 8 of pregnancy and 72 h after the induction of decidualization, matrix metalloproteinase 2 and 9 proteins remained mainly in the region of non-decidualized stromal cells adjacent to the myometrium. In implantation segments, they were also localized to the region of the trophoblast giant cells. The second objective of the present study was to determine whether endometrial stromal cells isolated from uteri sensitized for decidualization express matrix metalloproteinases 2 and 9. Northern blot analyses and gelatin zymography showed that these cultured cells expressed matrix metalloproteinase 2 and 9, and that transforming growth factor beta1 significantly increased matrix metalloproteinase 9 expression. The results of the present study further characterize matrix metalloproteinases 2 and 9 expression in the uterus during implantation and artificially induced decidualization.

Mouse embryo implantation depends on the complex interaction between the embryo trophoblast cells and the uterine environment, which deposits an extracellular matrix with abundant amounts of laminin. Intrauterine injection and blastocyst or ectoplacental cone culture models were used to study the effect of 32/67 kDa laminin-binding protein antibody on mouse embryo implantation in vivo and in vitro. Intrauterine injection of 32/67 kDa laminin-binding protein antibody (0.4 mg in 1 ml Ham's F-10 medium, 5 microl per mouse) into the left uterine horns of mice (n = 22) on day 3 of pregnancy inhibited embryo implantation significantly (P < 0.001) compared with the contralateral horns that had been injected with normal rabbit IgG. A continuous section study on day 5 after injection showed that the embryos in the control uteri implanted normally and developed healthily, but there were no embryos or the remaining embryos had disintegrated in the uteri injected with 32/67 kDa laminin-binding protein antibody. Blastocysts or ectoplacental cones were cultured in media containing 32/67 kDa laminin-binding protein antibody (0.2 mg ml(-1)) on laminin-coated dishes with normal rabbit IgG at the same concentration as in the controls. The 32/67 kDa laminin-binding protein had no effect on blastocyst or ectoplacental cone attachment, but prohibited the blastocyst or ectoplacental cone outgrowth and primary or secondary trophoblast giant cell migration. These results indicate that 32/67 kDa laminin-binding protein antibody blocked mouse embryo implantation by preventing embryo trophoblast cell invasion and migration through the uterine decidual basement membrane-like extracellular matrix which has a high laminin content.

Mares were inseminated with motile spermatozoa suspended in 30-150 microliters Tyrode's medium directly onto the uterotubal papilla at the anterior tip of the uterine horn, ipsilateral to the ovary containing a dominant preovulatory follicle of > or = 35 mm in diameter, by means of a fine gamete intrafallopian transfer (GIFT) catheter passed through the working channel of a strobed light videoendoscope. Insemination of 10, 8, 25, 14, 11 and 10 mares with, respectively, 10.0, 5.0, 1.0, 0.5, 0.1 or 0.001 x 10(6) motile spermatozoa resulted in conception rates of, respectively, 60, 75, 64, 29, 22 and 10%. Deposition of 1.0 x 10(6) motile spermatozoa onto the uterotubal papilla began to approach the limit of successful fertilization. These doses are far lower than the 3-15 x 10(9) spermatozoa normally ejaculated by fertile stallions during mating, and the accepted minimum dose of 500 x 10(6) spermatozoa used for conventional uterine body insemination in mares. The simplicity of the technique offers a practical means of exploiting new breeding technologies that require very small numbers of spermatozoa in horse breeding.

In rats, dams that conceive in their postpartum oestrus and then lose their firstborn litter bias the sex ratio of the litter toward females in utero. The present study identifies the source of litter sex ratio bias in these postpartum pregnant non-lactating dams. The female bias arises first through the postconception loss of embryos, and second, the loss occurs in midpregnancy between the attachment of the blastocyst to the uterine wall on day 5 and full metrial gland development on day 14. Some pregnancies were restricted to one uterine horn to see if this loss (and thus the opportunity for litter sex ratio biasing) was influenced by local factors operating within the uterine horn. Embryonic loss was more closely associated with the number of embryos implanting in a single horn than with the number implanting in the litter, demonstrating that local crowding within a horn is sufficient for the preferential loss of male embryos. This loss did not cause an obvious decrease in the size of the live-born litter because only those horns with a surfeit of embryos lost them. This process was the same in the right and left horns; both carried and lost the same numbers of embryos. A dam that conceives in her postpartum oestrus and then loses her suckling litter forgoes the implantation delay and uterine healing caused by lactation. Male embryos are less successful at implanting in a uterus only recently vacated by a previous litter.

Nitric oxide (NO) plays an important role in cell signalling in many physiological systems, including reproduction. During pregnancy, oestrogen modulates uterine NO generation, and NO may play an intermediary role in the oestrogen-mediated effects on the uterus. Since oestrogen is actively involved in inducing endometrial receptivity to support the process of implantation, the role of NO in the process of implantation in rats was investigated. N\g=w\-nitro-l-argininemethyl ester (l-NAME), an inhibitor of NO synthase (NOS), was administered in utero with or without sodium nitroprusside (SNP), a generator of NO, on different days during the preimplantation phase of gestation. The status of gestation in respect of implantation failure, endometrial receptivity and embryo development were assessed. l-NAME was administered at various doses (2\p=n-\5mg per uterine horn) and on different days of pregnancy (days 2\p=n-\6of pregnancy) to optimize the pregnancy terminating dose (absence of implantation site on day 8 of pregnancy) and the effective day of treatment. l-NAME led to failure of implantation when administered at 2.5 mg per uterine horn on day 3 of pregnancy. The characteristic preimplantation permeability changes in the uterus were significantly attenuated and embryo growth was retarded. The l-NAME-mediated effects were significantly reversed when SNP (100 \g=m\g)was co-administered with l-NAME. These findings suggest a role for NO in the process of implantation. The possible mechanism by which inhibition of the NO\p=n-\NOS system may interfere with implantation is discussed.

