摘要:

AbstractMature female Chinese hamsters ovulate an average of 8.8 ± 1.0 (mean ± SD) eggs per female in each estrous cycle. Superovulation can be induced in both immature and mature females by subcutaneous or intraperitoneal injections of pregnant mare serum gonadotropin (PMSG) and either human chorionic gonadotropin (hCG) or pituitary luteinizing hormone (PLH). The best superovulation in immature females was induced by the administration of 15 IU of PMSG followed 72 hr later by injection of 15 IU of hCG (about 25 eggs per female) or 0.2 mg (200 IU) PLH (about 46 eggs per female). Ovulation started about 13–15 hr after administration of hCG (or PLH) and was completed during the next 5–6 hr. Superovulation in mature females could be induced by injecting PMSG any day of the estrous cycle, but the best superovulation (about 39 eggs per female) was induced by injecting 15 IU of PMSG on day 1 (day of ovulation) followed by the injection of 0.4 mg of PLH 72 hr later. When immature females treated with the best superovulatory protocol were mated on the evening of PLH injection, only 5% of the eggs were found fertilized 50 hr after PLH administration. On the other hand, about 60% of the eggs were found fertilized in mature females mated following treatment with the best superovulatory protocol. The majority (83–85%) of superovulated eggs obtained from both immature and mature females were normally fertilized i

ISSN:0148-7280    

DOI:10.1002/mrd.1120160402

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY

摘要:

AbstractChanges in intracellular localization of argyrophilic proteins visualized as silver‐stained particles by nuclear organizer region (NOR)‐silver staining were investigated in starfish oocyte maturation. The silver‐stained particles were localized in the germinal vesicle and nucleolus of immature oocytes and dispersed into the cytoplasm at the time of germinal vesicle breakdown (GVBD). In the mature egg cytoplasm, silver‐stained particles were distributed on yolk‐like granules with diameters of 0.3–1.0 μm. In spermatozoa, silver‐stained particles were detected heavily in the acrosome and centrosomes but few were detected in the nucleus, whereas they were present in the male pronucleus of fertilized eggs. The silver‐stained particles were removed by pretreatment of eggs with protease but not with nuclease. These results indicate that argyrophilic proteins disperse to the egg cytoplasm during GVBD and might be incorporated to the male pronucleus from the egg cytoplasm in fertilization. The morphological changes from chromosomes through chromosome vesicles to female pronucleus were also observed with light microscopy after NOR

ISSN:0148-7280    

DOI:10.1002/mrd.1120160403

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY

摘要:

AbstractA procedure was developed for isolation of plasma membrane (PM) preparations from unfertilized mouse eggs. Zona‐free mouse eggs prepared by the method of Boldt and Wolf (Gamete Res 13:213–222, 1986) were labeled with125I‐concanavalin A (ConA) prior to sonication and fractionation on iso‐osmotic self‐generated Percoll density gradients. Experiments using the ConA‐specific sugar α‐methylmannoside (αMM) indicated that125I‐ConA bound specifically to the egg PM. Greater than 95% of125I‐ConA binding to zona‐free eggs was blocked in the presence of 0.1 M αMM, and incubation of eggs in αMM after125I‐ConA labeling caused release of 85–90% of bound label. Fractionation of125I‐ConA‐labeled eggs by Percoll density gradient centrifugation yielded a single radioactive peak at density = 1.025, corresponding to egg PM material. Prolonged incubation of125I‐ConA‐labeled eggs or egg sonicates prior to fractionation did not alter the location of the radioactive peak, indicating that125I‐ConA did not label other organelles. As a control, human erythrocytes were labeled with125I‐ConA and fractionated under identical experimental conditions and yielded a single radioactive peak at density (1.020) comparable to that observed for125I‐ConA‐labeled eggs. These results indicate that125I‐ConA can be used as a specific marker to support PM isolati

ISSN:0148-7280    

DOI:10.1002/mrd.1120160404

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY

摘要:

AbstractAntisperm antibodies are implicated as one causative factor of infertility, but the target antigens have not been identified. Immune responses to sperm antigens are qualitatively variable even within a single mouse strain. We took advantage of this variability and immunized individual female mice to allogeneic sperm to reflect their natural exposure during mating. We determined the ability of the individual sera to inhibit in vitro fertilization and to bind to sperm antigens separated by electrophoresis. Compared to preimmune sera, four of five immune sera significantly inhibited in vitro fertilization. The serum from individual mice bound variable panels of sperm antigens. By comparing the panels, we identified two polypeptides with molecular weights of 40,000 and 44,000 that were bound by all sera. We propose that these molecules may be good candidates for further investigation of the immunoprophylaxis of pregnancy.

