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Effect of boar exposure at time of insemination on factors influencing fertility in gilts.

The effect of boar exposure during artificial insemination (AI) on semen backflow, fertilization, and embryo quality was evaluated. Gilts (approximately 170 d) were induced into estrus with PG600, and ovulation was synchronized using hCG 72 h later. Estrus detection was initiated after PG600 and continued at 12-h intervals. At estrus, gilts were allotted to receive boar exposure (BE, n = 20) or no boar exposure (NBE, n = 20) during AI. Gilts receiving NBE were identified to be in estrus prior to AI and the boar was then removed for 1 h, whereas gilts in the BE group received 15 min of exposure during AI. Insemination occurred in crates at 12 and 24 h after onset of estrus with 3 x 10(9) sperm/80 mL. Backflow was collected continuously with samples taken at time 0, (during AI), and at 0.25, 0.5, 0.75, 1, 2, 4, and 8 h after first and second AI. The effect of treatment was evaluated for time of insemination (min), backflow (mL), and sperm in backflow samples. Oviducts were flushed 2 d after first AI to evaluate the effect oftreatment on fertilization rate, accessory sperm numbers on embryos (scored 1 to 5), and embryo quality. There was no effect of first or second AI; therefore, data were pooled. Average duration of AI was 3.7 +/- 0.2 min and was not influenced by BE (P < 0.10). However, during the initial stage of AI, BE reduced the volume of semen (18.6 vs 32.4 +/- 3 mL) and the number of sperm lost (0.8 vs 1.3 +/- 0.15 x 10(9) sperm) compared to NBE (P < 0.05). There was a treatment x time effect (P < 0.05) for volume of backflow. By 45 min, the BE gilts lost more volume (9.0 vs 3.6 mL) compared to the NBE group, but sperm loss did not differ. Between 1 and 8 h after AI, neither volume nor sperm loss was influenced by treatment. By 8 h, total leakage (65 vs 63 mL) and total sperm loss (1.6 x 10(9) vs 1.8 x 10(9) sperm) were not influenced by BE (P > 0.10). However, more accessory sperm (P < 0.01) were found on embryos for the NBE (> or = 11 sperm/embryo) compared to BE embryos (< or = 10 sperm/embryo). Despite this observation, percentages of fertilized embryos (99.5 +/- 0.5 %) and number of embryos (11.5 +/- 0.1) were not different (P > 0.10). In conclusion, AI in the presence of a mature boar did not affect total semen leakage, sperm loss, fertilized embryos, or embryo quality. The importance of boar exposure during insemination was evident from less leakage during insemination, but had no effect on fertility; this suggests that the elimination of boar exposure during AI may not be deleterious to reproductive performance.

Animals↗

[Analysis of platelet-activating factor and its effect on the lung].

One of the lipid mediators, platelet-activating factor (PAF), is reported to be involved in a variety of biological phenomena including not only harmful reactions such as allergy or inflammation, but also physiological phenomena like nerve cell differentiation, sperm mobilization, ovulation, implantation, parturition etc. We reviewed the methods of detecting PAF in biological fluids and the effects of PAF on fetal lung maturation and pulmonary diseases. Fetal lung has a capacity to produce PAF and this autacoid is involved in glycogen breakdown to furnish both glycerol backbone, such as dihydroxyacetone phosphate and glycerol-3-phosphate, and the fatty acids utilized to synthesize pulmonary surfactant. PAF also enhances secretion of pulmonary surfactant. However, PAF causes eosinophil recruitment to the lung, activation of eosinophil and neutrophil, bronchoconstriction, and lung edema, and increases bronchial hyperreactivity. All are characteristics of the pathophysiology of bronchial asthma. We examined PAF acetylhydrolase activity in plasma from patients with bronchial asthma, and found the activity was significantly lower in severe cases than that in mild or moderate cases. Ketotifen had no effect on this enzymatic activity. Evaluating PAF acetylhydrolase activity may help determine the severity of bronchial asthma. PAF is also involved in chronic lung disorders of newborns, such as bronchopulmonary dysplasia. We treated a 3-month-old child with frequent pulmonary problems who was made a diagnosis of bronchopulmonary dysplasia during neonatal period. After administration of Ketotifen for one and a half months, his clinical symptoms improved dramatically.

