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1.
Early embryonic development and implantation were studied in tropical short-nosed fruit bat Cyanopterus sphinx. We report preimplantation development and embryo implantation. Different stages of cleavage were observed in embryo by direct microscopic examination of fresh embryos after retrieving them either from the oviduct or the uterus at different days, up to the day of implantation. Generally, the embryos enter the uterus at the 8-cell stage. Embryonic development continued without any delay and blastocyst were formed showing attachment to the uterine epithelium at the mesometrial side of the uterus. A distinct blue band was formed in the uterus. The site of blastocyst attachment was visualized as a blue band following intravenous injection of pontamine blue. Implantation occurred 9+/-0.7 days after mating. This study reports that bat embryonic development can be studied like other laboratory animals and that this bat shows blue dye reaction, indicating the site and exact time of implantation. This blue dye reaction can be used to accurately find post-implantational delay. We prove conclusively that this species of tropical bat does not have any type of embryonic diapause.  相似文献   

2.
The western spotted skunk exhibits an obligate delay of implantation lasting 200-220 days. The pituitary is essential for luteal activation. The corpora lutea, in turn, secrete the hormones necessary for blastocyst implantation. Two experiments were designed to determine which pituitary hormones are responsible for increasing luteal activity and induction of implantation. Forty-two pregnant skunks with delayed implanting blastocysts were treated as follows: 13 served as untreated controls, 6 received 0.5 mg prolactin (PRL) daily, and 5 received diluent beginning in January. Four received 1.5 mg bromocriptine (CB-154) daily, 3 received both CB-154 and PRL, 3 received diluent, 5 received a gonadotropin-releasing hormone agonist (GnRHa) dispensed from osmotic minipumps, and 3 received diluent dispensed from osmotic minipumps starting in April. The skunks were subjected to a natural photoperiod. Duration of preimplantation and blood levels of progesterone and luteinizing hormone were measured. PRL significantly (p less than 0.05) shortened and CB-154 significantly (p less than 0.05) prolonged the duration of preimplantation when compared to controls (148 +/- 33.6 vs. 251 +/- 3.2 vs. 199 +/- 5.1 days, respectively). PRL was able to reverse the inhibitory effect of CB-154 when both were administered simultaneously (195 +/- 4.0 vs. 251 +/- 3.2 days). GnRHa had no significant (p greater than 0.05) effect on duration of preimplantation (199 +/- 5.1 days) when compared to controls (203 +/- 3.2 days). These results indicate that PRL is the primary pituitary hormone responsible for increased luteal activity and subsequent blastocyst implantation in the spotted skunk.  相似文献   

3.
The objective of this study was to evaluate if treatment of eCG-superovulated goats with fluorogestone acetate (FGA) would increase the number and quality of embryos recovered. Goats (n = 25) were given an intravaginal sponge containing 45 mg FGA for 12 days, with 1000 IU eCG and 7.5mg of Luprostiol (a PGF(2 alpha) analog) given -48 and 0 h relative to sponge removal. Goats were mated by natural service every 12h during estrus and surgical embryo collection was done 6 days after the last mating. There were two treatment groups; those in the FGA group (n = 13) had a FGA sponge from 8h after mating to embryo collection, whereas goats in the control group (n = 12) did not receive any post-mating treatment. Premature luteal regression occurred in 61.5% (8/13) and 83.3% (10/12) of the goats in the FGA and the control groups, respectively (P > 0.05). Corpus luteum life span averaged 4 days in goats with premature luteolysis. The mean (+/- S.E.) number of transferable embryos was 5.7 +/- 1.6 in the FGA group and 0.1 +/- 0.1 in the control group (P < 0.05). Within the FGA group, the embryo recovery rate was similar in goats with premature luteal regression compared to those with normal luteal function, although non-transferable embryos were only found in goats with premature luteal regression. In conclusion, post-breeding treatment with FGA increased embryonic survival in eCG-superovulated goats, even though it did not prevent premature luteal regression.  相似文献   

