Motility of the oviduct and uterus of the cow during the oestrous cycle.

Recordings of electrical activity of the oviduct and uterus were obtained during three oestrous cycles in cows fitted with an extra-cellular multi-electrode assembly. The stages of the cycle were identified by the appearance of the cervico-vaginal secretions and changes in the peripheral plasma level of progesterone were determined by radioimmunoassay. A gradual transition from local non-propagating electrical activity to propagating electrical activity with increase in the duration of contractions and then of their amplitude occurred 48 hr before the onset of oestrus. The transition coincided with a rapid decrease in progesterone level from 5 to 10 ng/ml to less than 0-1 to 0-4 ng/ml. This phenomenon was recorded from all uterine electrode sites, but was most marked at the uterotubal junction. Two days before oestrus, trains of potentials and bursts of activity became progressively grouped, apparently randomly, into prolonged phases in the distal portion of the oviduct and over the entire myometrium. During oestrus, the phases of activity became synchronized at these sites and both their amplitude and frequency reached a maximum. The strength but not the frequency of the phases diminished progressively 3 days after oestrus, followed by relative inactivity. The last remaining zone of activity was the uterotubal junction. During oestrus, the activities of the oviduct and the uterus were modified by oxytocin and adrenaline, the effect of the former being more marked on the uterus and that of the latter on the oviduct.     

Fertilization rate after synchronization of the oestrous cycle in heifers with prostaglandin F2alpha.

Oestrus was synchronized in 31 heifers by the intrauterine administration of PGF₂α than salt. Nineteen were given 2 doses of 0.5mg 24 hr apart, and 10 of these received 1500 I.U. of PMSG i.m. 24 hr before the treatment with PGF₂α. The remaining 12 heifers in the experiment were given a single dose of 2mg followed at the beginning of oestrus by 1500 I.U. of HCG i.m. Of 9 heifers which received only the two doses of 0.5mg (Group 1), 7 were observed to have corpora lutea when slaughtered 56–72 hr after the onset of oestrus, and four fertilized eggs were recovered. In those which received PMSG before the double injection of PGF₂α(Group 2), 118 corpora lutea were observed at slaughter and 34 fertilized eggs were recovered. Each heifer which received a single injection of PGF₂α and HCG had a corpus luteum, and 9 fertilized eggs were recovered. Unovulated follicles were most commonly observed in the PMSG-treated heifers but they were also observed in the heifers given the double injection treatment. It was observed that in the two-injection treatments, whether or not given PMSG, time of ovulation relative to the onset of oestrus was variable, and eggs were found in the uterus before the expected time.     

Patterns of oviducal motility in the cow during the oestrous cycle

Microtransducers sensitive to changes in internal diameter were chronically implanted in the oviducts of 5 dairy cows. Motility patterns were recorded throughout 9 oestrous cycles. Cyclic variations in patterns of motility were observed and compared with circulating concentrations of plasma progesterone. Luteal-phase motility patterns were of low amplitude and frequency. The frequency and amplitude of motility increased 3-5 days before behavioural oestrus. This activity consisted primarily of longitudinal muscle contractions, with an interspacing of circular muscle activity occurring during oestrus. Patterns of activity after oestrus were similar to those before oestrus, with activity decreasing 3-5 days after oestrus. Transducers implanted bilaterally in 2 animals permitted observation of asynchronous patterns between right and left oviducts. Preliminary data suggested a higher level of activity in the oviduct ipsilateral to the active ovary. These variations may be due to a local effect, possibly mediated by the functional ovary or the ovum.     

