Intraperitoneal insemination of the guinea pig with synchronized estrus induced by progesterone implant.
(1/818)
Female guinea pigs with synchronized ovulation by means of implantation of progesterone-filled tubing (P-tube) followed by a progesterone injection, were inseminated by intraperitoneal injection with sperm suspension. First, to obtain the optimum conditions for insemination, the females were inseminated singly over the range of 1-10 x 10(7) spermatozoa before and after the synchronized ovulation. The incidence of conception and implantation was 100% in the females given more than 5 x 10(7)/animal at 9:00 h on the 5th day after removal of the P-tube. Second, the reproductive ability of the inseminated females under this optimal condition was observed throughout the pregnancy to delivery. Inseminated females had a mean +/- S.D. gestation period of 68.7 +/- 0.5 days, a litter size of 2.8 +/- 0.6 pups and body weight of 110 +/- 14 g. These data were comparable to those of naturally-mated females. Our findings suggest that the artificial insemination by intraperitoneal injection in combination with the synchronized estrus technique is very useful for production control in a small colony of guinea pigs. (+info)
Characterization of uterine leukocyte infiltration in gilts after artificial insemination.
(2/818)
The objective of this study was to characterize the uterine leukocyte influx after artificial insemination (AI). After detection of oestrus with a boar at intervals of 1.5 h, seventy-two gilts were randomly assigned to a 2 x 3 x 4 factorial arrangement. AI was performed with 100 ml extended semen containing 5 x 10(9) spermatozoa (semen; n = 36) or 100 ml VSP semen extender (extender; n = 36) at one of three times after detection of oestrus: 12, 24 or 36 h (n = 24/time). The uterus was lavaged at 6, 12, 18 or 24 h (n = 18/time) after AI to determine the total number of uterine leukocytes. In addition, uterine lavage was performed on nine untreated gilts immediately after the detection of oestrus to establish a baseline number of leukocytes. The leukocyte response in all samples consisted predominately (92-99%) of polymorphonuclear neutrophilic granulocytes (PMNs). The mean number of PMNs recovered from the uteri of gilts treated with semen was greater than in gilts treated with extender and in untreated gilts (P < 0.01). The greatest number of PMNs in semen-treated gilts was found 12 h after AI (P < 0.01), and this number was sustained for 24 h. In contrast, the number of uterine PMNs recovered from extender-treated gilts reached a peak at 6 h and had declined by 12 h after AI (P < 0.05). It was concluded that an extensive influx of PMNs into the uterus is a normal sequence to AI. The consequences and importance of semen-induced uterine leukocytosis needs further investigation. (+info)
Sperm migration into and through the oviduct following artificial insemination at different stages of the estrous cycle in the rat.
(3/818)
In order to examine whether sperm migration into and through the oviduct follows an invariable pattern or is subject to regulation, rats in proestrus, estrus, metestrus, or diestrus were inseminated in the upper third of each uterine horn with 10-20 million epididymal spermatozoa. Three or eight hours later, the numbers of spermatozoa free and adhering to the epithelium in the ampullary and isthmic segments were determined. A significantly higher number of spermatozoa were recovered in estrus than in other stages, at 3 h than at 8 h, and at all stages from the isthmus than from the ampulla. Spermatozoa adhering to the epithelium were observed only in proestrus and estrus and in the isthmus. The effect of exogenous estradiol-17beta (E2) and progesterone (P4) on sperm migration was investigated in rats in which the estrous cycle was inhibited pharmacologically. E2 facilitated sperm migration into the oviduct and P4 antagonized this effect, whereas P4 alone had no effect. Concomitant treatment with E2+P4 induced adhesion of spermatozoa to the oviductal epithelium. In conclusion, the pattern of sperm migration into and through the rat oviduct varies with the stage of the cycle, being dependent on E2 and P4. The adhesion of spermatozoa to the rat oviductal epithelium is stage- and segment-specific and requires the combined action of both hormones. (+info)
Serum progesterone in predicting pregnancy outcome after assisted reproductive technology.
