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NYD-SP15 FEN biological function and quality of adult ICR mice sperm capacitation and the rate of

Author: BiHuanJing
Tutor: LiJianMin
School: Nanjing Medical University
Course: Cell Biology
Keywords: Cytidine deaminase NYD-SP15 Cell Cycle Subcellular localization RNA interference Sperm Capacitation rate Endocrine disruptors Fenvalerate Computer-aided sperm analysis
CLC: Q954.43
Type: Master's thesis
Year: 2011
Downloads: 23
Quote: 0
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Abstract


Background Objective: In recent years about the cytidine deaminase gene family flocked to the research report, many of the family members in tumorigenesis, tumor resistance, immune and anti-viral infection and other biological activities exercised an important role. NYD-SP15 is what we found with cytidine deaminase domain of new genes, their function studies groundbreaking significance. This study of NYD-SP15 vivo cytidine deaminase activity in different cell cycle subcellular localization of nuclear and cytoplasmic shuttling of the possible mechanisms of gene silencing effect on cell proliferation and other studies, preliminary clarify NYD-SP15 gene function. Methods: By constructing NYD-SP15 prokaryotic expression plasmid and induce NYD-SP15 protein expressed in E. coli, according to the determination of rifampicin mutation frequency analysis of its in vivo cytidine deaminase; NYD-SP15 by building green fluorescence fusion protein lentivirus and infect MCF7 cells were analyzed in different cell cycle position; through inhibition can be specifically identified NES sequence and mediates the nuclear transport protein CRM1 protein research NYD-SP15 possible nuclear export mechanism; by building NYD-SP15 RNA interference lentiviral vectors to infect MCF7 cells, and cell cycle by flow cytometry to study the NYD-SP15 gene silencing on cell cycle. Results: NYD-SP15 induced expression in Escherichia coli ung gene inactivation can produce rifampicin resistant strain BW310 5-fold increase in the frequency, but the induction of DNA mutations in the rpoB gene in E. coli show no interest zone. NYD-SP15 in MCF7 cells, showing the positioning of three forms: the enrichment of nuclear and cytoplasmic enrichment and uniformly distributed, and the three forms of the proportion of cells with the change of cell cycle, and the M phase enriched in the cytoplasm cell ratio reaches a maximum, while uniform distribution of cells reaches a minimum. LMB treatment seen after NYD-SP15 in some cells further enriched in the nucleus. The NYD-SP15 in MCF7 cells after RNA interference, by flow cytometry revealed interference group cell cycle arrest at S phase. Conclusion: NYD-SP15 in vivo with cytidine deaminase activity; localization in cells with nuclear - cytoplasmic shuttling used, and in the M phase mainly localized in the cytoplasm; while NYD-SP15 This may be associated with the nuclear transport protein CRM1 relevant; RNA interference found cells were arrested in S phase also prompt us to NYD-SP15 may be involved in DNA replication. Objective: To observe the short-term low-dose fenvalerate (FEN) exposure on adult male ICR mice reproductive system and sperm quality, vitality, was able to rate. Methods: 50 adult male ICR mice were randomly divided into five groups, respectively 0,0.009375,0.1875,3.75 and 45mg/kg doses of continuous gavage FEN sacrificed after 30 days, mice were measured by body weight and testes weight ; using computer-aided sperm analysis (CASA) was measured before and after sperm capacitation; chlortetracycline (CTC) staining was measured rate of energy; testicular sections for HE staining of histological changes. Results: FEN treated group compared with control group, 0.1875mg/kg group after sperm capacitation motion path straightness (STR) and linearity (LIN) has decreased significantly (P lt; 0.05); 0.009375,0.1875,3.750 and 45.00 mg / kg dose group compared with the control group, sperm capacitation rate showed a highly significant dose - effect of reduced (P lt; 0.01). Conclusion: Low-dose FEN adult mice exposed to 30 days can sperm quality and sperm capacitation rate.

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CLC: > Biological Sciences > Zoology > Animal morphology > Animal Embryology ( of animals,animal viviparous learn ) > Germ cells ( gametes )
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