Differential transport of spermatozoa into the two sides of the genital tract of a monovular marsupial, the tammar wallaby (Macropus eugenii)

Ovulation in the tammar wallaby alternates between the ovaries. The genital duct of each side enters the median vaginal culs-de-sac separately. Post-partum oestrus occurred 0.4 days after birth and ovulation 1 day later. After a single copulation spermatozoa were found in both cervical canals at 0.5 h and extended to the oviduct on the non-parturient side only by 8 h. Very few spermatozoa were found in sections of the post-partum uterus or its associated oviduct at any time. Spermatozoa were recovered by flushing from both sides but the numbers were 2-20 times greater in the non-parturient than in the post-partum side: the greatest difference occurred in the cervical canals 2-5 h after copulation. In females which had undergone a previous infertile cycle, spermatozoa were abundant in both cervices and both uteri. It is concluded that the differential distribution of spermatozoa in post-partum animals was (1) due to failure of transport in the recently pregnant side of the tract, rather than attraction of spermatozoa to the ovulation side, and (2) established at the cervix which, on the ovulation side, provides a reservoir of spermatozoa for 24 h after copulation.     

The effect of oestrogen administered during the progestational phase of the cycle on transport of spermatozoa in ewes.

Two split-plot factorial experiments are described, the first with 72 entire cyclic ewes and the second with 80. The pattern of transport of spermatozoa through the reproductive tract was studied, following treatments with progestagen and oestrogen or with oestrogen alone during 2 weeks preceding insemination. A daily dose of 25 mug oestradiol-17 beta administered to ewes for 14 days preceeding oestrus had a deleterious effect on the passage of spermatozoa through the cervix into the uterus within the first 2 hr after insemination. The numbers of spermatozoa recoverable from the cranial region of the cervix 2 hr after insemination appeared to be related to the numbers in the oviducts at 24 hr. These numbers were related to fertility data from an earlier experiment using similar treatments. The data for log numbers of spermatozoa recoverable from the cervix formed a near-normal distribution and so were suitable for formal statistical analysis. There was an interaction between progestagen and oestrogen influence before mating on the pattern of sperm transport through the cervix.     

Influence of hCG injection and steroid treatment on prostaglandin metabolism by rabbit uterus and oviduct

Metabolism of PGE-2 and PGF-2 alpha by cytosolic fractions (100 000 g supernatant) of rabbit uterus, oviduct and lung was measured in vitro. Metabolism of PGE-2 was greater than that of PGF-2 alpha for oviduct and uterus. After an ovulating injection of hCG metabolism of both PGE-2 and PGF-2 alpha by lung and uterus declined linearly up to 72 h (during the time of ovum transport). The amount of PG metabolism by the oviduct did not change significantly during this period, but the percentage changes of PGE-2 and PGF-2 alpha metabolism from oestrous values did differ, and perhaps indicated a change in the ratio of intracellular PGs. No change of metabolism of either PG by lung, uterus or oviduct occurred at 24 or 72 h after an injection of 250 micrograms oestradiol cyclopentylpropionate given concomitantly with the hCG (a treatment regimen which causes 'tube-locking' of ova). However, progesterone treatment, in a regimen known to cause accelerated transport of ova through the oviduct, caused significantly enhanced metabolism of both PGE-2 and PGF-2 alpha by uterus and oviduct, but not lung, 30 and 48 h later except for PGE-2 by uterus at 30 h. These results suggest that changes in metabolism of PGE-2 and PGF-2 alpha by the oviduct may be involved in the mechanisms controlling ovum transport.     

Plasma concentrations of gonadotrophins, preovulatory follicular development and luteal function associated with bovine follicular fluid-induced delay of oestrus in heifers