4.
Surgical transfer of in vivo produced conventionally frozen-thawed embryos of farmed European polecat (Mustela putorius) was investigated as a part of an ex-situ preservation program which has the long-term aim of developing a genome resource bank for the endangered European mink (Mustela lutreola). Eighteen oestrous yearling European polecat donors were mated once daily on two consecutive days using 13 fertile males. The donors were surgically flushed for embryos 8-9 days after the first mating. The embryo recovery rate was 60% (116 embryos/193 corpora lutea). The embryos were cryopreserved with 1.5 M ethylene glycol in a programmable freezer using a conventional slow freezing protocol. The thawed embryos were surgically transferred either after dilution with 0.5 M sucrose or directly without removal of ethylene glycol. To induce ovulation, eight recipient females were mated once daily on two consecutive days with vasectomized males starting 7 or 8 days before embryo transfer. The recipients received 7-11 embryos each and three recipients delivered a total of nine pups after a gestation length of 44-46 days. The embryo survival rate was 10% (9 pups/93 frozen embryos). This report describes the first successful cryopreservation of embryos in the Mustelidae family resulting in viable offspring. The low embryo survival rate, however, indicates that the freezing-thawing protocol needs to be improved.  相似文献   

5.
Mink are seasonal breeders that display an obligate delay preceding implantation and a post implantation gestation of 31 d. The purpose of this study was to evaluate gestational parameters in mink by ultrasonography. A total of 92 female mink were mated twice during the period from March 2 to 20. The mink were scanned once and allowed to whelp (n=55); or scanned at 3 to 5-d intervals until parturition (n=13); or immediately subjected to autopsy (n=24) after scanning. Embryonic age was calculated from the date of parturition or from crown rump length. Uterine swelling diameter and fetal head size were correlated with embryonic age. The gestational sac grew rapidly once implantation had occurred. Uterine swellings of 4 to 5 mm in diameter were found on Days 2 to 4 post implantation and increased through Days 18 to 20, at which time they began to elongate due to the longitudinal growth of the fetus. Fetal cardiac activity could be detected on Days 10 to 12 post implantation in live embryos. The heart frequency was 198 +/- 3.0 beats per minute and did not vary from Days 12 to 30 post implantation. Fetal head diameter of 5 mm was first detected on Day 19 post implantation and grew gradually to 9 to 10 mm at parturition. It was not possible to accurately assess the number of conceptuses in utero. We conclude that ultrasonography can be employed in mink to diagnose pregnancy, to predict the parturition date and to determine the presence of live fetuses.  相似文献   

6.
The obligate embryonic diapause that characterizes gestation in mink engenders a developmental arrest at the blastocyst stage. The characteristics of escape from obligate diapause were investigated in embryos reactivated by treatment of the dams with exogenous prolactin. Protein and DNA synthesis showed marked increases within 72 h after the reinitiation of development, and embryo diameter increased thereafter. Trophoblast cells from embryos at Day 5 after activation proliferated more readily in vitro than trophoblasts from diapause or from Day 9 after activation, while in Day 9 embryos, cells from the inner cell mass (ICM) replicated comparatively more readily in vitro. There was evidence of expression of fibroblast growth factor-4 (FGF4) in both diapause and activated embryos and in ICM, but not the trophoblast. FGF receptor-2 was present in embryos from Day 5 after reactivation in both trophoblast and ICM cell lines. Trophoblast cell lines established from mink embryos proliferated in culture in the presence of FGF4 with a doubling time of 1.4 days, while in its absence, the doubling time was 4.0 days. We conclude that, during reinitiation of embryogenesis in the mink after diapause, embryo growth is characterized by gradual increases in protein synthesis, accompanied by mitosis of the trophoblast and ICM. There appears to be a pattern of differential proliferation between cells derived from these embryonic compartments, with the trophoblast phase of replication occurring mainly in the early reactivation phase, while the ICM proliferates more rapidly nearer to the time of implantation.  相似文献   