Animals↗

The effect of repeated boar exposure on cortisol secretion and reproduction in gilts.

It has been proposed that short-term activation of the hypothalamo-pituitary adrenal axis, with a consequent increase in the secretion of cortisol, amy disrupt the endocrine events prior to ovulation and thereby impair reproduction in females. We investigated this concept in gilts in which oestrus was detected by introduction to boars, where intense physical contact is possible, or by applying pressure to the back of gilts (back-pressure test) during fence-line exposure to boars, where intense physical contact is prohibited. We expected that there would be a greater release of cortisol and that reproduction would be inhibited in gilts introduced to boars compared to gilts in which the back-pressure test was used. As expected, introduction of gilts to boars resulted in a significant transient increase in plasma concentrations of cortisol while there was no significant effect of using the back-pressure test on plasma cortisol. Nevertheless, introduction of gilts to boars did not impair reproduction and there was no effect of method of detecting oestrus on duration of oestrus, sexual receptivity, fertility or fecundity. The length of the oestrous cycle was decreased and ovulation rate increased in gilts that were introduced to boars compared to gilts that underwent the back-pressure test, indicating that introduction of gilts to boars may have stimulated these aspects of reproduction. These stimulatory effects may have been due to an increased exposure of gilts to sexual behaviour and stimuli from boars when introduced to boars and/or to stimulatory effects of the hypothalamo-pituitary adrenal axis on some aspects of reproduction.

Adrenal Glands↗

Effect of cohabitation with white-faced ewes on estrous activity of Hampshire and Suffolk ewes exposed to rams in June.

Two groups of 24 Hampshire and 26 Suffolk purebred ewes each were used to study effects of cohabitation with cyclic white-faced (WF) ewes on estrous activity in June. Ewes lambed in January, February and March and had been isolated from mature rams since the previous fall breeding. From June 1 to July 2, treated (T) ewes were exposed to vasectomized rams and to 65 WF ewes; control (C) ewes were exposed only to vasectomized rams. Ovulation was assessed with biweekly serum progesterone assays; crayon marks were used to detect estrus. Daily observations of ram behavior were conducted to assess sexual activity of rams joined with T and C ewes. Cohabitation with WF ewes increased (P less than .01) ovulation percentages from 46% in C (42% for Hampshires and 50% for Suffolks) to 76% in T ewes (79% for Hampshires and 73% for Suffolks). Mating percentage also was increased (P less than .05) by cohabitation with WF ewes from 14% for C ewes to 30% for T ewes. Rams with T + WF ewes spent more (P less than .05) time checking ewes for estrus than did rams with C ewes. Hence, cohabitation with cycling WF ewes increased ovulation and mating percentages. Many acyclic T ewes first ovulated after 10 or more days of teasing, possibly due to increased ram contact in the presence of WF ewes.

Animals↗

Oocyte-specific expression of mouse Zp-2: developmental regulation of the zona pellucida genes.

The zona pellucida surrounds all mammalian oocytes and plays a vital role at fertilization and in early development. The genes that code for two of the mouse zona proteins (ZP2 and ZP3) represent a developmentally regulated set of genes whose expression serves as markers of mouse oocyte growth and differentiation. We previously characterized the single-copy Zp-3 gene and showed that its expression is oocyte specific and restricted to a narrow window of oocyte development. We now define the Zp-2 gene transcript and show that it is coordinately expressed with Zp-3 only during the 2-week growth phase of oogenesis that occurs prior to ovulation. Like Zp-3, the expression of Zp-2 is restricted to oocytes, and, although not detectable in resting oocytes, both ZP2 and ZP3 transcripts accumulate to become very abundant messengers in 50-microns-diameter oocytes. Ovulated eggs contain ZP2 and ZP3 transcripts which are 200 nucleotides shorter than those found in growing oocytes and have an abundance of less than 5% of the peak levels. In an attempt to understand the molecular details associated with the developmentally regulated, tissue-specific gene expression of the zona genes, the Zp-2 genetic locus has been characterized and its 5' flanking sequences have been compared with those of Zp-3. Both genes contain three short (8- to 12-base-pair) DNA sequences of 80 to 88% identity located within 250 base pairs of their transcription start sites.