(4/818)
PURPOSE: Our purpose was to determine whether serum progesterone predicts pregnancy outcome after superovulation. METHODS: One hundred twenty-three consecutively pregnant patients were divided into three groups: group I, 55 patients following superovulation for assisted reproductive technologies; group II, 23 patients after correction of oligoovulation; and group III, 45 patients who conceived spontaneously. When beta-human chorionic gonadotropin was positive, progesterone was measured on the same serum sample. A serum progesterone level of 45 microns/L was set to differentiate between nonviable pregnancy and ongoing pregnancy. RESULTS: In group I, zero (0%) of 38 ongoing pregnancies and 10 (59%) of 17 nonviable pregnancies were observed with a progesterone level of < 45 microns/L [14.2 ng/ml (P < 0.001)]. In group II, 4 (27%) of 15 ongoing pregnancies and 5 (63%) of 8 nonviable pregnancies had a progesterone level of < 45 microns/L (P = NS). In group III, 10 (42%) of 24 ongoing pregnancies and 15 (71%) of 21 nonviable pregnancies were observed with a progesterone level of < 45 microns/L (14.2 ng/ml) (P = NS). CONCLUSIONS: A serum progesterone level of < 45 nM predicts nonviable pregnancy after superovulation for assisted reproductive technology. (+info)
Status of genomic imprinting in mouse spermatids.
(5/818)
The advent of human round spermatid microinjection (ROSI) into oocytes as a treatment for severe male infertility raises the question of whether spermatids have undergone all of the maturation processes necessary for normal development. It is particularly important to know whether spermatids have undergone correct genomic imprinting, which results in the parent-of-origin-specific expression of only one allele of a gene. We assessed the imprinting status of three maternally and three paternally expressed genes in interspecific hybrid embryos generated by injecting Mus castaneus spermatids into Mus musculus oocytes. We used the single nucleotide primer extension (SNuPE) assay to measure the relative expression of maternal and paternal alleles on the basis of sequence polymorphisms in the transcripts. Expression of imprinted genes in mouse embryos derived by ROSI did not differ from controls, indicating that paternal genes have undergone proper imprinting by the round spermatid stage. (+info)
Estrus synchronization of beef cattle with a combination of melengestrol acetate and an injection of progesterone and 17beta-estradiol.
(6/818)
Our hypothesis was that estrus synchronization in beef cattle using melengestrol acetate (MGA) and an injection of progesterone (P4) and 17beta-estradiol (E2) to regress dominant ovarian follicles would improve pregnancy rate (number conceived/number in group) to AI compared with feeding only MGA or injecting PGF2alpha. During 2 yr, peripubertal heifers (n = 52) and cows (n = 327) received either 1) MGA for 18 d (d 0 = 1st d of MGA) plus an injection of P4 and E2 in sesame oil (vehicle) on d 11 to regress persistent ovarian follicles (MGA+P4), 2) MGA for 18 d plus vehicle on d 11 (MGA), or 3) two injections of PGF2alpha 10 d apart (d 7 and 17, PG). Concentration of P4 was assessed in blood samples obtained on d 0, 7, and 17 to indicate estrual status (anestrual or estrual) during treatment to induce estrus synchrony. Observations for detection of estrus occurred every 6 h for 180 h following treatment cessation. Females showing estrus were inseminated 6 to 12 h after estrus detection. Conception to AI was determined by ultrasonography 35 to 40 d later. Conception rate was greater (P < .05) in females in the PG than in those in the MGA group but did not differ from conception rate of females in the MGA+P4 group. Among anestrual females, estrus synchrony rates were greatest (P < .10) among females treated with MGA+P4. Among females that were estrual before treatment cessation, estrus synchrony rates were greater (P < .10) among females treated with MGA+P4 or PG than among those given MGA. Pregnancy rates were greater (P < .05) among females that were anestrual before treatment cessation and treated with MGA or MGA+P4 than among those treated with PG. Estrus synchronization using MGA+P4 and E2 differentially improves estrus synchronization and pregnancy rates among anestrual and estrual beef cattle while maintaining conception rates similar to those of PGF2alpha-treated females. (+info)
Mouse spermatid nuclei can support full term development after premature chromosome condensation within mature oocytes.