During early pregnancy, progesterone stimulates the secretion of proteins and other molecules that support the developing conceptus. Some gilts are able to support conceptus development as early as 110 days of age. The objective of this study was to evaluate the onset of responsiveness of the prepubertal uterus to progesterone. Thirty gilts were assigned to receive 2.2 mg progesterone kg-1 body mass per day or corn oil daily for 14 days starting at 6, 46, 76, 106, and 136 days of age. Hysterectomies were performed the day after the last treatment of progesterone, and the uterine horns were weighed and flushed with sterile saline (0.85% NaCl). Recovered flushings were analysed for total luminal protein, retinol binding protein, uteroferrin, prostaglandin E and prostaglandin F. An interaction between age and progesterone occurred for uterine wet mass (P < 0.001). Progesterone did not affect the uterine mass of gilts that underwent hysterectomy at 20 days of age, but did increase the uterine mass (P < 0.05) in other age groups. Progesterone increased (P < 0.01) the amount of total luminal protein in all but the youngest gilts. An increase in the amounts of retinol binding protein and uteroferrin (P < 0.001) by progesterone was first observed in 90-day-old gilts. Prostaglandins exhibited a different age-related pattern. The amount of prostaglandin E was increased (P < 0.001) by progesterone treatment in gilts aged 90-150 days, with a greater (P < 0.05) response at 120 days than at 90 days old. The response at 150 days old decreased (P < 0.05) to that observed at day 90. The response of prostaglandin F to progesterone followed a similar age-related pattern. Therefore, uterine responsiveness to progesterone develops between 20 and 90 days after birth, and uterine mass responds earlier than the secretory responses measured in our study.

The influence of a transcervical infusion of seminal plasma on preovulatory LH profiles and the advancement of ovulation after seminal plasma infusion for different times during oestrus were investigated using the single uterine horn infusion technique (Mariensee model), in combination with transcutaneous sonographic monitoring of the ovaries. Preparative surgery in 23 German Landrace gilts comprised the detachment of the left uterine horn from the corpus, leaving the caudal end open to the peritoneal cavity but sealing the corpus wound. In six gilts fitted with a permanent jugular vein catheter the patent horns were administered a transcervical infusion of seminal plasma (n = 5 cycles) or PBS (n = 4 cycles) immediately after the detection of oestrus by a teaser boar. In addition, 17 non-catheterized gilts received infusions of seminal plasma either 0 h (n = 3 gilts), 16 h (n = 7 gilts) or 24 h (n = 7 gilts) after the detection of oestrus. Seminal plasma infusion at the onset of oestrus provoked ovulation in the ipsilateral ovary of the treated horn 8.5 \m=+-\0.9 h earlier than in the contralateral (control) ovary. Seminal plasma did not influence the LH profile compared with PBS (P > 0.05), but shortened the interval between the LH peak and ipsilateral ovulation to 23.4 \m=+-\4.0 h compared with 31.8 \m=+-\3.4 h in the contralateral ovulation (P \m=le\0.01). Infusion 16 h after the onset of oestrus reduced the effect to 4.6 \m=+-\3.8 h with a wide range of 0\p=n-\8h (P < 0.01). The effect was more pronounced in gilts with long intervals between the onset of oestrus and contralateral ovulation compared with earlier ovulation on the control ovary. Seminal plasma infusion less than 16 h before contralateral ovulation and 24 h after the detection of oestrus had no effect. It is concluded that transcervical infusion of seminal plasma early in oestrus synchronizes the variable intervals between the onset of oestrus and ovulation in sows by a locally active mechanism.

We examined the key events underlying maternal recognition of pregnancy and the prevention of luteolysis in early postpartum ewes by synchronously transferring single expanded blastocysts recovered from control ewes on day 11 of pregnancy into the uterus of either postpartum recipients that had been induced to ovulate 28 days after lambing (n=12) or control recipients (n=11). Conceptus development, uterine flush interferon (oTP-1) concentrations, endometrial oxytocin receptor concentrations and endometrial prostaglandin F 2α (PGF 2α ) release in vitro were determined 5 days later (corresponding to day 16 of the ovarian cycle). By this stage, both conceptus mass and oTP-1 content of total uterine flush in the eight postpartum recipients that remained pregnant were significantly lower (P

The ovarian-uterine vasculature was examined in three Australian species of flying fox (Pteropus scapulatus, P. poliocephalus and P. alecto). Vascular casts and histological sections were used to determine the relationship between the blood supply and the localized endometrial reaction, which occurs ipsilateral to the ovulating ovary. The ovarian artery coils extensively just cranial to the ovary, gives off a branch to the ovary and continues caudally as the major vessel supplying the cranial tip of the uterus, where it anastomoses with the smaller uterine artery. The coil of the ovarian artery is completely enclosed by a venous sinus that drains the cranial pole of the ovary. The ovary is heavily encapsulated, with primordial follicles restricted to the caudal pole; thus, the corpus luteum is completely internal and placed cranially, close to the coil of the ovarian artery. This arrangement would allow countercurrent or crosscurrent transfer of ovarian steroids from ovarian vein to ovarian artery and on to the cranial tip of the ipsilateral uterine horn. The steroids could thus reach high concentrations locally and generate localized endometrial growth. Cranial to the coil, the ovarian artery is enclosed in a venous sinus that derives from uterine as well as ovarian veins. This would allow countercurrent transfer of bioactive substances from uterus to ovary.