7.
Surgical embryo transfer of farmed European polecat (Mustela putorius) was investigated as part of an ex situ preservation project. The long-term objective of the project is to develop effective technology for ex situ conservation of the European mink (Mustela lutreola), which is a highly endangered aboriginal European species. Twenty European polecat females, which served as a model species for the European mink, were humanely killed 4-9 days after first mating and embryos were recovered from oviducts and uteri. Donor-recipient pairs (n = 16) were generated by mating the donors (n = 20) once a day for 2 consecutive days with fertile males and by mating the corresponding recipients (n = 16) on the same days with vasectomized males. An embryo recovery rate of 70% (200 recovered embryos/284 corpora lutea) was achieved from 20 donors. Morulae and blastocysts were recovered between Days 5 and 9 after first mating and were regarded as the best developmental stages for uterine embryo transfer. A total of 172 embryos were transferred surgically under general anaesthesia into the ovarian third of the left uterine horn of 16 recipients with a thin glass capillary. Eleven recipients (69%) produced 72 pups equivalent to an average success rate of 42% (72 pups/172 transferred embryos). The average litter size was 4.5 (range 0-9). These results with this model species, farmed European polecat, demonstrate the potential of embryo transfer as an effective method for the preservation of the endangered European mink (M. lutreola). These species are closely related and have a similar reproductive physiology. However, success of applying embryo transfer in conserving European mink is still dependent on further studies both into its reproductive physiology and developing of improved flushing techniques for anaesthetized donors and the successful transfer of frozen-thawed embryos.  相似文献   

8.
Primiparous crossbred sows (n = 43), lactating for an average of 21.1 +/- 0.1 d and weaning 8.7 +/- 0.1 pigs, were used to evaluate the influence of insulin on ovulation rate and embryo survival. The sows were maintained on 2.3 kg/head/d of a 14% protein gestation diet during pregnancy, fed ad libitum during lactation, given 2.7 kg/head/d from weaning until re-breeding and fed 2.3 kg/head/d after mating. Beginning the day after weaning (Day 0) sows were treated with 0.4 IU/kg body weight (BW) insulin (n = 21) or were administered an equivalent volume of saline (n = 22) for 4 d. Beginning on Day 3 and continuing until Day 14 after weaning, the sows were checked for estrus twice daily and were artificially inseminated using pooled semen from 2 fertile boars. At slaughter (days 30 to 40 of gestation), ovaries and uteri were collected, and the ovulation rate, embryo number and viability, and uterine weight and length were evaluated and recorded. Use of insulin decreased the average interval from weaning to estrus compared with saline by increasing percentage in estrus by Day 14 after weaning (5.0 +/- 0.57 vs 6.9 +/- 0.56 d, respectively; P < 0.03). Ovulation rate, number of embryos, embryo survival, and average uterine length and weight were not influenced by insulin treatment. Overall, insulin affected reproductive efficiency in primiparous sows by increasing the percentage of sows in estrus.  相似文献   

9.
We have developed a technique to sample the preimplantation embryo, which may, in the future, be applied to prenatal diagnosis of genetic disease. Using micromanipulation, we aspirated a single blastomere from 4-cell mouse embryos. This procedure had no effect on in vitro development; 98% of control and 94% of biopsied embryos reached the blastocyst stage after 48 h in culture. Furthermore, after transfer to pseudopregnant recipient mice, the rate of fetal development of biopsied embryos was not significantly different from control embryos, although implantation rate was significantly reduced (mean +/- SD: biopsied 53.1 +/- 4.0, control 81.8 +/- 8.4, p less than 0.001). For the first time we have produced monolayer cell cultures derived from single preimplantation blastomeres. Individual biopsied blastomeres were cultured in vitro on different extracellular matrix components. Significantly greater cell proliferation was obtained in wells coated with fibronectin (FN), laminin (LN), and a complex of laminin and nidogen (LNC) than in a less specific matrix of swine skin gelatin (SSG). Mean (+/- SE) cell nuclei number per well after 6 days in culture was 6.4 +/- 2.1, 11.9 +/- 1.5, 19.8 +/- 2.9, and 20.9 +/- 2.6 in wells coated with SSG, LN, FN, and LNC respectively.  相似文献   