Amino Acid Sequence↗

Characterization of HoMADS 1 and its induction by plant hormones during in vitro ovule development in Hyacinthus orientalis L.

To understand the molecular mechanism of ovule development, a MADS box gene, HoMADS 1 , has been isolated from the ovule tissues of Hyacinthus . Sequence comparison showed that HoMADS 1 is highly homologous to both class C and D genes. Furthermore, phylogenetic analysis suggests that HoMADS 1 is most likely a class D MADS box gene. RNA hybridization revealed that HoMADS 1 was exclusively expressed in the ovules. Over-expressing HoMADS 1 in transgenic Arabidopsis plants produced ectopic carpelloid structures, including ovules, indicating that HoMADS 1 is involved in the determination of carpel and ovule identities. Interestingly, during in vitro flowering, no HoMADS 1 mRNA was detected in the floral tissues at high level hormones in the media. However, HoMADS 1 mRNA accumulated in the floral tissues when the regenerated flowers were transferred to the media containing low level hormones which could induce in vitro ovule formation. Our data suggest that the induction of HoMADS 1 by plant hormones may play important roles during ovule initiation and development in the regenerated flower. Whether HoMADS 1 expression is also regulated by cytokinin and auxin during ovule development in planta remains to be investigated.

Amino Acid Sequence↗

Knee joint kinaesthesia and neuromuscular coordination during three phases of the menstrual cycle in moderately active women.

An increased incidence of sports related injuries in the premenstrual phase as well as in the menstrual phase of the menstrual cycle has been described. This may be explained by alterations in proprioception and neuromuscular coordination due to hormonal variations. Prospective, within women analysis of knee joint kinesthesia and neuromuscular coordination were performed by repeated measures analysis of variance in three hormonally verified phases of three consecutive menstrual cycles. Thirty-two healthy, moderately active female subjects volunteered to participate in the study. Twenty-five of the subjects performed at least one hormonally verified menstrual cycle. A specially designed device was used to investigate knee joint kinaesthesia and neuromuscular coordination was measured with the square hop test. These tests were carried out in the menstrual phase, ovulation phase and premenstrual phase determined by hormone analyses in three consecutive menstrual cycles. An impaired knee joint kinaesthesia was detected in the premenstrual phase and the performance of square hop test was significantly improved in the ovulation phase compared to the other two phases. The results of this study indicate that the variation of sex hormones in the menstrual cycle has an effect on performance of knee joint kinaesthesia and neuromuscular coordination.

Adult↗

Expression of oestrus and timing of ovulation in pigs.

Oestrus is the period around ovulation in which sows show a standing response for boars, thus allowing the boars to mate with the sows. The duration of oestrus may vary between sows from 24 h up to 96 h, which is not related to systemic oestrogen concentrations. The moment of ovulation after onset of oestrus may vary from 10 h to 85 h. Consequently, onset of oestrus is not a good predictor for the time of ovulation. A reliable prediction of ovulation time would be worthwhile, since fertilization results are highly dependent on the moment of insemination relative to the moment of ovulation. However, at present, the duration of oestrus is the best 'predictor' of ovulation time and is independent of the duration of oestrus; ovulation takes place on average 70% of the way through oestrus. The following factors have been found to influence the duration of oestrus (and consequently the optimal time for insemination): boar-stimuli, stress, interval from weaning to oestrus, season and parity. Further study is needed to determine the causes of the variability in oestrus expression between sows, including the regulation of oestrus behaviour at the level of the brain.

