The ultrastructural changes of mitochondria in the ovarian oocytes from Graafian follicles, the ovulated tubal ova, and the various stages of preimplantation rabbit embryos have been observed with an electron microscope. From the ovarian oocytes to the 4-cell stage, mitochondria showed oval and round forms with a few cristae arranged concentrically and peripherally at the inner membrane. In 8-cell and 16-cell stages, mitochondria tended to change their forms to be elongated, and their sizes, and the outer membrane of the mitochondria had a tendency to become rough and irregular although there were few changes in the inner structure. In morula, some mitochondria began to show several transverse cristae proceeding into the matrix. Mitochondria rapidly increased in number at the late blastocyst stage. Matrix of mitochondria with transverse cristae found in the morula and in blastocyst stages was less dense than that of the earlier stages. The authors believe that the morphological changes of mitochondria during early embryonal development indicate the level of enzymatic activity at which this organelle is engaged in energy metabolism.
Animal ; Cell Membrane/ultrastructure ; Embryo/ultrastructure* ; Embryo Implantation ; Female ; Microscopy, Electron ; Mitochondria/ultrastructure* ; Organoids/ultrastructure ; Ovum/ultrastructure ; Rabbits

This experiment was undertaken in order to find out if there is any morphological change in oocytes and two-cell embryos whose development have been suppressed by progesterone for six hours in vitro. It can be observed that some part of the outer side of nuclear membrane of the suppressed oocytes was damaged. The number of nuclear pores has decreased in suppressed oocytes and this suggests that progesterone might suppress the transport of intermediary metabolites between cytoplasm and nucleus. Sometimes, closely packed aggregates of parallel or irregular endoplasmic reticula were observed in suppressed oocytes. Microvilli of suppresed oocytes showed signs of degradation and the perivitelline space became apparent. Thus it is presumed that the egg membrane has constricted during cultivation under progesterone in vitro. The other cell organelles such as mitochondria, multivesicular bodies, cortical granules and fibrillar lattices showed no difference in morphology between treated and control (intact) oocytes. In two-cell embryos, there was also no evident morphological change except for the fact that many vacuoles appeared clearly in suppressed embryonal cells. In brief, there was no fundamental morphological change in the oocytes and the embryonal cells exposed to progesterone for six hours even though it inhibits their development. The action of progesterone should be investigated thoroughly.
Animal ; Embryo/cytology* ; Embryo/drug effects ; Female ; In Vitro ; Mice ; Oocytes/drug effects ; Oocytes/ultrastructure* ; Ovum/ultrastructure* ; Progesterone/pharmacology*

OBJECTIVETo observe morphological characteristics and in-vitro maturation ratio of all metaphase I arrested oocytes in IVF-ET cycles.

METHODSIn two IVF-ET cycles, maturation culture in vitro was performed and we evaluated morphological characteristics of all oocytes arrested in metaphase I.

RESULTSMetaphase I arrested oocytes presented with uniform cytoplasm, compact zone and no space between oocyte and zone. Incompact association was observed between granule cell and zone without radiated structures. There was no markedly alterations in morphological characteristics after 24, 48, 72 hours of maturation culture, respectively. All oocytes were arrested in Metaphase I and no polar growed.

CONCLUSIONAll metaphase I arrested oocytes possess of unique morphological characteristics and fail to mature following current culture assay in IVF-ET cycles.

Adult ; Cells, Cultured ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Metaphase ; Oocytes ; ultrastructure

OBJECTIVETo evaluate the value of endometrial microvessel density (MVD) in assessing the endometrial receptivity during the peri-implantation period.

METHODSA total of 104 patients undergoing in vitro fertilization and embryo transfer (IVF-ET) treatment were analyzed retrospectively. The subjects were divided into clinical pregnancy group (50 cases) and nonpregnant group (54 cases) according to the IVF-ET outcome. Endometrial tissues were collected 7 days after the natural ovulation prior to IVF-ET for measurement of the endometrial MVD using electron microscopy, which was analyzed in relation to the clinical outcome of the treatment.