In Exp. 1, injections of 10 ml bovine follicular fluid (bFF, i.v. or s.c.), given twice daily for 3 days after injection of a luteolytic dose of PGF-2 alpha, delayed the onset of oestrus in 3 of 6 heifers to 8 or 9 days after PGF-2 alpha, as compared with 2 or 3 days after PGF-2 alpha in control heifers. Mean plasma concentrations of FSH and LH during the injection period were not different from those in saline-injected heifers. In Exp. 2, i.v. injections of 20 ml bFF twice daily for 3 days uniformly delayed oestrus to 8 days after PGF-2 alpha (N = 4) and injections of 20 ml bFF i.v. every 6 h for 24h on the day of PGF-2 alpha injection delayed oestrus to 5.0 +/- 0.6 days after PGF-2 alpha as compared with 2.8 +/- 0.3 days for control heifers. In both treatment groups, plasma concentrations of FSH were suppressed during the injection period and increased transiently after treatment, but plasma concentrations of LH during the injection period were not different from those of control heifers. Plasma levels of oestradiol in heifers given bFF remained basal for 2 or 3 days after treatment, then increased several days before the delayed oestrus, in a manner similar to that in control heifers, and elicited normal preovulatory surges of LH and FSH. Plasma concentrations of progesterone and the length of the next oestrous cycle were normal, indicating formation of functional corpora lutea. Therefore, bFF treatments appear to delay oestrus by selectively suppressing plasma FSH, without affecting LH, and delaying the development of the preovulatory follicle. These results suggest that FSH may be critical to support the growth and development of the preovulatory follicle after luteolysis in cows.     

Fertility, ovulation and maturation of eggs in mares injected with HCG

Pony mares were observed from January to August for incidence of oestrus, duration of oestrus, length of the oestrous cycle and for ovulation and fertility after injection of HCG. From January to 15 May most mares showed oestrus but the duration of oestrus was quite variable and few mares ovulated in response to HCG. From 15 May to 17 August oestrous cycles were more regular and ovulation was induced within 40-50 h by an intramuscular injection of 1500-5000 i.u. HCG. Pregnancy was established by one mating at a fixed time after HCG in 20 of 69 mares. Degenerate eggs were recovered from the oviducts of anoestrous recently ovulated, mated, unmated and pregnant mares. The first polar body was formed before ovulation in 2 eggs and had not formed in 2 recently ovulated eggs flushed from the oviduct. The second polar body formed after sperm penetration 10-12 h after ovulation. After formation of pronuclei, the first cleavage division occurred at 20 h and the second at 32 h after ovulation. Oestrus was inhibited by progesterone administered by vaginal devices but occurred within 1-3 days in 12 of the 20 mares after withdrawal of the devices.     

Progesterone binding in rabbit oviduct and uterus.

Progesterone binding of high affinity with a dissociation constant of 10(-9) M was identified in cytosol of rabbit oviduct and uterus. Macromolecules with sedimentation coefficients of 7-8 S and 4-5 S were present. Progesterone receptor concentration was two to fivefold lower in the oviduct when compared with the uterus. The receptor concentration declined steadily from 3 hr until 144 hr after mating in the uterus; however, the decline in oviductal receptor was not significant until the sixth day of pregnancy. Progesterone receptor concentration in rabbit oviduct and uterus in estrus and early pregnancy was greater than estradiol receptor levels.     

Protein synthesis in the early stages of cardiac hypertrophy

Cardiac hypertrophy, induced in rats by either tri-iodothyronine or isoproterenol, administered daily for 7 days, was monitored using several parameters. Both treatments increased RNA concentrations 24 hr after the first injection, while heart weight increased following 2 injections to 46% above control after 7 days. Cardiac protein synthetic activity, as determined by the rate of peptidyl-puromycin formation, was increased by both tri-iodothyronine and isoproterenol 24 hr after a single injection, implying an increase in the number of functional ribosomes. RNA activity (the rate of peptidyl-puromycin formation per unit RNA) remained constant, suggesting that neither accelerated rates of initiation or translation nor increased activation of pre-existing, non-translating ribosomes was involved in the observed increase in protein synthetic activity. In contrast, constant infusion of [14C] tyrosine indicated no change in protein synthetic rate 24 hr after a single tri-iodothyronine injection and decreased protein synthetic rate after isoproterenol injection. It is concluded that the use of [3H]puromycin to estimate protein synthetic activity may be a more sensitive procedure for detecting early changes in protein synthesis in cardiac hypertrophy than constant isotope infusion, owing to the problems associated with determining the precise precursor pool for protein synthesis in this latter method.     