In sheep, the presence of an embryo in utero on the 12th to 13th day after oestrus prevents luteolysis. These studies investigated whether platelet-activating factor (PAF) could exert an antiluteolytic function, either alone or in combination with interferon. The intrauterine administration of 250 micrograms PAF per horn per day, administered through indwelling cannulae into the uterus as injections twice a day (n = 12) or by continuous infusion (n = 4) failed to extend luteal function compared with controls (n = 8). When indwelling cannulae were used to administer (i) 125 micrograms PAF per uterine horn, as a bolus infusion twice a day (n = 5), (ii) continuous infusion of 500 micrograms bovine recombinant alpha 1-interferon each day (brIFN, n = 5), (iii) 125 micrograms PAF per horn twice a day, plus 500 micrograms brIFN per day (n = 8), or (iv) vehicle (n = 5), the luteal phase was significantly longer in co-infused (iii) than in control (iv) animals. These findings indicate that pharmacological doses of PAF may act synergistically with interferons to prevent luteolysis.

The effect of RU486, a synthetic progesterone receptor antagonist, on basal uterine prostaglandin (PG) release and release in response to oxytocin injection has been investigated in late-pregnant sheep (days 135–140 of gestation). Fifteen hours after i.m. injection of RU486 (50 mg; n = 5) or vehicle alone (n = 4), bolus injections of oxytocin (50, 500 and 5000 mU) were administered via a uterine artery ipsilateral to the pregnant uterine horn at 2-hourly intervals. Uteroovarian vein concentrations of 13,14-dihydro-15-keto PGF2α (PGFM) and PGE2 were determined before and during oxytocin stimulation. Basal concentrations of both PGFM and PGE2 were significantly (P < 0·001) increased in ewes 15 h after RU486 administration compared with ewes receiving vehicle alone. Concentrations of PGFM, but not PGE2, increased significantly (P < 0·001) following injection of each dose of oxytocin in both treated and untreated animals. The response to oxytocin, measured both as the area under the curve and as the peak height of PGFM release, was significantly (P 2 response in either RU486-treated or control animals. These results demonstrate that treatment of late-pregnant ewes with RU486 results in an increase in basal uterine PGFM and PGE2 as well as oxytocin-stimulated PGFM release. Journal of Endocrinology (1992) 134, 353–360

In Expt 1, activity of 2',5'-oligoadenylate (2',5'-A) synthetase in endometrium collected on Day 16 (oestrus is Day 0) from the uterine horn ipsilateral to the corpus luteum was greater (P less than 0.001) for pregnant (135.5 +/- 1.72 nmol/mg protein/h) than for cyclic ewes (58.5 +/- 0.99 nmol/mg protein/h). In pregnant ewes, activity of 2',5'-A synthetase in endometrium collected from the contralateral uterine horn (119.5 +/- 1.72 nmol/mg protein/h) did not differ from that of the ipsilateral horn. In Expt 2, three ovariectomized ewes were treated with progesterone for 10 days and then with oestrogen for 2 days. Activity of 2',5'-A synthetase on Day 13 was 18% greater (P less than 0.10) in endometrium collected from the uterine horn receiving infusions of 30 micrograms ovine trophoblast protein-1 (oTP-1) twice a day on Days 10, 11 and 12(57.7 +/- 0.22 nmol/mg protein/h) than from the uterine horn receiving control infusions of serum protein (SP; 48.8 +/- 0.22 nmol/mg protein/h). In Expt 3, activity of 2',5'-A synthetase on Day 15 was not significantly greater in endometrium collected from the uterine horn of cyclic ewes receiving infusions of 30 micrograms oTP-1 twice a day on Days 12, 13 and 14 (46.5 +/- 0.37 nmol/mg protein/h) than in endometrium from the uterine horn receiving infusions of SP (38.2 +/- 0.37 nmol/mg protein/h). When results of Expt 2 and Expt 3 were combined, intrauterine infusion of oTP-1 increased (P less than 0.05) activity of 2',5'-A synthetase in endometrium by 20%.(ABSTRACT TRUNCATED AT 250 WORDS)

Marked disparity in the uterine horn dimensions and relative degrees of caruncle development in suni suggested that exclusive or predominant dextral implantation occurs in association with bilateral ovulatory activity. Daily urinary measurements of pregnanediol-3 alpha-glucuronide revealed an oestrous cycle of approximately 21 days in length. Ovarian activity was controlled for synchronization of oestrus by using progestagen-impregnated intravaginal sponges and multiple ovulations were induced by using exogenous gonadotrophin therapy. An effective transcervical uterine catheterization technique was developed for the non-surgical collection of embryos. The efficiency of embryo recovery performed 5 days after sponge removal was 50.0%.

Follicular oocytes collected from prepubertal gilts at a local slaughter house were matured (36 h), fertilized and developed in vitro. Of 785 embryos, 190 (24%) embryos cleaved to the 2-4 cell stages with blastomeres of regular size by 33 h after insemination. These cleaved embryos were surgically transferred into the oviducts of 4 synchronized recipient gilts and recovered from the uterine horns 4 or 7 days later: 13 morulae, 2 blastocysts and 1 expanded blastocyst were recovered after 4 days and 3 hatched blastocysts were recovered 7 days after transfer. Re-culture in vitro sustained further development of morulae recovered 4 days after transfer: 11 of 13 morulae had developed to the blastocyst/hatched blastocyst stages. Overall, 17 of 190 (9%) embryos developed to the blastocyst stage. The results indicate that pig oocytes can be matured and fertilized in vitro, and subsequently develop to the blastocyst stage.