Animals↗

Studies on the pattern of circulating steroids in the normal menstrual cycle. I. Simultaneous assays of progesterone, pregnenolone, dehydroepiandrosterone, testosterone, dihydrotestosterone, androstenedione, oestradiol and oestrone.

In an attempt to analyze the multiple changes and interactions in circulating steroid levels in the peri-ovulatory and peri-menstrual periods, the plasma levels of immunoreactive luteinizing hormone (LH), progesterone and unconjugated pregnenolone, dehydroepiandrosterone, testosterone, oestradiol and oestrone were assayed daily during a complete cycle in 17 normally menstruating women. In 14 of the 17 subjects studied androstenedione and unconjugated dihydrotestosterone were also estimated. The day of the LH-peak and the first day of menstruation, respectively, were used to synchronize the peri-ovulatory and peri-menstrual plasma levels of the various steroids. With the exception of dehydroepiandrosterone and dihydrotestosterone, the plasma levels of all steroids exhibited significant, but different changes during the cycle. Testosterone levels showed a slight but significant increase around the LH-peak, whereas the levels of pregnenolone and androstenedione were higher in the post-ovulatory than in the pre-ovulation periods. The levels of oestradiol and oestrone, as well as the ratios of oestradiol to oestrone gradually increased from the low values observed in the early proliferative phase to pre-ovulatory peak values. The relationship between peaks of oestradiol and oestrone and that of LH exhibited great individual variation. The same was true for the individual oestradiol to oestrone ratios. The combination of several steroidal signals did not improve the predictive value of the analyses. However, an increase of individual progesterone values by at least 0.35 ng/ml from the day preceding the LH-peak to the day of the LH-peak was observed in 13 of the 17 subjects. It is suggested that for the early detection of the LH surge and prediction of the subsequent ovulation daily assays of plasma progesterone are of more value than the assay of the other steroids investigated.

Adult↗

Ovulation rate and the concentrations of gonadotrophins and metabolic hormones in ewes infused with glucose during the late luteal phase of the oestrous cycle.

The positive relationship between nutrition and ovulation rate was investigated in sheep infused intravenously with glucose. Ovulation rate increased (2.0 +/- 0.0 vs 2.4 +/- 0.3) when ewes were given an infusion of glucose (60-65 mM/h) for five days in the late luteal phase of the oestrous cycle. The effect of glucose was obtained without any significant change in LH secretion. The concentration of FSH in glucose-infused ewes was lower during the infusion (luteal phase) but higher during the early follicular phase. These data suggest that the change in ovulation rate occurred without increased gonadotrophin support to the follicle during the late luteal phase, which is the period of the sheep oestrous cycle during which improved nutrition increases ovulation rate. There were no changes in GH or prolactin, but changes in circulating glucose and insulin levels were detected. We conclude that insulin, because of its role in cell growth and metabolism, is involved in mediating ovulation responses to nutritional stimuli, either directly or more likely by the stimulation of insulin-mediated glucose uptake.

Animals↗

Changes in serum and ovarian steroids during reproductive development in the female guinea pig.

This study was designed to measure ovarian hormones prior to and during the first estrous cycle in guinea pigs. Blood was obtained from 12 animals throughout the first estrous cycle. Ovaries and peripheral serum were obtained from 25 additional animals at various stages of development prior to and after first ovulation. Estradiol, progesterone, androstenedione, and testosterone were measured in all sera and half of the ovaries. The remaining ovaries were fixed for histology. Serum estradiol was nondetectable until a few days before first ovulation, but was present in the ovary throughout development. Serum progesterone was nondetectable until the day of ovulation, but the luteal phase pattern was similar to that observed in adults. Serum androgens were detectable throughout development, with androstenedione higher than testosterone. The immature ovary contained more testosterone than androstenedione, but this pattern was reversed after ovulation. These results indicate that the immature ovary in the guinea pig contains minimal amounts of estradiol and progesterone, the first estrous cycle is similar to that in adults, and that the pattern of ovarian androgen content changes during the peripubertal period.