10.
Basigin is a member of the immunoglobulin superfamily and a key molecule related to mouse blastocyst implantation. Whether preimplantation mouse embryos express basigin mRNA is still unknown. The aim of this study was to use a quantitative competitive polymerase chain reaction to assess quantitatively the levels of basigin mRNA in mouse oocyte and preimplantation embryos. Basigin mRNA was detected in the oocyte and all the stages of preimplantation embryos. The levels of basigin mRNA were 0.0606 +/- 0.0282 in the oocyte, 0.0102 +/- 0.0036 in the zygote, 0.0007 +/- 0.0003 in the 2-cell embryo, 0.0031 +/- 0.0017 in the 4-cell embryo, 0.0084 +/- 0.0024 in the 8-cell embryo, 0.0537 +/- 0.0121 in the morula and 0.0392 +/- 0.0161 attomoles in the blastocyst, respectively. The levels of basigin mRNA in the oocyte, morula and blastocyst were significantly higher than those in the zygote and embryos at the 2-cell, 4-cell and 8-cell stages. The high level of basigin expression in the blastocyst may play a role during embryo implantation.  相似文献   

11.
In 1980 and 1981, respectively, 12 and 19 one-year-old mink females of standard and jetblack breeds were used to determine the progesterone level approximately at the time of expected implantation. Blood samples were collected every 2 to 3 days or daily in order to accurately estimate the time of increase in plasma progesterone levels. The results indicate that the progesterone level increased to above 10 ng/ml, 31.6 (sigma = 1.3) days prior to parturition on the average. This was supported by physiological explanations. The date of mating in mink had an effect on the date of increase in plasma progesterone. Matings at a late date in the estrous period, reduced the period of delay before implantation. Nevertheless, both the dates of implantation and parturition were delayed compared with the results of earlier matings in mink.  相似文献   

12.
Early embryonic development and in vitro culture of in vivo produced embryos in the farmed European polecat (Mustela putorius) was investigated as a part of an ex situ conservation program of the endangered European mink (Mustela lutreola), using the European polecat as a model species. The oestrus cycles of 34 yearling polecat females were monitored by visual examination of the vulval swelling and, to induce ovulation, the females were mated once daily on two consecutive days. Sixteen yearling males were used for mating. The females were humanely killed 3-14 days after the first mating and the uteri and oviducts were collected for embryo recovery. Uterine and oviductal flushings yielded a total number of 295 embryos, representing developmental stages from the 1-cell stage to large expanded and hatched blastocysts. On Day 3 after the first mating, only 1-16-cell stage embryos were recovered. Between Days 4 and 6 after the first mating, 1-16-cell stage embryos and morulae were found. The first blastocysts were recovered on Day 7 after the first mating. The first implanted blastocysts were detected on Day 11 after the first mating. A total number of 85 embryos were in vitro cultured after recovery. Blastocyst production rates for in vitro cultured 1-16-cell stage embryos and for morulae/compact morulae were 68 and 84%, respectively. For all cultured embryos, the hatching rate was 15%. The in vitro culture requirements for the preimplantation embryos of the farmed European polecat remain to be determined before further utilization of the technique.  相似文献   

13.
Length of gestation was studied in pregnancies established by intrabreed, interbreed, and mixed-breed embryo transfers of Finnish Landrace (Finn; mean gestation: 144.9 days) and Targhee (mean gestation: 150.4 days) embryos to Finn cross and Targhee recipients. At least one lamb of each breed comprised mixed-breed pregnancies. There was a significant effect of the breed of lamb (Finn, Targhee, or Finn and Targhee) on the length of gestation (P < .01), but not of breed of recipient. Mixed-breed pregnancies had a mean gestation period intermediate between those for pregnancies which contained only one breed of lamb. The mean gestation period for mixed-breed pregnancies was 2.10 +/- .70 (x +/- S.E. ) days longer than for pregnancies with only Finn lambs (P < .01) and 2.99 +/- .73 days shorter than for pregnancies with only Targhee lambs (P < .001). The delay in parturition in mixed-breed pregnancies beyond the normal gestation period for Finn lambs occurred even in litters with a majority of Finn lambs. These results demonstrate an interaction between fetuses in the processes leading to parturition. Possible mechanisms by which the Targhee lamb delayed parturition in mixed-breed pregnancies are discussed.  相似文献   