The gene for estrogen receptor a (ERa) has been shown to be under complex hormonal control and its activity can be regulated by mRNA alternative splicing. Here we examined the regulation of ERa transcription and translation in the rat uterus by ovarian steroid hormones. We examined whether expression of ERa mRNA splic isoforms is hormonally regulated in ovariectomized (OVX) and cycling rats. Adult OVX female rats were treated daily with 17-estradiol (E2) (0·05 μg/rat or 5 μg/rat), progesterone (P4) (1 mg/rat) or a combination of both hormones for 4 days. Animals were killed 24 h after the last injection and uterine horns were removed. In order to determine whether ERa mRNA isoforms are differentially expressed under various physiological conditions, animals were evaluated at proestrus, estrus and diestrus. The ERa protein and mRNA were detected by immunohistochemistry and comparative RT-PCR analysis respectively. The presence of ERa mRNA isoforms was evaluated using a nested RT-PCR assay. In OVX control rats, ERa mRNA and protein levels were high, demonstrating a constitutive expression of the ERa gene in the uterus. When animals received P4 or the high dose of E2,a significant decrease in both ERa mRNA and protein was observed in the uterus. However, when rats were treated with the low dose of E2, only the ERa protein was down-regulated; no changes were observed in ERa mRNA expression. In addition to the full-length ERa mRNA, OVX control rat uteri expressed three shorter transcripts: a3, a4 and a3,4 (lacking exon 3, exon 4, or both 3 and 4 respectively). Surprisingly, when OVX animals were treated with P4, the low dose of E2 or a combination of both steroids, expression of the a3 isoform was completely abolished. During the estrous cycle, all ERa mRNA splicing variants were detected at proestrus and estrus. However, in diestrus, significant low levels of the a3 isoform were observed. In summary, our results suggest a dose-dependent relationship between E2 concentrations and the level of control in the ERa transcription–translation cascade. Moreover, the alternative splicing of the ERa imary transcript is influenced by the hormonal milieu, suggesting that these events could affect the estrogen responsiveness of the rat uterus during the estrous cycle. Fil: Varayoud Jorgelina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina Fil: Ramos J Guillermo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina Fil: Monje, Lucas Daniel. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina Fil: Bosquiazzo, Veronica Lis. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina Fil: Muñoz de Toro, Monica Milagros. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina Fil: Luque, Enrique Hugo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina

The present study was undertaken to determine whether the mechanism of embryo transfer is a factor in the action of epidermal growth factor (EGF) in initiation of implantation. Unilateral intrauterine infusion of 3 microliters buffered saline, or saline containing 1.5 micrograms EGF, plus i.v. injection of 100 micrograms EGF 2 h later resulted in implantation sites in all animals within 48 h. In several animals implantation was also initiated in the non-injected uterine horn. Administration of indomethacin 1 h before the intrauterine injection completely blocked the effect of EGF but not that of 25 ng oestradiol. The results confirm that EGF can replace oestrogen for initiation of implantation provided that the uterine trauma associated with embryo transfer, that is puncture, is provided. The mechanisms involved remain to be resolved.

Classical pharmacological studies have shown that oestrogen dominance in humans and other animals can increase the responsiveness of the uterus to many locally acting peptides. Parathyroid hormone-related protein (PTHrP) has been shown to be expressed in the pregnant and non-pregnant rat uterus and exogenous PTHrP is known to relax uterine contraction in vitro. We investigated whether oestrogen dominance can influence the responsiveness of the uterine horn to PTHrP, and further studied the localization of PTHrP mRNA and protein in the rat uterine horn using in-situ hybridization and immunohistochemistry. Exogenous PTHrP(1–34) inhibited spontaneous and electrically induced contractions in uteri isolated from non-cycling rats. Pretreatment of non-cycling rats with oestradiol-17β increased uterine sensitivity to PTHrP: EC50 values for inhibition of spontaneous contractions by PTHrP were 0·33 nmol/l, 1·1 nmol/l, 2·6 nmol/l and 7800 nmol/l in uteri from animals treated for 2 days with oestradiol-17β alone, 2 days with oestradiol-17β+1 day progesterone, 1 day with oestradiol-17β alone and in untreated rats respectively. Similar EC50 values were obtained for electrically stimulated uteri. In agreement with these findings, uterine horns from cycling rats in pro-oestrous and oestrous phases of the cycle showed a higher responsiveness to PTHrP(1–34) when compared with uterine horns taken from rats in metoestrus and dioestrus. PTHrP mRNA and protein were detected in the endometrial epithelium lining of the lumen and the endometrial glands, as well as in the myometrium of rats which were either pretreated for 2 days with oestradiol-17β or untreated. This study suggests that PTHrP may act in an autocrine and/or paracrine manner to modulate uterine motility and function. Journal of Endocrinology (1992) 134, 415–425

There has been little success in inducing a decidual reaction in the uteri of adult female rats given androgen neonatally. Burin, Thevenot-Duluc & Mayer (1963) reported a positive reaction after scratching the antimesometrial lining and giving progesterone (5 mg) daily for 8 days, but the incidence of success, the age of the animals, and the criteria for decidualization were not given. Barraclough (1967) reported that 25% of adult females given HCG to induce ovulation, and reserpine to sustain pseudopregnancy, produced a positive decidual reaction. All other reports are negative. The reasons for failure are not clear. Lack of oestrogen-binding capacity has been documented (McGuire & Lisk, 1969) and may play an important r\l=o^\le;the availability of progesterone receptors in the uteri of such females is unknown. In the present study, attempts were made to induce a decidual cell reaction in the uteri of adult females of various ages given different doses of testosterone propionate (TP) at 5 days of age. Holtzman strain female rats were given 50 \g=m\g, 250 \g=m\g or 1250 \g=m\g TP on the 5th day after parturition. A decidual reaction was induced in adult females by scratching the entire antimesometrial length of one uterine horn; the contralateral horn served as the control. A 300% increase in weight of the former over the latter was considered significant (DeFeo, 1963). Progesterone was administered subcutaneously dissolved in 0\m=.\1 ml sesame oil, as was oestradiol benzoate (OB) when it was used. Animals were killed on the 10th day after treatment began. Young adults were given progesterone for 9 days and were killed on the 10th day (Table 1 ). Uterine trauma on Day 6 elicited a response in all rats (Groups 2A and 3A). The need for progesterone was indicated by the lack of response with oil or with early withdrawal of treatment. The 2 mg dose of progesterone was used in all further tests. The progesterone apparently caused a reduction in gonadotrophin from the pituitary because ovarian weight was reduced (Groups 2A and 3A versus 4A and 5A). A positive response was obtained in about 25% of 3to 4-month-old females given 1250 μg TP neonatally (Table 1). Again, oestrogen did not increase the incidence of positive responses. The steroid treatment reduced the ovarian weight of the rats in three of the four groups. These results are similar to those