Androstenedione↗

Effect on oestrus and ovulation of exposing creole goats to the male at three times of the year.

The characteristics of the oestrous and ovarian cycles of local goats were studied over 2 years, on 96 females in March, 60 in July and 100 in November. After 3 weeks of separation between sexes, the females with inactive ovaries were identified and harnessed males were introduced. Detection of oestrus was undertaken daily and the date and rate of ovulation were checked at laparoscopies. The proportion of females with non-cyclic ovaries before the mating period varied according to the season and whether the females were nulliparous or suckling. Among the females with inactive ovaries before the mating period, 97% ovulated 2 . 8 days after the introduction of males; 68% of these initial ovulations were accompanied by oestrus and 76% resulted in short luteal phases (5 . 3 days) followed by a second ovulation accompanied by oestrus 89% of the time. The proportions of initial ovulations with oestrus and of initial ovulations followed by a short luteal phase were in linear relationship with the proportion of females with non-cyclic ovaries before the mating period. For the females with cyclic ovaries before the mating period, the distribution of first oestrus during the 8 days after the introduction of males was significantly different from the expected uniform distribution. A possible luteolytic action of teasing was suspected. The fertility of females in all groups was high (greater than 79%). Litter size was not different for non-cyclic and for cyclic females.

Animals↗

Expression of the TAL1/SCL transcription factor in physiological and pathological vascular processes.

The TAL1/SCL transcription factor is essential for haematopoietic commitment and vascular remodelling during embryonic development. To help clarify its role in postnatal vascular processes, we characterized the expression of mouse Tal1 protein by immunocytochemistry in several experimental models of blood vessel formation. In adult mice, Tal1 protein was expressed in rare microvascular endothelial cells and in extravascular cells provisionally identified as endothelial progenitors from their morphology, proximity to vessels and expression of vascular endothelial growth factor receptor-2. The number of Tal1-expressing endothelial cells increased significantly but transiently in all the models-hormone-induced ovulation, wound healing and tumour development. Finally, Tal1 protein was detected in the nuclei of newly formed lymphatic endothelial cells in tumour-bearing animals. These results show that TAL1 is expressed by vascular endothelial cells and endothelial progenitors at sites of physiological and pathological neovascularization and suggest a role for this transcription factor in adult vasculogenesis. This work also provides the first evidence for TAL1 expression in lymphangiogenesis.

Animals↗

Beta-carotene uptake and changes in ovarian steroids and uterine proteins during the estrous cycle in the canine.

The uptake of beta-carotene by reproductive tissues and the effects of beta-carotene on reproductive function in the dog are unknown. We studied the uptake of beta-carotene by blood, corpus luteum, and uterine endometrium and the role of dietary beta-carotene in influencing ovarian steroid and uterine protein production during the estrous cycle in the dog. Mature female Beagle dogs (n = 56) were fed diets containing 0, 2, 20, or 50 mg of beta-carotene daily for approximately 6 wk before estrus detection. Blood was sampled at regular intervals from estrus through d 45 after ovulation (d 0 = ovulation), when laparotomy was performed. The ovaries were obtained for the isolation of corpus luteum. The uterus was flushed with phosphate-buffered saline and the endometrium obtained by scraping. Beta-carotene was not detectable in plasma, corpus luteum, or endometrium of unsupplemented dogs. However, beta-carotene and alpha-carotene in plasma, corpus luteum, and uterine endometrium increased in a dose-dependent manner. Alpha-carotene made up a high percentage of total carotenoids even though the alpha-carotene content in the dietary source was very low. Dogs fed 50 mg of beta-carotene had significantly higher concentrations of plasma progesterone between d 12 and 26 compared with unsupplemented dogs. Dietary beta-carotene did not influence plasma estradiol-17beta and total uterine proteins. Therefore, beta-carotene is absorbed into plasma, corpus luteum, and uterine endometrium of dogs. Furthermore, dietary beta-carotene increased plasma progesterone concentrations during the estrous cycle. It is possible that dietary beta-carotene may improve reproductive function in the canine.

