Changes in the density of peripheral benzodiazepine binding sites in genital organs of the female rat during the oestrous cycle

The affinity and the density of peripheral-type benzodiazepine binding sites (PBzS) in tissues from the genital organs of female rats were studied during the oestrous cycle. When comparing PBzS density on the day of oestrus to PBzS density on the day of pro-oestrus, a significant increase was observed in the ovary (1.9-fold), oviduct (2.4-fold) and uterus (1.7-fold), but not in the kidney. Serum oestradiol also increased to a maximum on the day of pro-oestrus. The ovarian and uterine PBzS density and serum concentrations of oestradiol and progesterone were measured every 8 h between the days of dioestrus and pro-oestrus. Ovarian and uterine PBzS density increased to a maximal value at 01:00 and 09:00 h, respectively, on the day of pro-oestrus. However, a significant increase in PBzS density occurred in the ovary (P less than 0.02) and uterus (P less than 0.001) at 09:00 h on the day of pro-oestrus as compared to 09:00 h on the day of dioestrus. These changes were associated with an increase in serum oestradiol and progesterone concentrations. The affinity of PBzS in all tissues examined remained unaltered during the oestrous cycle. This study demonstrates that changes associated with the oestrous cycle occur in the density of PBzS in various genital organs.     

Immunoglobulins in the mouse uterus during the oestrous cycle

The distribution of IgA, IgG and IgM was studied by an immunoperoxidase technique on sections of mouse uteri at each stage of the oestrous cycle. Staining for IgG and IgA was highest at pro-oestrus, declined at oestrus and was very low during the other stages. At pro-oestrus IgG was found throughout the stroma, in the uterine lumen, and in 10% of glandular lumina; very few IgG-containing plasma cells were present. At pro-oestrus, IgA was found in the uterine lumen, and in most of the uterine glands, both in the lumen and in the epithelium; little IgA was present in the stroma. IgA-plasma cells were detected at each stage of the cycle and were particularly numerous at pro-oestrus and oestrus. These results suggest that IgA is secreted locally from plasma cells into the uterine gland through the glandular epithelium, but that IgG enters the stroma from the local capillaries. The obvious increase in IgG and IgA secretion at pro-oestrus, when plasma oestradiol levels are highest, supports the hypothesis that, during the oestrous cycle, the humoral immune response is regulated in the uterus by ovarian hormones.     

Changes in inhibin activity, and progesterone, oestrogen and androstenedione concentrations, in rat follicular fluid throughout the oestrous cycle

Follicular fluid was aspirated from all visible surface follicles of rats at selected times of the oestrous cycle. Fluids from a pair of rat ovaries were pooled and assayed for inhibin activity by the rat anterior pituitary cell culture assay. Serum LH, FSH and progesterone as well as follicular fluid progesterone, total oestrogens and androstenedione were also measured. Follicular fluid inhibin activity was relatively constant throughout the oestrous cycle (30.7 +/- 3.4% inhibition of FSH per 0.1 microliter follicular fluid) except for a well defined surge at pro-oestrus (09:00-16:00 h, peak at 14:00 h = 84.0 +/- 7.2% inhibition of FSH per 0.1 microliter follicular fluid). The follicular fluid was not treated with charcoal before assay because a pilot experiment showed that such treatment did not alter the inhibin activity of follicular fluid. Steroids in follicular fluid were generally lowest on the afternoon of oestrus and the morning of dioestrus I and generally elevated during pro-oestrus.     

Gamma-aminobutyric acid in the genital tract of the rat during the oestrous cycle

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.     

Attainment of puberty in female pigs: Clinical appearance and patterns of progesterone,oestradiol-17β and LH