Embryos were collected from the oviducts and uterine horns of mice bearing a unilateral IUD and of non-IUD control mice at various times p.c., examined for the presence or absence of polymorphonuclear leukocytes (PMNLs) and cultured. Normal numbers of embryos were obtained on the IUD side at 48 h p.c., but reduced numbers were recovered at 60-66 and 72 h p.c. The numbers of embryos that hatched from those collected from control, IUD-bearing and contralateral non-IUD bearing tracts were, respectively, 81, 29 and 36% for 48-h embryos, 78, 47 and 51% for 60-66-h embryos, and 88, 47 and 54% for 72-h embryos. At 48 h and 60-66 h p.c. the arrest of development occurred at the expanded blastocyst stage, but at 72 h p.c. some compacted morulae were affected. Some embryos associated with PMNLs at collection developed normally when the PMNLs were dispersed before culture. An initial association with leukocytes is therefore not necessarily detrimental to the subsequent in-vitro development of the embryo in mice.

Day 7 eggs (oestrus = Day 0) were surgically transferred singly to the uterine horn contralateral to the CL and heifers were either slaughtered on Day 24-26 (Group 1) or palpated per rectum on Day 42 (Group 2) to determine the presence of a developing conceptus and a maintained corpus luteum. There was a fall in pregnancy rate of the control heifers in Groups 1a (6/10) and 2a (2/10) (P = 0.085). Pregnancy rate was unaffected by progesterone treatments from Day 13 to the day before slaughter in Group 1b (8/10), or to Day 23 in Group 2b (2/10). Two of the conceptuses recovered in Group 1b were developing in the presence of regressed corpora lutea. There was, however, an increase in pregnancy rate in Group 2d (6/10) compared with Group 2a (2/10) when heifers were treated with hCG from Days 13 to 35 (P = 0.085). The results indicated that embryonic death is occurring after Day 24 and suggest that a change in the nature of the embryonic signal may be occurring at this time.

In Exp. 1, administration of 5 mg oestradiol valerate i.m. to pregnant gilts on Days 9 or 9 and 10 advanced the uterine secretion of calcium, protein, and acid phosphatase as demonstrated by levels recovered in the uterine flushings of females unilaterally hysterectomized on Day 11. Upon removal of the remaining uterine horn on Day 12, protein and acid phosphatase increased while Ca2+ decreased in oestradiol-treated gilts as did PGF. In contrast, a 4-fold increase in recoverable Ca2+ occurred from Days 11 to 12 in control gilts. Recoverable oestradiol-17 beta was increased in all 3 groups on Day 12 and plasmin inhibitor concentration increased in oestradiol-treated gilts. Two-dimensional PAGE demonstrated the appearance of a group of very acidic polypeptides in oestradiol-treated gilts. Blastocysts recovered from the second uterine horn had undergone elongation to the filamentous morphology in all 3 groups. In Exp. 2, oestradiol valerate was administered to pregnant gilts on Day 9 or Days 9 and 10 followed by total hysterectomy on Day 16. No differences in recoverable Ca2+ or protein were found, but acid phosphatase was decreased by 75% after oestradiol treatment. Recoverable oestradiol was decreased in oestradiol-treated gilts while PGF and plasmin inhibitor concentrations were unaffected. Compared with the control gilts, blastocysts recovered from oestradiol-treated gilts were fragmented and degenerating on Day 16. PAGE demonstrated greatly intensified staining of the group of acidic polypeptides in oestradiol-treated gilts. These results indicate that oestradiol treatment on Day 9 of pregnancy advances uterine secretory response, but that blastocyst elongation can occur in this uterine environment and in the presence of declining intraluminal Ca2+ levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Inequality in function of the left and right ovaries and uterine horns of mice was evaluated in three separate experiments. In Exp. 1, the effect of position in the reproductive tract on various reproductive characteristics was evaluated in 158 pregnant hybrid mice. Ovulation rate, number of fetuses, total fetal weight and total placental weight were higher (P less than 0.05) on the right than the left on Day 18 of pregnancy (vaginal plug = Day 1). In Exp. 2, the effect of previous sham or unilateral ovariectomy (right or left) in mated Swiss-Webster mice was compared with unoperated mated controls (N = 17-24/treatment). In control mice, ovulation rate, total fetal weight and ovarian weight were higher (P less than 0.05) on the right than left side. Surgery (sham or unilateral, ovariectomy) decreased (P less than 0.005) ovulation rates, number of fetuses, ovarian weights, total fetal weight and total placental weight on Day 18 of pregnancy. Unilateral ovariectomy decreased (P less than 0.05) ovulation rates and ovarian weights more than did sham operation. Ovulation rates were higher (P less than 0.01) when the left ovary was manipulated or removed rather than the right ovary. For Exp. 3, pairs of 8 hybrid mouse embryos each (morulae and blastocysts) were surgically transferred to the left and right uterine horns of the same (bilateral, N = 15) or different (unilateral, N = 28) Swiss-Webster recipients. In almost all incidences, embryo survival (to Day 18 of pregnancy) was twice as high (P less than 0.05) in right than left uterine horns. We conclude that the left and right ovaries and uterine horns are not equal in function in Swiss-Webster and a hybrid strain of mice.

