OBJECTIVE: The oligosaccharide moieties of glycoproteins and proteoglycans have a vital function in blastocyst differentiation. Concanavalin (ConA), a lectin, is known to bind on the preimplantation embryos, especially on blastocyst. In this study, we investigated whether ConA can modulate the trophoblast development and about the regulating mediator. Also, we investigated whether expansion is enough for hatching procession of the mouse blastocyst. METHOD: Embryos were collected at 72 h post hCG injection and chemicals were treated after 24 h (96 hr post hCG injection). ConA or calcium ionophore A23187 were exposed to blastocyst and than analysis the developmental process for 48 hr. Intracellular free-Ca2+ concentration in trophectoderm was measured with confocal laser microscope after exposing to ConA or calcium ionophore A23187. ConA-pretreated blastocyst exposed to the calcium ionophore A23187 and then analyzed the developmental process. Otherwise ouabain was treated to the blastocyst to block the Na+/K+-ATPase activity. RESULTS: In contrast to the control blastocyst, the ConA-exposed blastocysts developed beyond the expansion stage with significantly high rate (90.4%) at 12 h post administration. ConA induced an increase the intracellular Ca2+ concentration in trophectoderm. Calcium ionophore A23187 also stimulated expansion of blastocyst. Most of the control blastocysts developed to the hatching stage at 144 h post hCG injection. However, strongly 65% of the ConA-exposed embryos were arrested at expanded stage at same time point. The developmental progression rates to hatching stage of both ConA- and calcium ionophore A23187-expose blastocysts were significantly lower than that of the control. However ConA-pretreated embryos developed to the hatching stage like control embryos. Ouabain showed a tendency to delayed the progress to expansion stage but did not inhibit the development to the hatching stage. CONCLUSION: ConA-mediated expansion is the result of the increase of intracellular free-calcium in blastocyst stage embryo. It is suspected that expansion of the blasocyst is a essential indirect factor in hatching and the calcium may triggering the cellular mechanisms for the both expansion and hatching progression.
Animals ; Blastocyst* ; Calcimycin ; Calcium* ; Concanavalin A* ; Embryonic Structures ; Glycoproteins ; Mice* ; Ouabain ; Proteoglycans ; Trophoblasts

Uterine cells carry out proliferation and differentiation for preparation the embryonic implantation during pregnancy. Therefore regulation of the cell proliferation is an essential step for uterine preparation, but there is not much information about the proliferation related genes in pregnant uterus. To identify these implantation specific genes, a PCR-select cDNA subtraction method was employed and got a few genes. One of the identified genes is a novel gene encoding oral tumor suppressor doc-1. To detect the doc-1 expression on the pregnant uterus, dot blotting, RT-PCR, and in situ hybridization were employed. Dot blotting revealed that doc-1 mRNA expression increase after implantation. During normal pregnancy, doc-1 mRNA expression was detected as early as day 1 of pregnancy with RT-PCR. Its expression was increased about 15 times after embryonic implantation. doc-1 transcript was localized in luminal epithelial cells but it was very faint during preimplantation. After starting the implantation, it localized in the stromal cells; heightened expression of doc-1 correlates with intense stromal cell proliferation surrounding the implanting blastocyst on day 6 morning. However in the decidualized cells, the intensity of localized doc-1 mRNA was weak. From those results, it is revealed that doc-1 express at pregnant uterus of the mouse. In addition it is suggested that doc-1 is the gene regulating the proliferation of the luminal epithelial cells and stromal cells during early implantation and decidualization.
Animals ; Blastocyst ; Cell Proliferation ; DNA, Complementary ; Epithelial Cells ; Genes, vif ; In Situ Hybridization ; Mice* ; Phenobarbital ; Pregnancy ; RNA, Messenger ; Stromal Cells ; Uterus*

Uterine cells carry out proliferation and differentiation for preparation the embryonic implantation during pregnancy. Therefore regulation of the cell proliferation is an essential step for uterine preparation, but there is not much information about the proliferation related genes in pregnant uterus. To identify these implantation specific genes, a PCR-select cDNA subtraction method was employed and got a few genes. One of the identified genes is a novel gene encoding oral tumor suppressor doc-1. To detect the doc-1 expression on the pregnant uterus, dot blotting, RT-PCR, and in situ hybridization were employed. Dot blotting revealed that doc-1 mRNA expression increase after implantation. During normal pregnancy, doc-1 mRNA expression was detected as early as day 1 of pregnancy with RT-PCR. Its expression was increased about 15 times after embryonic implantation. doc-1 transcript was localized in luminal epithelial cells but it was very faint during preimplantation. After starting the implantation, it localized in the stromal cells; heightened expression of doc-1 correlates with intense stromal cell proliferation surrounding the implanting blastocyst on day 6 morning. However in the decidualized cells, the intensity of localized doc-1 mRNA was weak. From those results, it is revealed that doc-1 express at pregnant uterus of the mouse. In addition it is suggested that doc-1 is the gene regulating the proliferation of the luminal epithelial cells and stromal cells during early implantation and decidualization.
Animals ; Blastocyst ; Cell Proliferation ; DNA, Complementary ; Epithelial Cells ; Genes, vif ; In Situ Hybridization ; Mice* ; Phenobarbital ; Pregnancy ; RNA, Messenger ; Stromal Cells ; Uterus*