Proceedings: Action of chronically administered oestradiol on the uterus of the rat

Oestradiol induces increased synthesis of RNA and DNA in the uterus of ovariectomized rats. The effects of continuously administered oestradiol on nucleotide synthesis in the uterus of the rats are reported. Ovariectomized rats were given 2 Mug oestradiol-17 beta, subcutaneously, every 8 hr until autopsy at various times 1 to 7 days after the first injection of oestradiol. (3H) uridine or (3H) thymidine was administered intraluminally 15 min before beath. Uteri were processed for autoradiography. The number of labelled cells and the average number of grains/cell were counted. (3H) uridine labelling reached a peak at 6 to 54 hr and then decreased steadily thereafter. DNA synthesis was maximal at 48 hr in all regions and minimal at 144 hr. These results indicate that oestrogen caused maximum stimulation of RNA synthesis in the rat uterus at 30 and 48 hr respectively but activity was reduced thereafter. The uterine epithelium and stroma were hypertrophied and hyperplastic when RNA and DNA synthesis were minimal. This could be due to refractoriness of the specific target tissues to continued hormonal stimulation.     

Regulation by Gonadal Steroids of Estrogen and Progesterone Receptors Along the Reproductive Tract in Female Lambs

The regulation of estrogen and progesterone receptor (ER, PR) expression by estradiol (E2) and progesterone (P4) in the oviduct, uterus and cervix of female lambs was studied. The animals received three intramuscular injections of E2, P4 or vehicle with an interval of 24 h and they were slaugthered 24 h after the third injection. Determinations of ER and PR were performed by binding assays and mRNAs of ERα and PR by solution hybridization. High levels of ER and PR in both cervix and oviduct were found in the female lamb, differing from other mammalian species. No significant effects by either E2 or P4 treatment on ER and PR levels in the cervix and oviduct could be observed. E2 treatment increased the mRNA levels of ERa and PR more than 3-fold in the cervix, while P4 treatment increased the mRNA levels of ERa and PR in the uterus. The results show differential effects of gonadal steroids on sex steroid receptor expression along the reproductive tract in female lambs, suggesting that steroid target tissues can modulate responses to the same circulating levels of steroid hormones.     

Sequential Hormonal Changes in the Postpartum Dairy Cow

Peripheral plasma levels of 15-keto-13,14-dihydro-PGF2α, progesterone, Cortisol, LH and prolactin were studied in 6 primiparous postpartum dairy cows. The cows were followed by hormone measurements and clinical examinations from parturition until pregnancy was established. Blood was collected 3 times per day. The cervix, uterus and ovaries were examined by rectal palpation at 6–10 days intervals. The cows were observed for signs of oestrus twice daily and were additionally teased with a bull to provoke standing heat. Four cows had a normal parturition and dropped their fetal membranes shortly afterwards. (NR group). The remaining 2 retained their fetal membranes for more than 24 h following parturition (RFM group). One out of 6 cows showed standing oestrus at the first ovulation, 4 animals were in oestrus at the second ovulation and all cows showed signs of oestrus at the third ovulation. Although the length of the first luteal phase varied from 9 to 22 days a corpus luteum was in all cases palpated. The secretion of progesterone during the first luteal phase was terminated by a PGF2α release. A significant difference in 15-keto-13,14-dihydro-PGF2α levels between the 2 groups was found on days 0–4 (2.39 vs 6.87 nmol/1 at Ρ < 0.06). Postpartum prostaglandin F2α release as reflected by the level of 15-keto-13,14-dihydro-PGF2α lasted shorter in the NR group than in the RFM group (15–17 vs 21 days). Significant positive correlations between 15-keto-13,14-dihydro-PGF2α and Cortisol as well as between prolactin and Cortisol during the first 24 days postpartum were noted only in cows having normal parturition. The most pronounced daily prolactin variations occurred during the second luteal phase (NR group), when a significant difference between the times 8.00, 12.00 and 15.00 was recorded (14.7, 31.5 and 19.7 μg/l, respectively). Moreover, a partial negative correlation between log value of prolactin and arithmetical value of LH was found in these cows only during the first luteal phase after parturition.     