A study to determine whether aspirin or indomethacin inhibit uterine horn hypertrophy in rats fitted with a silk suture IUD is presented. Nonovariectomized animals received per kg body weight either 1) hydrocortisone 12.5 mg 2) indomethacin 2 mg 3) aspirin 10 mg 4) aspirin 100 mg or 5) meclafenamate 2 mg. Ovariectomized animals received 2 mg/kg body weight of indomethacin. The lower dose of aspirin and hydrocortisone failed to inhibit a weight increase in the uterine horn of nonovariectomized rats. The larger dose of aspirin indomethacin and meclafenamate partily but significantly inhibited a weight increase (p less than .001). In the ovariectomized rats the IUD caused an increase in the uterine wet weight compared with the contralateral control horn. The administration of indomethacin partly but significantly reduced the uterine horn weight gain in the ovariectomized animals (p less than .005). The increased weight may be due in part to a continuous release of prostaglandins stimulated by the presence of an IUD.

The highest values of gamma-aminobutyric acid (GABA) in the genital tract of the rat at different stages of the oestrous cycle were found in the oviduct (3.5-7 micrograms/mg protein) and the lowest in the ovary (50-100 ng/mg protein). The values for uterus and vagina ranged between 80 and 150 ng/mg protein. GABA (10-30 ng/microliter) was also found in fluid in the ovarian bursa. At 11:00 h, on the day of oestrus, GABA content increased in the ovaries but values in the oviducts were maximal at 11:00 h on the day of pro-oestrus. Variations in GABA content of the vagina were also found. Uterine cervix or uterine horn showed no changes during the oestrous cycle. The GABA content was not uniform throughout the oviduct: the highest values were found in the portion next to the ovary. At 10 days after removal of the right oviduct, GABA values in the ovary and ovarian bursa fluid decreased on the operated side. At 1 month after surgery, the values in ovary were normal but the values in ovarian bursa fluid were still low, suggesting that the source of ovarian GABA was not the oviduct. The variations observed in the present paper suggest an involvement of GABA in reproductive physiology.

Endometrium was obtained from cattle slaughtered at various stages of early pregnancy and of the oestrous cycle. Analyses for protein, RNA, DNA, glucose and some lysosomal enzymes were carried out on this tissue. The results are considered with respect to the general influence of the hormonal status of pregnancy and the specific influence of the hormonal status of pregnancy and the specific influence of the blastocyst in one uterine horn.

The distribution of activity between the left and right sides of the reproductive tract, as measured by numbers of CL, embryos and placental scars, was studied in small mammals of 22 species. Shrews ovulate from the two ovaries in a distribution that does not differ from the binomial. Implantation of blastocysts in the two uterine horns is more nearly even ('balanced') than would be predicted from the binomial distribution. Balance in this group apparently is achieved by transuterine migration of blastocysts, perhaps in conjunction with some spacing mechanism within the uterus. Some cricetid rodents show little or no balance, but in others the distribution of activity sites (embryos, CL and placental scars) departs significantly from the binomial distribution. Reproductive activity sites of heteromyid and geomyid rodents (Geomyoidea) are highly balanced; uterine balance apparently is achieved by means of ovarian rather than uterine control. We know of no previous reports of ovarian balance and suggest that physiological mechanisms controlling numbers of ovulations in the species exhibiting this characteristic may differ from those in species exhibiting a random distribution of ovulation sites. Hypotheses regarding evolutionary aspects of balance are considered in phylogenetic and ecological terms, generating several testable research questions for physiologists, anatomists, and evolutionary ecologists.

Myoelectrical activity in the uterus of late-pregnant sheep was recorded by the chronic implantation of bipolar electrodes in the myometrium. Three distinct patterns of electrical activity were recognizable: irregular trains of action potentials (greater than 120 sec), long spike bursts (7--120 sec), or short spike bursts (less than 7 sec) each exhibiting high (greater than 100 muV) or low (50--100 muV) amplitude. The duration of spike activity was expressed as the myoelectrical index (MI), i.e. the fraction of the hour during which a given category of activity is exhibited. The increase in the overall MI which represents daily average values regardless of the class of amplitude from 72 h pre partum up to parturition was significantly (P less than 0.05) more pronounced in the uterine horn than in the body of the uterus. During the last 6--7 days of gestation high-amplitude irregular trains and long spike bursts were significantly (P less than 0.05) more frequent than these activity patterns with low amplitude. Regardless of the amplitude, the incidence of irregular trains began to decline rapidly 48 h pre partum with an abrupt cessation following parturition. The frequency of long spike bursts increased 72 h pre partum to reach a maximum which coincided with parturition.

Rats were thyroidectomized at different stages of gestation and killed at parturition. Acid phosphatase of uterine horns was significantly (P < 0.05) lower in animals thyroidectomized at later stages of pregnancy, whereas alkaline phosphatase was significantly lower in those thyroidectomized earlier as compared to intact pregnant animals. The activities of glutamic oxaloacetic acid and glutamic pyruvic acid transaminases also showed significant alterations in some of the thyroidectomized animals. These 4 enzymes were also studied in the cervix but the changes showed a less distinct pattern.