Animals↗

Evaluation of response to hormonal therapy in prepubertal gilts of different genetic lines.

Research was conducted to determine the effect of genetic line on hormonally induced puberty. Two studies were conducted, the first to evaluate estrus response (n = 120 gilts) and the second to evaluate follicular development (n = 24 gilts). Gilts were allotted to treatments in a 2 x 2 factorial arrangement. Gilts from two genetic lines (Age at Puberty x Yorkshire [APY] and Relaxed Selection x Yorkshire [RSY] received either P.G. 600 [symbol: see text] (P) or no injection (C). The percentage in estrus within 5 d after treatment was greater (P < .05) for P than for C gilts. However, no difference was detected for the percentage of P or C gilts that had ovulated within 14 d. For the APY line, P gilts had greater (P < .05) ovulation rates than C gilts. However, for the RSY line, P and C gilts did not differ in ovulation rate. More P gilts than C gilts had follicular cysts (P < .05). At 48 h after onset of estrus, plasma progesterone concentration tended to be greater (P = .13) for P gilts than for C gilts. In Exp. 2, gilts were ovariectomized 18 h after the onset of estrus. Ova were dissected from follicles on one ovary, and follicular fluid was aspirated from the other ovary. No differences were detected for the percentages of ova in various stages of meiosis. For APY gilts, follicular fluid estradiol concentration for P gilts tended to be lower (P = .12) than that for C gilts; however, no differences were detected for RSY gilts. Progesterone concentration in follicular fluid tended (P = .14) to be greater in P gilts than in C gilts. These results indicate that the effect of genetic line on age at puberty should be considered when inducing puberty.

Aging↗

A note on the effect of continuous ram presence on estrus onset, estrus duration and ovulation time in estrus synchronized ewes.

During fall season, 18 multiparous Corriedale ewes were divided into two equal groups for the continuous (CON) and intermittent (INT) presence of a ram. Estrus was synchronized with fluorgestone acetate intravaginal sponges that were left 14 days, plus an injection of 200&mgr;g of a prostaglandin F-2alpha analog at sponge removal. Estrus was detected three times a day (at 6 a.m., 2 p.m. and 10 p.m.) by using rams with harnessess and markers. Ovulation time was determined by laparoscopy, starting 24h after estrus detection. Estrus onset was (mean+/-S.E.M.) 32.9+/-1.6 and 45.3+/-4.4h for the CON and INT groups, respectively (P<0.01). Estrus duration was 31.1+/-0.9 and 30.2+/-1.2h, for the same groups, respectively (P>0.05). Ovulation time and the interval from sponge removal to ovulation (ISRO) for the CON and INT groups was 29.0+/-1.5, 62.0+/-2.0, 26.7+/-1.3 and 72.0+/-4.2h, respectively. Ovulation time was not different (P>0.05), but ISRO was shown to be different between treatments (P<0.05). It is concluded that the continuous presence of a ram after sponge removal hastens estrus onset and reduces the interval between sponge removal and ovulation, without modifying estrus duration and time between estrus onset and ovulation.

Journal Article↗

Activity of 3beta-hydroxyteroid dehydrogenase in the postovulatory follicle of the domestic fowl (Gallus domesticus).

The activity of delta5-3beta-hydroxysteroid dehydrogenase (3beta-HSD) in the postovulatory follicle (POF) of the domestic hen was studied; it declined rapidly during the first 15 h after ovulation and then fell gradually until 52 h when there was no detectable activity. Comparison of 3beta-HSD and glucose 6-phosphate dehydrogenase activities in the theca and granulosa cells indicated that the changes in enzymatic activity can be attributed to the thecal component; the enzyme activity in granulosa cells was stable up to 35 h after ovulation. The results appear consistent with the hypothesis that the POF is a source of steroid hormones.

Animals↗