(7/818)
The nucleus of round spermatids, the earliest haploid male germ cells, can participate in the formation of normal zygotes when incorporated into activated oocytes. In this study, we injected mouse round spermatids into homologous mature oocytes that were kept arrested at metaphase II to induce premature chromosome condensation (PCC) of the spermatid nuclei. After full condensation of the spermatid chromosomes, the oocytes were activated by Sr2+-containing medium, into which cytochalasin B was added to prevent extrusion of the segregated female and male chromosomes as polar bodies. Out of 142 oocytes examined, 104 (73%) formed two male (pseudo)pronuclei and two female pronuclei. To restore the diploid state of these zygotes, one of the female pronuclei was removed. When cultured in vitro for 72 hours, all (n = 37) of the constructed embryos developed to the morula/blastocyst stage. When 2-cell embryos and morulae/blastocysts were transferred into pseudopregnant females, 14 (13/96) and 24% (9/37), respectively, developed into term offspring. This study indicates that the spermatid chromosomes, which had undergone PCC, moved safely to opposite poles after oocyte activation. Since round spermatids contain no (in the mouse) or little (in patients with spermatogenic failure) oocyte-activating factor, this method may be used to rescue oocytes that fail to be activated at the time of spermatid injection. (+info)
Duration of estrus in relation to reproduction results in pigs on commercial farms.
(8/818)
This research was conducted to determine factors that influence duration of estrus, AI strategy, and reproduction results between and within commercial swine farms that use AI. Data from 15,186 sows and gilts on 55 farms for a period of 6.1+/-4.2 mo per farm were used in this study. The average duration of estrus was 48.4+/-1.0 h, ranging from 31 to 64 h, and was consistent from month to month within a farm (repeatability of 86%). Differences in duration of estrus between farms accounted for 23% of the total variation in duration of estrus. On most farms (n = 45), gilts showed a shorter (P < .05) duration of estrus than sows (40.8+/-1.1 h vs 48.5+/-1.0 h). The duration of first estrus after weaning was longer (P < .0001) compared with that of repeat-breeder sows (50.2+/-1.0 h vs 46.8+/-1.0 h). Duration of estrus decreased (P < .05) when interval from weaning to estrus increased from 4 to 6 d (56.0 +/- 1.2 h vs 45.8 +/-1.2 h). The regression of interval from onset to estrus to first AI and interval from weaning to estrus varied between farms and ranged from -7.4 to +1.3 h/d; four farms had a positive relationship. Farrowing rate decreased (P < .05) from 89.7+/-2.7% to 78.2+/-5.74 when the interval from weaning to estrus increased from 4 to 10 d. The litter size decreased (P < .05) from 11.7 to 10.6 pigs when the interval from weaning to estrus increased from 4 to 7 d. Compared with a single AI, double AI in sows and gilts resulted in a 4.3 and 7.0% higher (P < .05) farrowing rate, respectively. When the first AI was performed after expected ovulation, reproduction results were lower than when AI was performed before or at expected ovulation in sows. Duration of estrus was not related to farrowing rate or litter size in individual pigs. Number of inseminations per estrus, time of AI, and duration of estrus were correlated, which made it difficult to assess which of these factors was primarily related to the farrowing rate or litter size. Knowledge of average duration of estrus on farms and of factors that influence the duration of estrus on commercial farms can help to improve the efficiency of the AI strategy specific for each farm. (+info)