The reproductive tracts of 112 female little bulldog bats collected around the onset of a breeding season in the Cauca Valley of Colombia were examined histologically. Of the 88 females with luteinizing/luteinized follicles or new CL, 72 carried tubal ova or uterine blastocysts and 16 were non-pregnant. In 14 of the latter follicle rupture had apparently failed to occur, and the oocyte or a collapsed zona pellucida was found within a luteinizing or luteinized follicle. These structures appeared to be functional because most of the affected bats demonstrated preferential stimulation of the ipsilateral oviduct and/or uterine horn. Two of these animals also had a second, older luteinized follicle which contained the remnants of an oocyte. None of these 14 bats exhibited the uterine modifications thought to be associated with a previous pregnancy or had prominent mammary glands. Such reproductive features were, in contrast, frequently demonstrated by other females in the population. These observations suggest that the luteinization of unruptured follicles may have occurred in prepubertal members of the population and reflect immaturity of the hypothalamo-pituitary -ovarian axis in these individuals.

Uterine electromyography was performed by means of chronically implanted surface electrodes in 3 Pony mares during spontaneous oestrous cycles and following luteolysis induced by a prostaglandin analogue (fluprostenol). Three distinct patterns were recognized during the oestrous cycle. (1) During oestrus well defined phases of activity with closely grouped high-amplitude spikes were separated by long periods (10-45 min) of complete inactivity. (2) During dioestrus more diffuse phases of activity with low-amplitude spikes were separated by variable periods of relative inactivity. (3) During luteolysis, short and frequently occurring phases of activity were propagated between the two electrodes on one uterine horn; a similar pattern also occurred between 1 and 3 h after injection of fluprostenol. Peripheral plasma progesterone, but not total inconjugated oestrogen, concentrations were closely related to characteristics of the myographic activity during the cycle. Insemination during oestrus and injection of fluprostenol during dioestrus caused a marked and prolonged increase in myometrial electrical activity. Almost any non-specific environmental stimulus, including entry by palpation of the genital tract per rectum and vaginoscopic examination, but these were of brief duration and the normal resting pattern of activity was quickly re-established after completion of the manipulations.

The luminal epithelium of the mouse uterus was removed simply and rapidly by opening the uterine horns longitudinally and 'vortexing' them in buffer with glass balls. This procedure, which requires no specialized apparatus, was developed for use with uteri in different hormonal states, and 2 uterine horns/tube, 5 glass balls and vortexing for 2 min are suggested for routine use. Under these conditions, 79--100% of the cells were removed, yielding epithelial fractions of 65--90% purity. The suspension of cell constituents recovered contained intact nuclei which could be further purified. Extensive histological examination demonstrated that the other uterine tissues were minimally damaged.

An attempt has been made to assess quantitatively the extent of cell death in the uterine epithelium after oestrogen treatment. [3H]Thymidine was injected into ovariectomized mice at an interval after oestrogen treatment when many of the luminal epithelial cells were in the S phase of mitosis. Uptake of [3H]thymidine was confirmed by autoradiography of sections of uterus and scintillation counting of trichloracetic acid-insoluble fraction of whole uterine horns. Radioactivity declined after the cessation of oestrogen treatment but remained high if treatment was continued. The decline appears to be correlated with the cell death previously demonstrated in histological sections of uteri under similar conditions.

The numbers of lymphocytes occurring in 5 mm of uterine epithelium were counted in cyclic heifers on Days 5, 10, 15 and 20 after oestrus and in pregnant heifers between Days 19 and 27 after insemination. Specimens were taken from the oviducal, middle and body regions of the ipsilateral and contralateral uterine horns of the pregnant and cyclic animals. Mean +/- s.e.m. lymphocyte numbers in cyclic animals were 25.8 +/- 1.5 on Day 5, 19.7 +/- 5.5 on Day 10, 22.0 +/- 0.6 on Day 15 and 28.2 +/- 1.5 on Day 20 after oestrus. There was a significant reduction in lymphocyte numbers as the chorion and uterine epithelium developed into a functional placenta from 24.5 +/- 1.1 on Day 19 to 9.0 +/- 0.9 on Day 27. Qualitative observations suggested an increase in degenerating cells concomitant with the decrease in lymphocytes. Some of these degenerating cells may have been lymphocytes while others were probably uterine epithelial cells. As gestation progressed and the placenta was established, the number of potentially damaging cells at the fetal-maternal interface was reduced.

SUMMARY The release of prostaglandin-like material and the spontaneous contractions of individual horns from the pregnant rat uterus in vitro have been studied on day 22 of pregnancy – the expected day of delivery. Removal of foetuses (retaining placentae in utero) from one or both uterine horns on day 16 or 17 significantly reduced prostaglandin F release and spontaneous activity. Rats which had been made unilaterally pregnant after ligation of one uterine horn, exhibited a decrease in prostaglandin F output from both horns. Uterine activity and prostaglandin release were increased in quiescent uteri by the addition of arachidonic acid (5 μg/ml) or phospholipase A (160 mu./ml); these effects were abolished by indomethacin (20 μg/ml). However, the stimulation of uterine activity by PGF2α (30–60 ng/ml) was not affected by indomethacin. It is concluded that the release of prostaglandins from the pregnant rat uterus in vitro at term is related to the presence of viable foetuses.

Rats were unilaterally ovariectomized on Day 17 of pregnancy and the times of onset of parturition were recorded. Unilateral ovariectomy did not delay the onset of parturition in bilaterally pregnant rats. In rats which were unilaterally pregnant (spontaneously or after ligation of one uterine horn), removal of the ovary on the side of the gravid horn led to delayed or partial parturitions. When fetuses and placentas were removed from one uterine horn on Day 13 and the ovary from the opposite side on Day 17, or when the uterine artery and/or vein were ligated on one side and the ovary removed from the other, about half of the parturitions were abnormal. It is suggested that a substance produced by the gravid uterine horn is transmitted to the ovary, primarily by a local circulatory system, and thus initiates the onset of parturition.