RESULTSThe endometrial MVD was significantly higher in the clinical pregnancy group than in the nonpregnant group [(4.12∓1.84)% vs (3.46∓1.26)%, t=-2.127, P=0.036). ROC curve analysis showed that the MVD had an area under the curve slightly over 0.5 (0.598) for predicting clinical pregnancy, suggesting a poor specificity in predicting the clinical outcome of the treatment.

CONCLUSIONIn IVF-ET cycles, the endometrial MVD during the peri-implantation period is helpful for assessing the endometrial receptivity, but the specificity remains low.

Adult ; Embryo Implantation ; physiology ; Embryo Transfer ; Endometrium ; blood supply ; physiology ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; diagnostic imaging ; physiopathology ; therapy ; Microvessels ; ultrastructure ; Retrospective Studies ; Ultrasonography

OBJECTIVETo investigate the influence of the DNA integrity of optimized sperm on the embryonic development and clinical outcomes of in vitro fertilization and embryo transfer (IVF-ET).

METHODSThis study included 605 cycles of conventional IVF-ET for pure oviductal infertility performed from January 1, 2013 to December 31, 2014. On the day of retrieval, we examined the DNA integrity of the sperm using the sperm chromatin dispersion method. According to the ROC curve and Youden index, we grouped the cycles based on the sperm DNA fragmentation index (DFI) threshold value for predicting implantation failure, early miscarriage, and fertilization failure, followed by analysis of the correlation between DFI and the outcomes of IVF-ET.

RESULTSAccording to the DFI threshold values obtained, the 605 cycles fell into four groups (DFI value < 5%, 5-10%, 10-15%, and ≥ 15%). Statistically significant differences were observed among the four groups in the rates of fertilization, cleavage, high-quality embryo, implantation, clinical pregnancy, early miscarriage, and live birth (P < 0.05), but not in the rates of multiple pregnancy, premature birth, and low birth weight (P > 0.05). DFI was found to be correlated negatively with the rates of fertilization (r = -0.32, P < 0.01), cleavage (r = -0.19, P < 0.01), high-quality embryo (r = -0.40, P < 0.01), clinical pregnancy (r = -0.20, P < 0.01), and live birth (r = -0.09 P = 0.04), positively with the rate of early miscarriage (r = 0.23, P < 0.01), but not with the rates of multiple pregnancy (r = -0.01, P = 0.83), premature birth (r = 0.04, P = 0.54), and low birth weight (r = 0.03, P = 0.62).

CONCLUSIONThe DNA integrity of optimized sperm influences fertilization, embryonic development, early miscarriage, and live birth of IVF-ET, but its correlation with premature birth and low birth weight has to be further studied.

Abortion, Spontaneous ; Chromatin ; ultrastructure ; DNA Fragmentation ; Embryo Implantation ; Embryo Transfer ; Embryonic Development ; Female ; Fertilization ; Fertilization in Vitro ; Humans ; Infertility, Female ; Male ; Pregnancy ; Pregnancy Outcome ; ROC Curve ; Spermatozoa ; cytology

Cell apoptosis induced by duck reovirus (DRV)in duck embryo fibroblasts (DEF) was ascertained by light microscope and electron microscopy, DNA Ladder, flow cytometry and fluorescent microscopy. Typical morphological apoptotic features including cell shrinkage and condensation, margination of nuclear chromatin were observed under light microscope and the formation of apoptotic bodies by electron microscopy. DNA ladder was shown by DNA fragment analysis at 24-144h post infection. Flow cytometry showed that the cell apoptosis appeared at 24h and reached it's crest-time at 72-96h, decreased at 144h. Fluorescent microscopy showed that the apoptotic cells which showed green fluorescence appeared at 24h, the number of dead cells which showed red fluorescence increased with the time went by. The results above confirmed that the apoptosis of DEF was successfully induced by DRV.
Animals ; Apoptosis ; physiology ; Cell Nucleus ; ultrastructure ; Cells, Cultured ; DNA Fragmentation ; Ducks ; Embryo, Nonmammalian ; cytology ; Fibroblasts ; cytology ; ultrastructure ; virology ; Flow Cytometry ; Host-Pathogen Interactions ; Microscopy, Electron, Transmission ; Reoviridae ; physiology

