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We investigated whether the small litter size in the 129 inbred mouse strain results from a reduction in oocyte fertilizability. Sensitivity of the zona pellucida to α-chymotrypsin was examined for oocytes collected at 14 h (shortly after ovulation), 17 h, and 20 h after hCG injection. Passage of spermatozoa through the zona pellucida (using an in vitro fertilization (IVF) technique) and the density of cortical granules were examined for oocytes collected at 14 and 17 h after hCG injection. The capability of the oolemma to fuse with the sperm plasma membrane was also evaluated by IVF using zona-free eggs. The zona pellucida became markedly resistant to the enzyme 17 h after hCG injection. IVF rates significantly decreased at this time. In addition, there was a significant reduction in the density of cortical granules. When zona-free oocytes were inseminated, high fertilization rates were obtained at both 17 and 14 h after hCG injection. These results indicate that accelerated modification of the zona pellucida primarily causes a decreased fertilizability of oocytes in 129 mice, resulting in the low reproductive performance of this strain.
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http://dx.doi.org/10.1017/S0967199410000481 | DOI Listing |
Front Biosci (Landmark Ed)
August 2025
Department of Integrated Applied Life Science, Integrated Graduate School of Medicine, Engineering, and Agricultural Sciences, University of Yamanashi, 400-8510 Yamanashi, Japan.
Background: Lysosomes serve not only in the degradation of cellular components but also as calcium (Ca) stores. In this study, we investigated the effects of trans-Ned19, an inhibitor of lysosomal calcium channels known to block two-pore channels (TPCs), on fertilization and oocyte activation in mice.
Methods: Pronuclear formation was assessed via Hoechst 33342 staining, cortical granule release was evaluated using agglutinin-fluorescein isothiocyanate (LCA-FITC) staining, intracellular Ca levels were monitored with Cal-520 AM, and sperm motility was analyzed using a sperm motility analysis system (SMAS).
Biol Reprod
September 2025
Département des sciences animales, Faculté des sciences de l'agriculture et de l'alimentation, Université Laval, Québec, Qc, Canada.
Deep 3D imaging of oocytes shows several difficulties. Their large size, spherical shape causes depth-dependent artefactual shadow in the middle, resulting from refractive index mismatches induced by turbid organelles and lipid droplets. These mismatches lead to optical aberrations, increasing the laser spot size at the confocal pinhole plan and causing significant attenuation of fluorescence intensity making difficult to clearly image fine structures such as the transzonal projections (TZPs) connecting cumulus cells and the oocyte.
View Article and Find Full Text PDFMutat Res Rev Mutat Res
September 2025
Department of Molecular Biology and Biochemistry, Guru Nanak Dev University, Amritsar, Punjab 143005, India. Electronic address:
The success of Assisted Reproductive Technologies (ART), such as IVF and ICSI, relies heavily on the health of the oocyte, with abnormalities in oocyte morphology often leading to ART failure. The zona pellucida (ZP), an extracellular matrix surrounding the oocyte, plays a crucial role in sperm-egg recognition, species-specific fertilization, and protecting the embryo until implantation. This article investigates the impact of single nucleotide polymorphisms (SNPs) in the genes encoding ZP glycoproteins (hZP1, hZP2, hZP3, and hZP4) on fertility.
View Article and Find Full Text PDFReprod Med Biol
September 2025
University Farm Faculty of Agriculture, Utsunomiya University Tochigi Japan.
Purpose: In piezo-ICSI, the first polar body (PB) of a metaphase II (MII) oocyte is generally oriented in the 6 or 12 o'clock position relative to sperm injection at 3 o'clock. However, the ooplasmic cell membrane may be damaged during drilling of the zona pellucida by piezo pulses. Here, we tested a new piezo-ICSI method in which the PB is set at the 2 or 4 o'clock position, so that zona drilling is performed through the widest position in the perivitelline space (para-PB piezo).
View Article and Find Full Text PDFMethods Protoc
August 2025
Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, 165 00 Prague, Czech Republic.
The initial molecular events mediating mammalian sperm binding to the zona pellucida (ZP) of the oocyte are highly complex and still not fully elucidated. Recent advances have identified multiple candidate sperm surface proteins, often functioning as part of high-molecular-weight complexes that mediate this critical fertilization event in a species-specific and coordinated manner. To address a significant gap in the literature, we provide an in-depth overview of the functional assays employed to investigate sperm-ZP interactions, emphasizing their underlying principles, potential applications, and key methodological strengths and limitations.
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