Of Nyd-sp15 Biological Function And Fen On The Quality Of Adult Icr Mouse Sperm Capacitation Rate Impact

Purpose: Recently a continuous studys focused on cytidine deaminase family genes, these reports suggested that cytidine deaminases were related to body immunity, virus infection, tumorigenesis and drug resistance. NYD-SP15 is a newly identified cytidine deaminase family member, its functional study has creative significances. In the present study, we investigated the in vivo cytidine deaminase activity, subcellular location in different cell cycle, potential mechanism of nucleus-cytoplasm shuttle and gene silencing effect on cell proliferation of NYD-SP15.Methods: We constructed a prokaryotic expression plasmid containing NYD-SP15 and induced it in E.coli, cytidine deaminase activity of NYD-SP15 was analyzed by calculating the Rifampin resistant rate; subcellular location in different cell cycle was examined by constructing and infecting a NYD-SP15-GFP fusion lentivirus vector into MCF7 cell; potential nuleus export mechanism was identified by inhibiting CRM1 protein which was able to recogize and mediate nuleus export of NES-containing proteins; effect of NYD-SP15 gene scilencing was determined by constructing and infecting a NYD-SP15 RNA interfering lentivirus vector into MCF7 cell, cell cycle was detected by flow cytometry.Results: Expression in E.coli resulted in 5-fold increase of Rifampin resistant rate in ung deficiency strain BW310, but there is no DNA mutation hot-spot in rpoB gene was found. The subcellular location of NYD-SP15 in MCF7 cells revealed 3 forms: nucleus-rich, cytoplasm-rich and diffused distribution in the whole cell. The ratio of the three forms changed accord with the cell cycle, and the cytoplasm-riched cells reached its maximum ratio in metaphase while the diffused distribution reached its minimnm perportion. After LMB treatment, we found that some cell further enriched in the nuleus. Detected by flow cytometry, some of the NYD-SP15 gene scilencing cells were blocked in the S phase.Conclution: NYD-SP15 has cytidine deaminase activity in vivo; its subcellular location shuttles between nucleus and cytoplasm, and the diffused distribution pattern mainly accurs in the metaphase; CRM1-mediated nucleus exporting is a potential mechanism of the shuttle; NYD-SP15 gene scilencing cells were blocked in the S phase also suggested us that NYD-SP15 probabaly participated in the progress of DNA duplication. Purpose: Fenvalerate (FEN) has been demonstrated to be a reproductive toxicant in humans and rodents. However, little is known about whether short-term exposure to low-dose FEN produces reproductive toxicity.Methods: We administered FEN (0.009375, 0.1875, 3.750, or 45.00 mg/kg/d by gavage for 30 d) to male ICR mice and compared reproductive toxicity parameters between groups receiving different concentrations of FEN. Reproductive toxicity was evaluated by computer-assisted semen quality analysis (CASA), chlortetracycline (CTC) assay, and histopathology.Results: The sperm morphology and testis histology of FEN-exposed mice (all doses) were similar to that in control mice. Exposure to FEN at a concentration of 0.1875 mg/kg/d decreased sperm path straightness (STR) and linearity (LIN) (both Ps < 0.05), but had no significant impact on average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), lateral amplitude (ALH), beat cross frequency (BCF), or progressive motility (MOT). FEN reduced the rate of mouse sperm capacitation in a dose-dependent manner.Conclution: The present results demonstrate that exposure to low-dose FEN for 30 d reduces semen quality and sperm capacitation in adult mice.