The present study was designed to define the role of prostaglandin in the development and hatching of mouse embryo. The effects of indomethacin, an inhibitor of prostaglandin synthesis, on the development and hatching of morula and blastocyst were examined. In early morula stage, embryos were degenerated significantly at 100 muM and 200 muM indomethacin. However, :he viability of embryos was not influenced by concentration in any other embryonic stages. In all embryonic stages, the hatching was suppressed with concentration dependent manner, but expansion was not suppressed. Particularly, in 84h embryos post hCG injection, the hatching was suppressed significantly compared with post hCG 72h or 96h embryos. When embryos were treated with 100 muM indomethacin for a specific time (12h) in according to the development stage, the hatching was suppressed all groups. These suppressional effect was decreased as embryonic development stage was progressed. However, the expansion was not affected in all treatment group. This study suggests that hatching-related metabolic substances are synthesized from morula stage and intraembryonic signaling mediated prostaglandin was important for development and hatching of mouse embryo.
Animals ; Blastocyst ; Embryonic Development ; Embryonic Structures* ; Female ; Indomethacin* ; Mice* ; Morula ; Pregnancy

It has been revealed that multiple cohorts of tertiary follicles develop during some animal estrous cycle and the human menstrual cycle. To reach developmental competence, oocytes need the support of somatic cells. During embryogenesis, the primordial germ cells appear, travel to the gonadal rudiments, and form follicles. The female germ cells develop within the somatic cells of the ovary, granulosa cells, and theca cells. How the oocyte and follicle cells support each other has been seriously studied. The latest technologies in genes and proteins and genetic engineering have allowed us to collect a great deal of information about folliculogenesis. For example, a few web pages (http://www.ncbi.nlm.nih.gov; http://mrg.genetics.washington.edu) provide access to databases of genomes, sequences of transcriptomes, and various tools for analyzing and discovering genes important in ovarian development. Formation of the antrum (tertiary follicle) is the final phase of folliculogenesis and the transition from intraovarian to extraovian regulation. This final step coordinates with the hypothalamic-pituitary-ovarian axis. On the other hand, currently, follicle physiology is under intense investigation, as little is known about how to overcome women's ovarian problems or how to develop competent oocytes from in vitro follicle culture or transplantation. In this review, some of the known roles of hormones and some of the genes involved in tertiary follicle growth and the general characteristics of tertiary follicles are summarized. In addition, in vitro culture of tertiary follicles is also discussed as a study model and an assisted reproductive technology model.
Animals ; Axis, Cervical Vertebra ; Cohort Studies ; Embryonic Development ; Estrous Cycle ; Female ; Genetic Engineering ; Genome ; Germ Cells ; Gonads ; Granulosa Cells ; Hand ; Humans ; Menstrual Cycle ; Mental Competency ; Oocytes ; Ovary ; Pregnancy ; Proteins ; Reproductive Techniques, Assisted ; Theca Cells ; Transcriptome ; Transplants

No abstract available.
Aortic Aneurysm* ; Humans ; Rupture*

OBJECTIVES: To examine the in vitro interactions of blastocyst attachment using type I collagen. MATERIALS AND METHODS: ICR mice were used and follicular growth was stimulated by pregnant mare serum gonadotropin and human chorionic gonadotropin. On day 4 of pregnancy, the uteri were removed and blastocysts were flushed. Mixtures of 1mL sterile water, 0.5mL DMEM, 2mL type collagen solution and 0.5mL 0.1M NaOH were prepared and transferred to an incubator where the collagen solution polymerized. Blastocysts were transferred to dishes previously coated with type I collagen. CMRL 1066 was used as the basic culture medium. It was supplemented with 1mM glutamine and 1mM sodium pyruvate plus 50 IU/ml penicillin and 50 mg/ml streptomycin. During the first 4 days the culture medium was supplemented with 20% fetal calf serum and thereafter with 20% heat inactivated human cord serum. All blastocysts were initially cultured for 2 days without media change. After 2 days, fresh medium was renewed daily. The stages of embryo growth were examined and recorded everyday under a dissecting microscope and classified according to the standard in vivo criteria set forth by Witschi. RESULTS: By 48h, nearly all blastocysts had attached to the surface of collagen pad. Following adhesion to the collagen pad, the blastocysts maintained their 3-dimensional integrity in contrast to control. The embryos in collagen pad were not flattening and kept polarity and spherical shape during culture. The polar trophoblast invaded the type I collagen downward unlike the horizontal growth in control. In the developmental stage of mouse blastocyst, there were significant differences between control and type I collagen group during day 4 and 5 culture. CONCLUSION: Blastocyst development was better in type I collagen group than control. Therefore, in vitro culture study using type I collagen could provide improved model for the establishment of blastocyst implantation study.
Animals ; Blastocyst ; Chorionic Gonadotropin ; Collagen ; Collagen Type I* ; Embryo Implantation ; Embryonic Structures* ; Female ; Glutamine ; Gonadotropins ; Hot Temperature ; Humans ; Incubators ; Mice* ; Mice, Inbred ICR ; Penicillins ; Polymers ; Pregnancy ; Pyruvic Acid ; Sodium ; Streptomycin ; Trophoblasts ; Uterus ; Water

No abstract available.
Humans ; Pregnancy, Twin*

No abstract available.
Abscess* ; Drainage*

No abstract available.