14.
The effects of day of mating and litter size on gestation length in dogs were studied in 36 beagle bitches (age 2-10 yr). The day that plasma progesterone concentrations exceeded 2 ng/mL was considered the day of ovulation; dogs were randomly assigned to be bred once, 1-5 days after the estimated day of ovulation. The interval from mating to parturition was negatively correlated with the number of days from estimated ovulation to mating (P < 0.01). Gestation length (interval from ovulation to parturition) was almost constant at 63.9 +/- 0.2 days (mean +/- S.E.M.), with no significant relationship between the number of fetuses and the duration of gestation.  相似文献   

15.
Mink ovariectomized 14 days after the first of two matings received injections of 2 mg MPA, the same MPA treatment + an oestradiol-17 beta implant or no replacement therapy. Some mink were ovariectomized after implantation and given a single dose of 2 mg MPA or no replacement therapy. MPA persisted in the serum at detectable levels for 13 or more days in all mink treated. MPA and MPA + oestradiol induced uterine growth but neither treatment was capable of inducing embryo implantation. More embryos were retained in mink treated with MPA alone and these appeared to be viable. Implanted embryos persisted for a longer period in animals that were ovariectomized and treated with MPA. MPA neither supported pregnancy nor permitted parturition. Serum LH was elevated by 1 week after ovariectomy and elevations persisted for a further 20 or more days. While MPA alone had no apparent negative feedback effects on LH, animals that received MPA + oestradiol did not display any elevation of LH, suggesting that oestradiol or a combination of MPA and oestradiol has a potent negative feedback in mink.  相似文献   

16.
The objective of these studies was to determine the effect of progesterone and medroxyprogesterone injected at varying times post coitum on gestation length and litter size. During a 3-year period mink females of "Standard" strain were given progesterone at a dose 5 mg at 15 and 20 days in the first year and at 17-20 days in the second year, after the last mating. The respective control groups were given the vehicle. Medroxyprogesterone acetate was given in the second year at a dose of 4 mg at 14-19 days following the last mating and in the third year at a dose of 2 mg at 8 days after the last mating. The results on pregnancy length and litter size after progesterone injections in the experimental and respective control groups were as follows: 52.7 days and 4.3 kits, 51.3 days and 4.3 kits, 52.2 days and 5.8 kits, 50.2 days and 4.8 kits, 44.7 days and 5.6 kits, 46.0 days and 6.1 kits. A dose of 4 mg MPA resulted in the blockage of parturition in pregnant females. After administration of MPA at doses of 2 mg at 8 days following the last mating, the pregnancy length and litter size in the experimental group were: 48.0 days and 6.1 kits and in the control: 52.2 days and 4.8 kits.  相似文献   

17.
The energy metabolism of preimplantation embryos can be used to predict viability and postimplantation development. Although preimplantation development and mean blastocyst cell numbers of goat in vitro-fertilized (IVF) embryos and chemically activated parthenogenotes are comparable, mammalian parthenogenotes are not viable, with most dying shortly after implantation. The objective of this study was to compare glucose and pyruvate metabolism of IVF goat blastocysts with that of parthenogenetic blastocysts developing from chemically activated oocytes. Embryos derived from IVF and parthenogenotes produced by exposing oocytes to either ionomycin or ethanol followed by 6-dimethylaminopurine (6-DMAP) were cultured in G1.2/G2.2 sequential culture media. Metabolism was determined for individual blastocysts using [5-3H]glucose and [2-14C]pyruvate to determine glycolytic and Kreb's cycle activity, respectively. Data were analyzed by ANOVA. A significantly higher percentage of activated oocytes underwent cleavage and developed to the blastocyst stage compared to IVF oocytes (p < 0.05). There was no significant difference in glucose or pyruvate metabolism between IVF and parthenogenetically activated blastocysts. Mean glucose metabolism through glycolysis was 154.9 +/- 29.1, 130.3 +/- 17.1, and 129 +/- 16.5 pmol/embryo/3 h for IVF, ethanol-activated, and ionomycin-activated blastocysts, respectively. Mean pyruvate metabolism through the Kreb's cycle was 28.1 +/- 8.0, 15.8 +/- 4.2, and 24.4 +/- 4.4 in pmol/embryo/3 h for IVF, ethanol-activated, and ionomycin-activated blastocysts, respectively. Our results suggest that known differences in postimplantation development observed in IVF versus parthenogenetic embryos cannot be attributed to differences in pyruvate or glucose metabolism in the preimplantation blastocysts. Thus, these activation protocols result in embryos capable of appropriate regulation of key metabolic enzymes.  相似文献   