The object of the study was to investigate the clinical and endocrine patterns of progesterone, oestradiol-17β and LH during the peripubertal period in female pigs. Crossbred gilts were penned in groups at an age of 10–12 weeks and boars were kept in adjacent pens during the entire experimental period. Daily oestrous checks started at 4.5 months of age and the gilts were slaughtered after their third heat. At the age of 4.5–5 months a permanent catheter was inserted in the cephalic vein and blood samples were collected from the gilts once daily until either the first or second oestrus. In three gilts hourly blood samples were taken during their first and second oestrus, beginning at early pro-oestrus.The gilts showed their first oestrus at the average age of 183 days. No corpora lutea from earlier ovulations were observed in gilts laparoscoped after their first detected oestrus. During the 30-day period before first oestrus the mean daily progesterone levels varied between 32 and 329 pmol/l. The average levels of oestradiol-17β varied between 15.6 and 30.8 pmol/l. There was no tendency for the oestradiol-17β level to rise before onset of first pro-oestrus. The average levels of LH varied between 0.15 and 0.94 μg/l. The statistical analyses revealed no significant relationship between the level of the hormones studied and onset of first oestrus. The mean progesterone levels during the first and second oestrous cycles were almost identical, however. Oestradiol-17β increased gradually during pro-oestrus, reaching maximum levels before onset of oestrus and thereafter decreasing sharply to values around 30 pmol/l. The oestradiol-17β levels were higher at the second than at the first pro-oestrous period. The concentrations of plasma LH rose sharply with declining plasma levels of oestradiol-17β. The duration of elevated plasma LH levels (> 1 μg/l) was, on average, 26 h and the LH levels were higher during the first oestrus than during the second oestrus. The first rise in progesterone was observed 11–29 h after the LH levels had decreased to concentrations below 1 μg/l.     

Expression and localisation of thioredoxin in mouse reproductive tissues during the oestrous cycle

Thioredoxin expression within the reproductive tissues of the female mouse was analysed during the oestrous cycle stages of dioestrus, oestrus and metoestrus by Western blot analyses and immunocytochemistry. From Western blot analyses the expression of thioredoxin was found to be increased in oestrus compared to dioestrus and metoestrus. Localisation of thioredoxin within the reproductive organs of the mouse during the oestrous cycle has shown that the expression of thioredoxin is specific for distinct areas within the reproductive organs. These areas are the stratified squamous epithelium of the vagina, the simple columnar epithelium and the uterine glands of the uterus, the ciliated columnar epithelium of the oviduct, the corpus lutea, the interstitial cells and the secondary follicles of the ovary. The discrete cellular localisation and oestrous dependence of thioredoxin expression are suggestive of specific roles in various reproductive processes.     

In-vivo microscopy of the rat endometrial subepithelial capillary plexus during the oestrous cycle and after ovariectomy

Blood flow through the endometrium was visualized by using incident-light fluorescence microscopy and a video image recorded for later detailed analysis. The subepithelial microvascular density was calculated for each day of the oestrous cycle and at 7 days after ovariectomy. The results showed that the microvasculature was significantly more dense at dioestrus I, pro-oestrus, and after ovariectomy than at oestrus, with dioestrus II being in between. Mean capillary path lengths running from arteriole to venule were longest at pro-oestrus, followed by oestrus, dioestrus II, dioestrus I, and shortest after ovariectomy. The results suggest that endometrial growth and regression precede microvascular growth and regression. The technique of in-vivo microscopy provides an important new avenue for investigating the role of local factors in the control of the endometrial microcirculation.     

Immunocytochemical localization of nitric oxide synthase-III in reproductive organs of female rats during the oestrous cycle

Summary Constitutive endothelial nitric oxide synthase (NOS III) expression during the oestrous cycle was mapped immunocytochemically on 5 μm-thick paraffin sections of rat female reproductive organs. Ovarian NOS III immunoreactivity increased with follicular maturation (strongest in dioestrus corpora lutea), suggesting that nitric oxide may regulate folliculogenesis and luteal functions. Oviductal NOS III, localized in mucosal epithelium and muscular wall, was maximal during pro-oestrus and oestrus, suggesting that nitric oxide may impart periovulatory quiescence for reception, retention and fertilization of ovulated oocytes. Uterine NOS III, localized in endometrial and glandular epithelium, and in myometrial smooth muscle cells, was abundantly expressed during pro-oestrus and oestrus. The peri-implantation period in pregnant rats corresponds to the periovulatory period and the elevated NOS, and thus nitric oxide may provide uterine relaxation to facilitate embryo implantation following fertilization. Cervical NOS III, localized in the mucus-secreting epithelium and smooth muscle cells, exhibited enzyme abundance during pro-oestrus and oestrus, probably indicating cervical preparation to facilitate sperm entry following mating. Vaginal NOS III, found in the stratified squamous epithelial lining and in smooth muscle cells, was maximal during oestrus and pro-oestrus, suggesting that nitric oxide may stimulate vaginal secretions. Differential expression of NOS III by different reproductive organs during the oestrus cycle suggests a role for nitric oxide in modulating reproduction.     