BACKGROUNDThere have been no detailed reports of the three-dimensional structure and the relationship between the external and internal vascularizations observed successively for a long duration in the rat fetus, although many authors have studied the vascular morphology of the developing brain. This study examined the three-dimensional structure of both the external and internal vascularizations of the prenatal rat telencephalon from embryonic days 12 (E12) to 20 (E20).

METHODA microvascular casting method for scanning electron microscopy (SEM) was used in this study, along with vascular staining using gold-gelatin solution-autometallography (GGS-AMG) after intravascular injection of colloidal gold, as well as hematoxylin-eosin (HE) staining for paraffin embedded specimens.

RESULTSIn GGS-AMG stains, E16 fetuses had a few short perforating cortical blood vessels (SPCVs); E17 fetuses had long perforating cortico-medullary vessels (LPCVs). Older fetuses had specific patterns of vascular networks in the cortex and the deeper subcortical part of the telencephalon. In the cortex, fine longitudinal blood vessels were connected by transverse channels. The deep telencephalon had fine blood vessels running in all directions. Using SEM, the external vascularization was already visible in E12 fetuses as arborizations of arterial branches, forming a mesh of fine vascular networks covering the telencephalon. A coralliform fine venous plexus was observed in the external vascularization of E16 fetuses. There were ring-like anastomoses and bud-like protrusions in the network of small blood vessels, most likely the angiogenesis of fetal vessels. From E12 to E16, an immature and incomplete internal vascularization began to appear. There were short blood vessels with ballooned terminals branching from the external vascularization. They penetrated the brain tissue to form networks in the superficial layer, comparable to SPCVs. In E17 to E20 fetuses, tortuous venous branches, straight arterial blood vessels, and a fine network of small blood vessels formed the external vascularization. There were fewer arterial than venous branches connecting to the fine networks of small blood vessels. LPCVs were noted at E17, at the time the white matter emerged. They branched from the external vascularization, and perpendicularly penetrated the brain surface, traversing the cortical plate, and entering into the deep brain. At E17, arterial and venous blood vessels could be clearly distinguished in the external vascularization. At E20, the cortex and white matter contained specific arrangements of networks of fine blood vessels, as seen by GGS-AMG staining.

CONCLUSIONThese findings show that the development of both the external and internal vascularization follows the development of the telencephalon. In particular, the emergence of the cortical plate and white matter on E16 and E17 influence the development of both the internal and the external vascularization. The laminal arrangement of blood vessels was not observed corresponding to the respective laminal neuronal layers.

Animals ; Blood Vessels ; embryology ; ultrastructure ; Embryo, Mammalian ; Fetus ; Microscopy, Electron, Scanning ; Rats ; Rats, Wistar ; Telencephalon ; blood supply

AIMThe production of interspecific germline chimeras between chicken and quail were attempted employing the dissociated cells derived from the blastodermal central disk (stage X) and the germinal crescent region of embryo (stage 7-8).

METHODSThe central disk (CD) of the area pellucida in chicken blastoderm (stage X) and the germinal crescent region (GCR) of embryo (stage 7-8) were dispersed and injected into the subgerminal cavity of quail blastoderm (stage X). Injected eggs were incubated for 7 days or to hatching. The donor chicken DNA was detected by the polymerase chain reaction.

RESULTSIn day-7 embryos, chicken DNA was detected in 5 gonads and 9 brains from 53 survived embryos received chicken CD cells, and 1 gonads and 6 brains from 27 survived embryos received chicken GCR. Chicken DNA was also detected from the semen of one adult male hatched from eggs received chicken GCR cells.