18.
19.
BACKGROUND: Lasofoxifene is a nonsteroidal selective estrogen receptor modulator (SERM). With high affinity to the alpha and beta human estrogen receptors and greater potency than other SERMs, lasofoxifene is potentially a superior treatment for postmenopausal osteoporosis. In light of the known effects of estrogen-modulating compounds on female reproductive indices, two studies were conducted to evaluate the effects of lasofoxifene on female rat cyclicity, reproduction, and parturition. METHODS: One study evaluated effects of lasofoxifene on estrous cyclicity, and the second study assessed effects on implantation and parturition. In the cyclicity study, lasofoxifene was administered to female rats at doses of 0.1, 0.3, and 1.0 mg/kg/day for 14 consecutive days. After treatment, there was a 3-week reversibility phase followed by a mating phase. In the implantation study, lasofoxifene was administered to pregnant female rats at doses of 0.01, 0.03, and 0.1 mg/kg/day for 7 consecutive days (gestation day [GD] 0-6). Some animals were euthanized on GD 21, and the remainder of the group was allowed to deliver the F1 generation. Several developmental indices were evaluated in the F1 pups through post-natal day (PND) 21. RESULTS: In the cyclicity study, all lasofoxifene-treated females were anestrous by Study Day 7 (1.0 mg/kg) or 9 (0.3 and 0.1 mg/kg). The reversibility phase resulted in restoration of normal estrous cycles by the end of 1 (0.1 mg/kg) or 2 weeks (0.3 and 1.0 mg/kg). During the mating phase, no adverse effects occurred in pregnancy success or reproductive parameters. In the implantation study, all doses of lasofoxifene increased pre- and post-implantation losses, increased gestation length, and reduced litter size. None of the developmental parameters measured on the F1 generation was adversely affected. CONCLUSION: Lasofoxifene reversibly altered the estrous cycle and inhibited implantation, consistent with what would be expected from a member of the SERM class.  相似文献   

20.
The length of gestation in Andalusian, or Spanish Purebred (SPB) and Arabian (AB) mares reared in Spain was analysed, based on 766 spontaneous full-term deliveries appertaining to 141 mares of SPB breed and 72 mares of AB breed in 31 breeding seasons. The data were obtained from the Yeguada Militar de Jerez de la Frontera stud farm in Cádiz, Spain. The mean length of gestation was of 336.8+/-0.48 days in the SPB mares and 340.3+/-0.63 days in AB mares. To assess the accurate prediction of time of birth the potential effect of a number of factors was investigated. The influences of the breed, mare, month and year of mating, age of the mother, number of births and sex of the foal were statistically significant. The factor have the greatest influence over the gestation length was the mare itself, with a correlation among consecutive births of around 0.4. The effect of inbreeding, both of the mare and foal, was negligible. Gestation length shortened as the breeding season progressed: in both breeds, a delay of 1 month in mating corresponded to a decrease of 3 days in the gestation length. According to our results, gestation length decrease as the mare gets older, with the shortest gestation periods when the mare is 10-12 years old, and from this point on, it slowly increases. The gestation period shortens as the 4th or 5th birth approaches, and then gets progressively longer. The range of variation in gestation length due to the number of births to the mare is of 2.9 days for the AB mares, and 2.2 days for SPB mares. The heritability for the gestation length for AB and the SPB breeds was 0.2, with a repeatability of 0.36 and 0.37, for SPB and AB breeds, respectively. With the data from both breeds, and using a classical approach, the response to selection was estimated if mares with extreme gestation lengths were culled, i.e. lengths which are under 310 days, or over 360 days. According to our results, in the case of SPB, a decrease of 14-45% would occur in the number of extreme gestation lengths, while in the AB breed, this value would decrease from 2 to 39%.  相似文献   

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