Effects of progesterone and oestradiol on RNA and protein metabolism in the genital tract and on survival of embryos in the ovariectomized ewe.

The hormonal regulation of metabolism in the genital tract and the development of embryos during early pregnancy in the ewe have been examined. Ovariectomized ewes received injections of maintenance progesterone, oestrous oestradiol and priming progesterone according to schedules designed to simulate endogenous ovarian secretion during early pregnancy, around the time of oestrus and during the luteal phase of the oestrous cycle immediately preceding oestrus. The survival and development of embryos was dependent upon the dose of maintenaince progesterone and the duration of treatment at the time of transfer, but changes in progesterone dose did not change endometrial protein or RNA metabolism on particular days. Both priming progesterone and oestrous oestradiol were required for normal embryo development. Priming progesterone and oestrous oestradiol each increased endometrial RNA/DNA ratios during early pregnancy. There were no interactions between priming progesterone and oestrous oestradiol, their effects being simply additive. Neither maintenance nor priming progesterone had any effect on protein and RNA metabolism in the oviduct. It is suggested that in the intact ewe oestrogen secreted at oestrus and progesterone secreted prior to oestrus play important roles in the establishment of a uterine environment suitable for the subsequent normal development of embryos.     

Follicular dynamics and temporal relationships among body temperature, oestrus, the surge of luteinizing hormone and ovulation in Holstein heifers treated with norgestomet

The effects of chronic treatment with norgestomet on follicular dynamics, corpus luteum growth and function as well as the temporal relationships among body temperature, oestrous behaviour, the luteinizing hormone (LH) surge and ovulation following implant removal were studied in 16 Holstein heifers. Oestrous cycles of the heifers were initially synchronized using 2 injections of prostaglandin F-2 alpha (PGF-2 alpha) 12 days apart. The heifers were then implanted with a norgestomet ear implant for 9 days, beginning either at the middle of the synchronized cycle (dioestrus) or at the end of the synchronized cycle (pro-oestrus). Follicular dynamics, corpus luteum growth and regression, and plasma progesterone were not affected by norgestomet treatment at dioestrus. The dominant follicle present at the time of norgestomet implantation in the pro-oestrus group was maintained during the 9-day implant period of 6 of 8 heifers and ovulated after implant removal. Time from implant removal to onset of standing oestrus and time to LH peak following implant removal were highly correlated with the time of ovulation (r = 0.92 and 0.96, respectively). Onset of standing oestrus and the LH peak and the onset of standing oestrus and peak vaginal and rectal temperatures were also highly correlated (r = 0.96, 0.82 and 0.81, respectively). It is concluded that any decrease in pregnancy rates following treatment with norgestomet is not due to asynchrony among oestrus, the LH surge and ovulation.     

The effect of intrauterine and cervical manipulation on the equine oestrous cycle and hormone profiles.

Endometrial biopsy or endometrial biopsy and uterine culture taken on Day 4 after oestrus induced lysis of the corpus luteum (CL), resulting in a sharp decline in serum progesterone concentration and shortened the interoestrous interval in 8/12 and 32/33 oestrous cycles, respectively, during 2 experiments. Cervical dilatation 4 days after oestrus shortened the interoestrus interval in 5/10 and 0/5 oestrous cycles. Endometrial biopsy and culture on Days 1 and 3 after oestrus also induced CL lysis during 4 of 7 cycles. Total oestrogen (oestrone plus oestradiol) concentrations increased at the onset of the subsequent oestrus in mares biopsied on Day 4 of dioestrus or in control cycle oestrous periods. Endometrial biopsy also induced lysis of the CL in mares with persistent luteal function. It is postulated that intracervical or intrauterine manipulations during the luteal phase of the oestrous cycle may directly, or indirectly, stimulate the release of an endogenous luteolysin (prostaglandin) resulting in CL regression, followed by oestrus and ovulation in the mare.     

An investigation of the uterine luminal environment of non-pregnant and pregnant pony mares.

