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*

The Spindle-view, a specialized instrument for observing spindle image, was applied to observe the meiotic spindles of vitro matured porcine oocytes at 36, 42, 44, 48h, and enucleation from porcine, comparing to the previously methods (McGrath-Solter's method and two-step-squeezing method) in the enucleated. The results showed that: (1) there was no noticeable differences at vicinity of spindle images and 1st polar body among in vitro matured porcine oocytes at 40-48 h under the instrument; (2) Spindle-view is suitable for the observation of meiotic spindles of matured oocytes and enucleation from porcine; the modified Spindle-view method for enucleation is significantly better than McGrath-Solter' s method and two-step-squeezing method in the enucleated rates (95.5%, 42.1%, 74.2%, P < 0.0l) of absolutely removing nuclei matter; (3) the spindle images could be used to monitor the oocyte qualities.
Animals ; Cell Nucleus ; ultrastructure ; Cells, Cultured ; Cytological Techniques ; Female ; Nuclear Transfer Techniques ; Oocytes ; cytology ; Spindle Apparatus ; ultrastructure ; Swine

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 investigate the mechanism of oocyte aneuploidy formation.

METHODSThe unfertilized oocytes were fixed 1-3 days after oocyte retrieval. Multicolor fluorescence in situ hybridization (M-FISH) was performed according to the Vysis protocol to check the chromosome status in oocytes by using centromeric enumerator probes for chromosomes 16, 18 and locus-specific probes for chromosomes 13, 21 and 22.

RESULTS47% oocytes were found to be normal, while 53 % oocytes were abnormal. Nondisjunction was found in 22(18%) oocytes, unbalanced predivision of chromatids in 15(12%) oocytes, balanced predivision of chromatids in 45(36%) oocytes. The balanced predivision rate in oocytes aged in vitro>24h was much higher than that in oocytes aged in vitro

CONCLUSIONBoth nondisjunction, balanced and unbalanced predivision of chromatids are involved in the oocyte aneuploidy formation. Balanced predivision is related to the time in culture.

Adult ; Aneuploidy ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; Oocytes ; ultrastructure

The nuclear pore complex (NPC), one of the largest protein complexes in eukaryotes, serves as a physical gate to regulate nucleocytoplasmic transport. Here, we determined the 8 Å resolution cryo-electron microscopic (cryo-EM) structure of the outer rings containing nuclear ring (NR) and cytoplasmic ring (CR) from the Xenopus laevis NPC, with local resolutions reaching 4.9 Å. With the aid of AlphaFold2, we managed to build a pseudoatomic model of the outer rings, including the Y complexes and flanking components. In this most comprehensive and accurate model of outer rings to date, the almost complete Y complex structure exhibits much tighter interaction in the hub region. In addition to two copies of Y complexes, each asymmetric subunit in CR contains five copies of Nup358, two copies of the Nup214 complex, two copies of Nup205 and one copy of newly identified Nup93, while that in NR contains one copy of Nup205, one copy of ELYS and one copy of Nup93. These in-depth structural features represent a great advance in understanding the assembly of NPCs.
Animals ; Artificial Intelligence ; Cryoelectron Microscopy ; Nuclear Pore/ultrastructure* ; Oocytes/metabolism* ; Xenopus laevis

We report the successful outcome of intracytoplasmic sperm injection (ICSI) treatment in two siblings with familial globozoospermia. After controlled ovarian hyperstimulation and oocyte pick-up, retrieved oocytes were mechanically activated before ICSI and a fertilization rate of 33.3% was achieved in the first case. The second couple underwent ICSI without oocyte activation and a 9.1% fertilization rate was obtained. The transfer of two grade I embryos in the first couple and one grade I embryo in the second couple resulted in clinical pregnancies with healthy livebirths. It was concluded that the main problem of cases with globozoospermia is a low fertilization rate, and even though ICSI and oocyte activation can increase this rate it is not necessarily needed to achieve a pregnancy.
Acrosome ; Adult ; Female ; Humans ; Infertility, Male ; physiopathology ; therapy ; Live Birth ; Male ; Oocyte Retrieval ; Oocytes ; physiology ; Pregnancy ; Sperm Head ; ultrastructure ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; abnormalities ; ultrastructure ; Treatment Outcome

OBJECTIVETo evaluate the morphology and proliferation of follicles from cryopreserved human ovarian tissue by vitrification.

METHODSOvarian biopsy specimens were taken from 12 patients. The specimens were randomly distributed into fresh group (Group A) and vitrification group (Group B). Histological examination and ultrastructural observation were performed after cryopreservation. Both were embedded in paraffin block and proliferating cell nuclear antigen (PCNA) was detected by immunohistochemical staining.

RESULTSThe proportions of primordial and primary follicles from Group A and Group B were 86.4%, 13.6% and 84.5%, 15.5%, respectively (P>0.05). There was no significant difference in proportions of morphologically normal primordial follicles between Group A and Group B (P>0.05); but the proportion of morphologically abnormal primary follicles was significantly higher in Group B than that in Group A (P<0.05). The ultrastructural studies showed that in histologically normal primordial follicles, there was no difference between Group A and Group B, while there were a few abnormalities of primary follicles in Group B. Granulosa cells and oocytes of primordial and primary follicles and stromal cells were positive for PCNA staining both in fresh and cryopreserved ovarian tissues; there were no differences between two groups.

CONCLUSIONVitrification is a favorable method in human ovarian cryopreservation.

Adult ; Cell Proliferation ; Cryopreservation ; methods ; Female ; Humans ; In Vitro Techniques ; Oocytes ; cytology ; Ovarian Follicle ; cytology ; ultrastructure ; Ovary ; anatomy & histology ; Vitrification

This study was conducted to evaluate the microtubule distribution following control of nuclear remodeling by treatment of bovine somatic cell nuclear transfer (SCNT) embryos with caffeine or roscovitine. Bovine somatic cells were fused to enucleated oocytes treated with either 5 mM caffeine or 150 micrometer roscovitine to control the type of nuclear remodeling. The proportion of embryos that underwent premature chromosome condensation (PCC) was increased by caffeine treatment but was reduced by roscovitine treatment (p < 0.05). The microtubule organization was examined by immunostaining beta- and gamma-tubulins at 15 min, 3 h, and 20 h of fusion using laser scanning confocal microscopy. The gamma-tubulin foci inherited from the donor centrosome were observed in most of the SCNT embryos at 15 min of fusion (91.3%) and most of them did not disappear until 3 h after fusion, regardless of treatment (82.9-87.2%). A significantly high proportion of embryos showing an abnormal chromosome or microtubule distribution was observed in the roscovitine-treated group (40.0%, p < 0.05) compared to the caffeine-treated group (22.1%). In conclusion, PCC is a favorable condition for the normal organization of microtubules, and inhibition of PCC can cause abnormal mitotic division of bovine SCNT embryos by causing microtubule dysfunction.
Animals ; Caffeine/pharmacology ; Cattle/embryology/*physiology ; Cell Nucleus/drug effects/*physiology/ultrastructure ; Female ; Fertilization in Vitro/veterinary ; Male ; Microscopy, Confocal/veterinary ; Microtubules/drug effects/*physiology/ultrastructure ; Nuclear Transfer Techniques/veterinary ; Oocytes/*physiology ; Pregnancy ; Purines/pharmacology