use of the acridine orange test and the TUNEL assay to assess the integrity of freeze-dried bovine spermatozoa DNA
The ability to detect nuclear damage is an important tool for the development of sperm preservation methods. We used the acridine orange test (AOT) and the terminal deoxynucleotidyl transferase-mediated dUDP nick-end labeling (TUNEL) assay to assess the DNA status of sperm cells preserved with diffe...
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Published in | Genetics and molecular research Vol. 6; no. 1; pp. 94 - 104 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Brazil
01.01.2007
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Abstract | The ability to detect nuclear damage is an important tool for the development of sperm preservation methods. We used the acridine orange test (AOT) and the terminal deoxynucleotidyl transferase-mediated dUDP nick-end labeling (TUNEL) assay to assess the DNA status of sperm cells preserved with different lyophilization media. The AOT did not detect any differences between different lyophilization media. However, differences in DNA integrity were observed among treatments with the TUNEL assay, suggesting that TUNEL is a more sensitive method to evaluate sperm DNA. The use of TCM 199 and 10% FCS as a lyophilization medium resulted in 14% of the cells with DNA fragmentation in TUNEL test. The AOT indicated only 4% of the cells with chromatin damage, with this same treatment, with no significant differences when compared to the other treatments. The degree of DNA fragmentation was negatively related to fertilizing potential, as sperm DNA damage was inversely correlated with pro-nucleus formation. The TUNEL assay was found to be an efficient method to detect DNA damage in sperm, and it could be used as a tool to predict male fertility. |
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AbstractList | The ability to detect nuclear damage is an important tool for the development of sperm preservation methods. We used the acridine orange test (AOT) and the terminal deoxynucleotidyl transferase-mediated dUDP nick-end labeling (TUNEL) assay to assess the DNA status of sperm cells preserved with different lyophilization media. The AOT did not detect any differences between different lyophilization media. However, differences in DNA integrity were observed among treatments with the TUNEL assay, suggesting that TUNEL is a more sensitive method to evaluate sperm DNA. The use of TCM 199 and 10% FCS as a lyophilization medium resulted in 14% of the cells with DNA fragmentation in TUNEL test. The AOT indicated only 4% of the cells with chromatin damage, with this same treatment, with no significant differences when compared to the other treatments. The degree of DNA fragmentation was negatively related to fertilizing potential, as sperm DNA damage was inversely correlated with pro-nucleus formation. The TUNEL assay was found to be an efficient method to detect DNA damage in sperm, and it could be used as a tool to predict male fertility. |
Author | Martins, C.F Dode, M.N Rumpf, R Báo, S.N |
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SubjectTerms | acridine orange animal breeding Animals Cattle Chromatin - chemistry Chromatin - metabolism Coloring Agents DNA - chemistry DNA - metabolism DNA damage DNA Fragmentation fertilization (reproduction) Flow Cytometry Freeze Drying In Situ Nick-End Labeling Male Nellore Nucleic Acid Conformation Protein Conformation Semen Preservation - adverse effects Semen Preservation - methods spermatozoa Spermatozoa - chemistry Staining and Labeling terminal deoxynucleotidyl transferase-mediated dUDP nick-end labeling assay |
Title | use of the acridine orange test and the TUNEL assay to assess the integrity of freeze-dried bovine spermatozoa DNA |
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