Uterine flushings were collected from 30 non-pregnant Pony mares on Days 8, 12, 14, 16, 18 or 20 after oculation. Mares were allowed a recovery period of one oestrous cycle and were mated at the next oestrus. They were then ovario-hysterectomized on days which corresponded to the day of the oestrous cycle to which they were assigned. Uterine flushings were analysed for total recoverable protein and acid phosphatase activity. Least squares analysis indicated a status X day interaction for total protein (P less than 0.10) and acid phosphatase activity (P less than 0.005) in which the latter was higher in uterine flushings during pregnancy. Peripheral plasma oestrone and oestradiol concentrations were measured by radioimmunoassay and results indicated that plasma oestrone concentrations in pregnant and non-pregnant mares were not different, and oestradiol was lower (P less then 0.005) in the peripheral plasma during pregnancy. conceptus membranes were incubated in vitro for 120 min in a chemically defined medium. Incubation medium was then assayed to assess oestrone and oestradiol production capacilities at Days 8, 12, 14, 16, 18 and 20 of pregnancy. Conceptus membrane production of oestradios (pg/5 ml/h) increased (P less than 0.05) from Day 8 (243 pg/5 ml) to Day 20 (108 763 pg/5 ml). A similar trend, but of lower magnitude, existed for oestrone production.     

Oestradiol and progesterone: soluble receptor levels and metabolism in the uterus of the ovariectomized ewe

Ovariectomized ewes received injections designed to mimic to some extent oestradiol and progesterone secretion during early pregnancy (maintenance progesterone), during oestrus (oestrous oestradiol) and during the luteal phase of the previous cycle (priming progesterone). The animals were killed at times equivalent to 1, 4 or 7 days after oestrus in those animals which had received oestrous oestradiol. The level of soluble oestradiol and progesterone receptors in whole uterus, and [3H]oestradiol and [3H]progesterone metabolism by uterus minces were measured. Oestradiol receptor level was highest on day 1 in those animals receiving oestrous oestradiol with no significant effect at any stage of the inclusion or omission of priming or maintenance progesterone. Progesterone receptor level was also high on day 1 in those animals receiving oestrous oestradiol with high levels maintained to day 4. Again, inclusion of priming or maintenance progesterone was without effect. In animals not receiving oestrous oestradiol the level of both receptors was uniformly low. Metabolism of [3H]oestradiol was low and not affected by treatment. [3H]Progesterone metabolism, although more variable, was also low and not affected by treatment.     

Changes in neurohypophysial cholecystokinin content during oestrous cycle in the rat

Cholecystokinin content in the neurointermediate lobe of the rat pituitary was measured by radioimmunoassay during the different stages of the oestrous cycle. Higher levels were observed in pro-oestrus and oestrus than in metoestrus and dioestrus rats.

This difference is similar to the variation observed in the same circumstance concerning oxytocin in the neurohypophysis and neurosecretory activity in magnocellular neurons. These results are discussed in relation to the coexistence of oxytocin and cholecystokinin in neurons of the hypothalamoneurohypophysial system.     

Some aspects of immunology of the bovine uterus related to treatments for endometritis

Endometritis in breeding cattle occurs during the postpartum period, and is associated primarily with contamination of the reproductive tract involving Arcanobacter pyogenes (formerly Actinomyces pyogenes) together with Gram-negative anaerobes. Polymorphonuclear inflammatory cells (PMNs) contribute partly to the defense mechanisms against micro-organisms contaminating the vagina and uterine lumen, whose phagocytic activity depends on bacterial opsonisation by humoral antibodies; significant numbers of lymphocytes are also present. Whilst leukocyte numbers in the uterine lumen are relatively high during metoestrus and dioestrus compared to other phases of the oestrous cycle, their functional activity is unaffected. Humoral antibody concentrations in the reproductive tract are stimulated following exposure to local antigen, and the response is site dependent; of the several different classes of immunoglobulins, IgG predominates in the uterus and IgA the vagina. Only a portion of the total IgG1 found on the uterine lumen is synthesised locally in the endometrium, the remainder and all of the IgG2 is derived from the local uterine blood supply. Generally, concentrations of immunosuppressant proteins present in the uterine lumen increase under progesterone dominance, and these inhibit lymphocyte proliferation, making the uterus more susceptible to infection. The relationship between uterine susceptibility to micro-organism contamination and the luteal phase of the oestrous cycle is still unclear. Intrauterine infusion of immunomodulators such as E. coli lipopolysaccharides (LPS) or oyster glycogen, in healthy cows and those with endometritis, stimulates leukocytes to migrate into the uterine lumen. At a dosage rate of 100 microg, lipopolysaccharides are not absorbed by the healthy endometrium and do not alter the oestrous cycle length. It is unknown, whether a similar dose can be absorbed through an inflamed endometrium in naturally occurring cases of endometritis to cause systemic illness. Currently, prostaglandin F2alpha is recommended for treating endometritis in both cycling and non-cycling cows, but its mode of action in non-cycling cows is not fully understood. The efficacy of endometritis treatment using an intrauterine infusion of an immunomodulator in cases occurring naturally has not been determined on a large scale.     