CONCLUSIONCD and GCR cells as the donors showed the possibility to produce the interspecific germline chimera, but further studies are needed to make necessary improvement.

Animals ; Base Sequence ; Blastoderm ; physiology ; ultrastructure ; Brain ; embryology ; Brain Chemistry ; Chick Embryo ; physiology ; Chickens ; Chimera ; DNA Primers ; DNA, Complementary ; genetics ; Embryo, Nonmammalian ; physiology ; Female ; Germ-Line Mutation ; physiology ; Male ; Ovalbumin ; genetics ; Ovary ; embryology ; Polymerase Chain Reaction ; Quail ; Testis ; embryology

OBJECTIVETo study the relationship between late mRNA and the cytopathic effect(CPE) and ultrastructural features after human cytomegalovirus (HCMV) infection in vitro.

METHODSHuman embryo fibroblast cells(HEL) were infected with HCMV AD169 strain. The expression of the HCMV late mRNA was measured by semiquantitative RT-PCR, the cytopathic effect (CPE) and the cell ultrastructure were observed by means of light microscopy and electron microscopy, respectively.

RESULTSThe HCMV late mRNA could be detected 12 hours postinfection and increased gradually, but the CPE appeared 48 hours postinfection in HEL cells. The HCMV infected cells exhibited significant mitochondrial enlargement and the number of mitochondrial ridge deletion, the cisternae lumen of endoplasmic reticulum (ER) dilation and vacuolization (at the end age). The mature nucleocapsid could be observed 96 hours postinfection.

CONCLUSIONThe ultrastructural changes have an intimate correlation with the expression of HCMV late mRNA and play an important role in the life circle of the virus. HCMV late mRNA may serve as a indicator of the clinical effect of treatment in active HCMV infection.

Cytomegalovirus ; genetics ; physiology ; Cytopathogenic Effect, Viral ; Embryo, Mammalian ; Endoplasmic Reticulum ; pathology ; virology ; Fibroblasts ; metabolism ; ultrastructure ; virology ; Humans ; Inclusion Bodies ; pathology ; virology ; Mitochondrial Swelling ; RNA, Messenger ; metabolism

OBJECTIVETo investigate the effect of cytomegalovirus (CMV) infection on actin and microfilament in human embryo fibroblast cells (HF) and its relationship with CMV replication.

METHODSCell morphology was observed after the infection of CMV. Western-blot was used to measure the expression levels of beta-actin, G-actin and F-actin proteins. CMV immediately early antigen (CMV IE) in HF cells was analyzed by indirect immunofluorescence assay. Microfilament alteration was determined by cytoskeleton fluorescence probe.

RESULTCMV IE was demonstrated in more than 95% of HF cells after infection, which was primarily located in nucleus. The shape of HF cells changed from thin shuttle like to round and thick ball like, even escaping from wall after infection by CMV. Compared with control group, the expression of G-actin protein increased at 24 h of CMV infection (0.941 +/-0.061 compared with 0.714 +/-0.119, P <0.05), then decreased at 72 h, 96 h respectively(0.218 +/-.035, 0.230 +/-0.055 compared with 0.714 +/-0.119, P <0.05). The levels of F-actin in infected cells gradually decreased at 24 h, 72 h and 96 h compared with control HF cells (0.256 +/-0.021, 0.127 +/-0.032, 0.026 +/-0.008 compared with 0.373 +/-0.050, P<0.05). In infected HF cells, microfilaments were found ruptured, arranged turbulently. Cells fused and fluorescence density of microfilament markedly reduced.

CONCLUSIONCytomegalovirus can induce alteration of actins and microfilament, which may be associated with its infection, replication and reactivity in host cells.

Actin Cytoskeleton ; metabolism ; Actins ; biosynthesis ; genetics ; Antigens, Viral ; analysis ; Cells, Cultured ; Cytomegalovirus ; Cytoskeleton ; metabolism ; Embryo, Mammalian ; Fibroblasts ; metabolism ; ultrastructure ; virology ; Humans ; Immediate-Early Proteins ; analysis