Electromyographic activity of the uterus in the ewe during the sexual season: comparison of natural estrus and estrus induced by progestagens alone or with PMSG supplementation

Electromyographic (EMG) activity of the uterus was recorded in vivo in 3 groups of ewes in oestrus during the breeding season. The ewes were either in natural oestrus or in oestrus induced by progestagens with or without PMSG supplementation. Plasma concentrations of LH were measured at regular intervals in order to determine the onset of the preovulatory surge of LH (To). During natural oestrus, the uterus exhibited a spontaneous rhythmic activity composed of bursts of potentials. Burst frequency was maximal at the moment of preovulatory LH release, then decreased. Percentages of downward, upward and non-definite propagations were calculated during 1-h periods at the beginning of the LH peak and 6, 12, 24 and 36 h afterwards. There was no difference between the relative proportions of these types of propagation at any of the times analysed. Mean burst frequency was not modified by progestagen pre-treatment but showed more interanimal variability at the moment of maximal frequency in ewes treated with progestagen. For the first 12 h after To, the organization of uterine activity was not modified by progestagens but non-definite propagation became predominant 24 and 36 h after To, namely from the time of ovulation onward. PMSG increased burst frequency. The increase of frequency variability between animals, normally observed at the beginning of oestrus, and the predominance of non-definite propagation, normally observed at the end of progestagen-induced oestrus, did not appear when the ewes had been treated with PMSG.     

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.     

Osteopontin improves in vitro development of porcine embryos and decreases apoptosis

An optimal environment for fertilization and early embryonic development is provided by the mammalian oviduct and uterus. The secretory cells lining the lumen of the oviduct and uterus synthesize and secrete proteins that have been shown to interact with and influence the activities of gametes and embryos. Western blotting in this study demonstrated that a 50-kDa secreted phosphoprotein 1 (SPP1) form was present in the uterus on Days 0, 3, and 5 in pregnant and nonbred gilts, and the concentration of SPP1 on Day 0 was higher than on Days 3 and 5 in pregnant gilts, but in nonbred gilts the concentration of SPP1 on Day 0 was higher than Day 3, but not Day 5. In addition, we show that addition of 0.1 microg/ml SPP1 to the culture medium after fertilization increased the percent cleaved (24 hr: 23.6 +/- 1.29(a) vs. 18.7 +/- 0.65(b) (2-cell %)), and the percent blastocyst (37.2 +/- 1.12(a) vs. 30.9 +/- 0.56(b)) derived from IVF (P < 0.05). In parthenogenetic-derived embryos the percent cleaved was increased due to SPP1 at 24 hr (24.0 +/- 1.59(a) vs. 19.7 +/- 1.59(b) (>2-cell %)), and at 48 hr (72.9+/- 2.99(a) vs. 63.3 +/- 2.99(b)), but not the percent blastocyst. By TUNEL assay, SPP1 decreased both apoptosis (7.9 +/- 0.04(a) vs. 13.1 +/- 0.02(b)) and the percent fragmentation (45.2 +/- 0.07(a) vs. 58.8 +/- 0.03(b)). We conclude that SPP1 can improve development in vitro possibly by reducing the rate of apoptosis.     

重组禽腺联病毒介导的输卵管暂态生物反应器的初步研究

将输卵管特异表达启动子调控的人组织激肽释放酶（human tissue kallikrein,hKLK1）表达盒插入至AAAV转移载体pAITR中,与AAAV包装载体pcDNA-ARC及腺病毒辅助质粒pHelper三质粒利用磷酸钙沉淀法共转染AAV-293细胞,制备输卵管特异表达hKLK1的rAAAV。将获得的重组病毒以每只鸡2×1010病毒颗粒数翅静脉注射正常产蛋母鸡,RT-PCR结果显示hKLK1只在输卵管部位表达;酶活性检测结果表明：注射后第2天就可以检测到rhKLK1的活性,第3周表达量最高,达107.3U/ml,表达时间持续6周之久;用含rhKLK1的蛋清灌喂自发性高血压大鼠（SHR）,可使其血压下降70mmHg,5天后回升到饲喂前水平。以上结果表明重组禽腺联病毒介导的鸡输卵管暂态生物反应器不仅具有很好的组织特异性,而且可指导外源基因长期稳定的表达。     