Exogenous progesterone reduces follicular prostaglandin E and 6-keto prostaglandin F-1 alpha in the cyclic hamster

In cyclic hamsters, exogenous progesterone (100 micrograms) administered s.c. at 09:00 h on the day of dioestrus II reduced prostaglandin (PG) E and 6-keto PGF-1 alpha but not PGF concentrations in preovulatory follicles measured at 09:00 h of pro-oestrus. The injection of 10 micrograms ovine LH (NIADDK-oLH-25) concurrently with 100 micrograms progesterone on dioestrus II prevented the decline in follicular PGE and 6-keto PGF-1 alpha values. Administration of LH alone did not significantly alter follicular PG concentrations. Inhibition of follicular PGE accumulation by progesterone was due to a decline in granulosa PGE concentration and not thecal PGE. Progesterone administration also reduced follicular oestradiol concentrations. Administration of oestradiol-17-cyclopentanepropionate (ECP) (10 micrograms) with progesterone did not prevent the decline in follicular PGE and 6-keto PGF-1 alpha but did increase follicular PGF concentrations. However, ECP given alone on dioestrus II reduced follicular PGE and increased PGF concentrations in preovulatory follicles on pro-oestrus. It is concluded that exogenous progesterone administered on dioestrus II inhibits granulosa PGE and 6-keto PGF-1 alpha accumulation in preovulatory follicles, probably by reducing serum LH concentrations, and that the granulosa cells, which are LH-dependent, are a major source of follicular PGE.     

In-vivo myometrial electrical activity in the cyclic mare

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.     

Influence of progesterone administration in early stage of development on morphometric parameters of the rat uterus

The aim of this study was to evaluate the effects of single injection of 0.5 mg progesterone in neonatal stage of development on morphometric parameters of the adult rat uterus. The results showed that there were no changes ofendometrial morphometry in respect to oestrous cycle in neonatal treated rats. Neonatal administration of progesterone decreased the myometrium thickness in oestrus and dioestrums owing to circular muscle layer, and disturbed sex steroids secretion during the oestrous cycle. These data suggest that neonatal administration progesterone reduces the endometrium sensitivity to sex steroids and produces the myometrium atrophy.     

Plasma concentrations of 13, 14-dihydro-15-keto prostaglandin F(2alpha) and progesterone during oxytocin-induced oestrus in the goat

Fertile oestrus was induced in dairy goats by sub-cutaneous administration of 100 i.u. oxytocin per day between days 3-6 of the oestrous cycle. Peripheral plasma concentrations of 13, 14-dihydro-15-keto-prostaglandin F(2alpha) (PGFM), the major metabolite of prostaglandin (PG) F(2alpha), were elevated significantly (P<0.001), relative to controls, 30 minutes after oxytocin with peak values of between 300-800 pg ml(-1). Unlike control animals, plasma progesterone concentrations did not rise in the oxytocin-treated group after day 4. These results lend support to the hypothesis that the luteolytic effect of oxytocin in goats may be mediated via uterine PG production.     

Matrilysin activity in the rat uterus during the oestrous cycle and implantation

The objective of this study was to follow changes in the activity of the small matrix metalloproteinase matrilysin (MMP-7) in the rat uterus during the oestrous cycle and embryo implantation. Matrilysin was extracted from rat uteri, partially purified and separated into active and latent forms. The two forms of the enzyme were quantified at all stages of the oestrous cycle and after oestradiol and progesterone treatment. The activity was also measured during the first 7 days of pregnancy. Both latent and active forms of MMP-7 reached a peak during the pro-oestrous stage of the cycle; the concentrations were three times higher than at dioestrus and metoestrus. In rats treated with 0.1 mg oestradiol at metoestrus, both latent and active forms of the enzyme increased by more than two-fold after 24 h. In rats treated at pro-oestrus with 0.4 mg progesterone, there was a 70% increase in latent MMP-7, but no change in the active form. The highest concentrations of MMP-7 were observed on the first day of pregnancy. Between days 3 and 7 of pregnancy, the concentrations were relatively constant and comparable to the low concentrations at dioestrus. Enzyme activities were not different at implantation sites compared with remote sites.