Five domestic ferrets previously maintained for 12 weeks under a 16L:8D photoperiod were electroejaculated weekly for 15-65 weeks while continuing to be exposed to the prolonged light cycle. Two ferrets sustained spermatogenesis for 20 and 26 weeks, while sperm production in the remaining males either was sporadic or decreased, remained depressed and then increased to peak levels observed in other males. Regardless of the temporal spermatogenesis patterns within males, the number of electroejaculated spermatozoa with residual cytoplasmic droplets or abnormal acrosomes increased in all ferrets over time. Diluted ejaculates meeting artificial insemination criteria were deposited intravaginally or by transabdominal laparoscopy into the uterine horns of females treated 0 or 24 h earlier with 90 i.u. hCG. Vaginal insemination was ineffective (0 pregnancies in 10 attempts), but 17/24 ferrets (70.8%) inseminated laparoscopically became pregnant and delivered live young (mean litter size, 5.2 kits). Number of motile spermatozoa deposited in utero (1.6-10.0 x 10(6) cells), presence of glycerol in the sperm dilution medium (0 versus 4%) and time of hCG administration (0 versus 24 h before insemination) had no effect on pregnancy results or litter size.

This work was aimed at testing whether oestrogen-induced changes in the uterus are responsible for uterine eosinophil migration or whether the effect is due to a direct action of oestrogen on the eosinophils themselves. Uterine eosinophil migration in response to intraluminal administration of oestradiol-17β into one uterine horn was measured in the injected and in the untreated contralateral horns. Uterine genomic responses (luminal epithelial and myometrial hypertrophy and nucleolar enlargement), known to depend on the local intra-uterine oestrogen level, were measured to control for the absence of hormone recirculation to the uninjected horn. The effects of intravenously administered oestradiol were also determined. Intraluminal injection of 0·1 ng oestradiol had no effect on either horn. With 10 ng, only the injected horn exhibited the genomic responses while eosinophilia developed to the same extent in both horns. With 100 ng, the genomic responses and eosinophilia were identical in injected and in contralateral horns. The results show that eosinophil migration depends on systemic levels of oestrogen, thus indicating that it is due to an oestrogen-induced change in a property of eosinophil leukocytes rather than a change in the uterus itself. As water imbibition showed the same pattern of responses as eosinophilia, this lends further support to the hypothesis of a role for eosinophils in oestrogen-induced uterine oedema.

In Exp. 1, endometrium was collected from Day-15 cyclic ewes and effects of oTP-1, oxytocin and oTP-1 + oxytocin, in various temporal relationships, on phosphatidylinositol (PI) turnover were determined. Co-treatment of endometrium with oTP-1 and oxytocin inhibited stimulatory effects of oxytocin, while treatment with oTP-1 before and during oxytocin administration had no effect. Turnover of PI was unaffected by oTP-1 alone. In Exp. 2, ovariectomized ewes were treated with progesterone (50 mg/day) for 10 days and then oestrogen (100 micrograms/day) for 2 days and endometrium was collected. Oxytocin stimulated PI turnover in endometrium, but oTP-1 had no effect alone or in combination with oxytocin. In Exp. 3, ovariectomized ewes were treated with corn oil (1 ml/day), oestrogen (50 micrograms/day), progesterone (50 mg/day) or progesterone + oestrogen for 10 days and endometrium was collected. Oxytocin stimulated PI turnover only in ewes that received progesterone. oTP-1 alone had no effect on PI turnover, while co-treatment of endometrium with oxytocin and oTP-1 stimulated PI turnover in ewes treated with progesterone, but not progesterone and oestrogen. Pretreatment of endometrium with oTP-1 stimulated PI turnover when ewes were treated with progesterone or progesterone + oestrogen. Pretreatment of endometrium with oxytocin and then treatment with oTP-1 inhibited PI turnover compared to treatment with oxytocin alone. In Exp. 4, ovariectomized ewes were treated as in Exp. 2. Catheters were placed into the uterine horns and ewes received oTP-1 into one horn and serum into the other twice daily on Days 10-12 of steroid treatment. Endometrium collected on Day 13 was used to measure PI turnover and received either no treatment or oxytocin. Oxytocin stimulated PI turnover in endometrium of these ewes and in-vivo treatment of the ewes with oTP-1 had no effect on PI turnover. These results indicate that antiluteolytic effects of oTP-1 are not mediated by inhibiting effects of oxytocin on phosphatidylinositol turnover if oxytocin receptors are present and that uterine responsiveness to oxytocin is progesterone dependent.

The possible selection of spermatozoa for fertilization by the female genital tract was investigated using genetically homogeneous, numerically adjusted 'cervix-selected' and 'unselected' rabbit spermatozoa. Samples of spermatozoa were marked by 15 min exposure to 1.3 mg TEPA/ml and then washed for 15 min. TEPA-treated and control samples were inseminated alternately into the vagina (= 'cervix-selected') or uterine horns (= 'unselected') of prospective donors. After 6 h spermatozoa were recovered from the uterine horns of the donors. Equal numbers of 'selected' and 'unselected' spermatozoa were inseminated either into the uterine horns (24 does) or oviducts (25 does) of recipients. The fertilization rates were 48 and 72%, respectively. Significantly more eggs were fertilized by untreated than by TEPA-treated spermatozoa. Almost identical fertilization rates, however, were observed between 'cervix-selected' and 'unselected' spermatozoa. It is concluded, therefore, that in the rabbit no selection of (preincubated) spermatozoa for fertilization takes place at the cervical level.