Requirement for progesterone priming and its long-term effects on implantation in the mouse

At least 48 hr of progesterone (P4) priming has been documented to be essential for P4 and estrogen to initiate implantation in the rat. However, the length of this P4 priming requirement for implantation in the mouse has not been experimentally defined. Therefore, our first objective was to determine the length of P4-priming requirement for implantation in the mouse. Day 4 blastocysts were transferred into the uteri of Day 5 or Day 6 pseudopregnant mice that were ovariectomized on Day 1 (= vaginal plug) and treated with a single injection of P4 and 17 beta-estradiol (E2) only on Day 5, or a single injection of P4 on Day 5 followed by a second injection of P4 plus E2 on Day 6, respectively. Although none of the transferred blastocysts implanted in the uteri of P4-unprimed recipients, 46% of the transferred blastocysts implanted into the uteri of all recipients that were first primed with P4 24 hour prior to a second injection of P4 and E2. These results suggest that in contrast to the rat, the mouse uterus requires at most 24 hr of P4 priming before P4 and estrogen can initiate implantation. Our second objective was to determine whether P4 priming has a long-term effect on implantation in the mouse. Our present results and those of others suggest that the mouse uterus is exposed to rising P4 levels for 24 hr prior to implantation on Day 4 of pregnancy. Therefore, in the present investigation, induction of implantation by an injection of P4 and E2 following 5 days of ovariectomy performed on Day 4 of pregnancy clearly suggests that once exposed to P4 for 24 hr, the mouse uterus retains a long-term effect, i.e., following P4 withdrawal for several days, 24 hr of initial P4 priming is no longer required for P4 and estrogen to initiate implantation. Our next objective was to explore whether this long-term effect of P4 priming on implantation can be prolonged and potentiated by increasing the length of initial P4 priming. Thus, when the mice were ovariectomized on Day 4 of pregnancy and treated with P4 beginning on Day 5 for 4 days, the long-term effect on implantation was prolonged (8 days vs 5 days following P4 withdrawal) and potentiated (94% vs 0% mice with implantation following 8 days of P4 withdrawal) as compared with those with no P4 priming after ovariectomy.(ABSTRACT TRUNCATED AT 400 WORDS)     

Oestrous synchronization in cattle using progesterone and a PGF analogue

A regime consisting of in jections of 150mg and 100mg of progesterone subcutaneously (s.c.) 3 days apart, followed 2 days later by a single injection intramuscularly (i.m.) of a PGF analogue (ICI 80996), and after further 12 hr interval, 1000 i.u. PMSG s.c. was used to synchronize oestrous periods in 16 cows. Eight cows (50%) conceived after insemination with frozen semen after synchronized oestrus; 14 cows came into oestrus over a 36-hr period.     

Cycles of mating behaviour, oestrogen and progesterone in the thick-tailed bushbaby (Galago crassicaudatus crassicaudatus) under laboratory conditions

Vaginal smears and blood samples were taken throughout the reproductive cycle of female Galago c. crassicaudatus. Blood plasma was assayed for oestradiol and progesterone, and vaginal smears were initially classified dioestrus or vaginal oestrus. During vaginal oestrus the females were tested daily for sexual receptivity by being placed with a male. Those days on which the male achieved intromission were reclassified as behavioural oestrus. During dioestrus the females were tested weekly with males. Female receptivity increased and then declined across a 6-day period of behavioural oestrus during the 44-day cycle. Fully cornified smears were characteristic of the period of maximal receptivity and oestradiol secretion. The luteal phase lasted 24 days with a plasma progesterone peak midway through dioestrus.