ISSN:0148-7280    

DOI:10.1002/mrd.1120160405

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY

摘要:

AbstractThe sex ratio of the fetuses from mice treated with pregnant mare's serum (PMS)/human chorionic gonadotropin (HCG) to induce ovulation did not differ appreciably from that of spontaneously ovulated controls. Rather, an intriguing observation was that the sex ratio in the right uterine horns tended to be lower than that in the left horns in both spontaneously ovulated controls and PMS/HCG‐treated groups. We speculate on its possible relation to the observed right‐left asymmetry of horn sizes (number of fetuses in the ho

ISSN:0148-7280    

DOI:10.1002/mrd.1120160406

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY

摘要:

AbstractZonae pellucidae (ZP) were isolated from 1,500 porcine ovaries and heat solubilized to generate approximately 15 mg ZP glycoprotein. Analysis of this material by isoelectric focusing, one‐dimensional electrophoresis, and gas chromatography indicated the presence of a major glycoprotein species that exhibited considerable microheterogeneity with respect to its charge (pI 7.5–3.5) and molecular mass (45–85 kDa) and that contained 39.6% carbohydrate, predominantly N‐acetylglucosamine.Chemical deglycosylation of porcine ZP using trifluoromethanesulphonic acid (TFMS) resulted in the production of five discrete protein bands on one‐dimensional sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS/PAGE) with molecular masses of 66, 52, 36, 32, and 16 kDa.Antisera raised in rabbits and marmosets to ZP and/or deglycosylated ZP (DGZP) were used in immunoblotting experiments to demonstrate the retention of immunogenicity by DGZP and the cross‐reactivity of the antisera with their heterologous antigen. These studies indicated that antisera that were capable of inhibiting the fertility of primates in vivo and the penetration of the human ZP in vitro reacted preferentially with 3 of the 5 products of deglycosylation, with molecular masses of 66, 52, and 36 kDa. Anti‐DGZP antibodies were also shown to interact with intact porcine and human ZP and, with the latter, to block the ability of human spermatozoa to both bind to and penetrate t

ISSN:0148-7280    

DOI:10.1002/mrd.1120160407

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY

摘要:

AbstractHuman oocytes were frozen and thawed by four methods previously used for cryopreser‐vation of human embryos. Most of these oocytes were inseminated after thawing to assess their capacity to fertilize and form pronuclear ova. Their morphology was assessed by phase‐contrast microscopy used in routine IVF. Twenty‐three oocytes were examined by electron microscopy to critically evaluate the effects of cooling and cryopreservation and to confirm fertilization.Morphological survival was observed in more than 60% of the oocytes examined after freeze‐thawing. The main features of cryoinjury were cracks in the zona pellucida, disruption of the plasma membrane and extensive disorganization of the ooplasm. Subtle changes in the cytosol of cumulus cells was also observed. Cooling to 0°C or −6°C had little effect on cytoplasmic structure. Spindles were damaged in two frozen oocytes.Cumulus cell activity, sperm binding to the zona, sperm penetration of the zona seem to be largely unaffected by freeze‐thawing. Fertilization was observed in eight oocytes after postthaw insemination and three embryos (8‐cell to morula stages) were developed from pronuclear ova on further culture. Both monospermic and polyspermic fertilization were confirmed by electron microscopy and micronuclei were detected in three pronuclear ova. The genetic implications of these nuclear aberrations are discussed.These preliminary studies indicate that oocyte freezing needs to be integrated cautiously with clinical IVF by further assessment of embryos developed from

ISSN:0148-7280    

DOI:10.1002/mrd.1120160408

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY

摘要:

AbstractEmbryos ofSminthopsis crassicaudataandSminthopsis macrourawere cultured for up to 96 hours during cleavage and early expansion of the blastocyst in Dulbecco's modified Eagle's medium (DMEG), DMEG containing 2.76 gm/liter sodium lactate (DMEGL), DMEG containing 3.5 gm/liter galactose (DMEGAL), DMEG containing 15 ng/ml progesterone (DMEGP) or 150 ng/ml progesterone (DMEGP10), and DMEGL containing 15 ng/ml progesterone (DMEGLP).The disappearance of sperm was used to indicate the time of ovulation (day 0). Fertilized eggs were found in the uterus at the end of day 1, four‐cell stages at the end of day 2, and embryos completing the fourth division by the end of day 3 inS. macrouraand day 4 inS. crassicaudata.Estimated developmental times in culture were similar to those obtained in vivo. In both species, the first two divisions take about 24 hours, cleavage is arrested for 24 hours or longer at the rounded four‐cell stage, and the third and fourth divisions take a further 24 hours. The blastocyst expands during the next 24 hours in which time the fifth and sixth divisions occur. It was possible to culture embryos fromS. macrourabut notS. crassicaudataover the four‐cell stage to early expanding blastocysts.DMEGAL did not support cleavage in culture. DMEG, DMEGL, DMEGP, DMEGP10, and DMEGLP all supported culture during cleavage and early blastocyst expansion. Blastocyst expansion was slightly enhanced using media containing sodium lactate. More embryos completed the fifth division and formed expanding blastocysts in DMEG, DMEGL, and D

ISSN:0148-7280    

DOI:10.1002/mrd.1120160409

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY

ISSN:0148-7280    

DOI:10.1002/mrd.1120160410

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY

ISSN:0148-7280    

DOI:10.1002/mrd.1120160401

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1987

数据来源: WILEY