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Acridine Orange

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Haruo Katayose – One of the best experts on this subject based on the ideXlab platform.

  • Use of diamide-Acridine Orange fluorescence staining to detect aberrant protamination of human-ejaculated sperm nuclei.
    Fertility and Sterility, 2003
    Co-Authors: Haruo Katayose, Kaoru Yanagida, Shinako Hashimoto, Hidekazu Yamada, Akira Sato

    Abstract:

    Abstract Objective To investigate the influence of human sperm nuclear chromatin on fertilization. Design Prospective study. Setting Assisted reproductive technology unit at a university teaching hospital. Patient(s) Fifty men starting an IVF-ET program. Intervention(s) Epifluorescent microscopic observation of human-ejaculated sperm nuclei stained with diamide–Acridine Orange. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) analysis of extracted sperm nucleoproteins. Main outcome measure(s) Usefulness of diamide–Acridine Orange in analysis of human sperm nuclear chromatin and fertilization ability. Result(s) There was no correlation between the semen parameters and the diamide–Acridine Orange observation. A positive correlation was observed between the fertilization rate after conventional IVF and the green-type increase ratio (percentage of green-pattern sperm after diamide–Acridine Orange staining/percentage of green-pattern sperm after Acridine Orange staining). Furthermore, it was suggested by SDS-PAGE that structural differences were noticed between the fertile men and the men with sperm immaturity diagnosed after diamide–Acridine Orange staining. Conclusion(s) Diamide–Acridine Orange staining was a more precise method for detecting chromatin abnormalities in human-ejaculated sperm and evaluating fertilization ability than Acridine Orange staining alone. This method can be used as a diagnostic tool to assess the fertilization ability of human-ejaculated spermatozoa before IVF procedures.

  • Thiol-disulfide status and Acridine Orange fluorescence of mammalian sperm nuclei.
    Journal of andrology, 1992
    Co-Authors: Nechama S. Kosower, Haruo Katayose, Ryuzo Yanagimachi

    Abstract:

    The relationship between thiol-disulfide status and Acridine Orange fluorescence of testicular, epididymal, and ejaculated spermatozoa in several mammalian species was investigated. Spermatozoa were fixed with acetic alcohol, stained with Acridine Orange, and examined with a fluorescence microscope. The majority of the nuclei of testicular spermatozoa of the hamster, mouse, and rabbit exhibited red Acridine Orange fluorescence. The proportion of sperm nuclei with red Acridine Orange fluorescence decreased as the spermatozoa descended the epididymis. Red Acridine Orange fluorescence was replaced by green Acridine Orange fluorescence. The site in the epididymis where 100% of the nuclei exhibited green fluorescence was the distal caput in the mouse, the corpus in the rabbit, and the proximal cauda in the hamster. In semen samples from men with proven fertility, normal semen parameters, or both, about 60% to 90% of the nuclei exhibited green Acridine Orange fluorescence. The proportion of sperm nuclei exhibiting green Acridine Orange fluorescence was higher in the spermatozoa pellet (containing highly motile spermatozoa) obtained by centrifugation through a Percoll gradient. From experiments using disulfide-reducing, thiol-oxidizing and thiol-detecting agents, we concluded that sperm nuclei fluoresce red when they are treated with acid while their DNA-associated protamines are poor in disulfides. Under such conditions, DNA is vulnerable to denaturation. Acridine Orange binds to denatured (single-stranded) DNA as aggregates and emits red fluorescence. In contrast, when sperm nuclei are treated with acid while their DNA-associated protamines are rich in disulfides, DNA is resistant to denaturation. Acridine Orange binds to native (double-stranded) DNA as a monomer and emits green fluorescence.(ABSTRACT TRUNCATED AT 250 WORDS)

Ryuzo Yanagimachi – One of the best experts on this subject based on the ideXlab platform.

  • Thiol-disulfide status and Acridine Orange fluorescence of mammalian sperm nuclei.
    Journal of andrology, 1992
    Co-Authors: Nechama S. Kosower, Haruo Katayose, Ryuzo Yanagimachi

    Abstract:

    The relationship between thiol-disulfide status and Acridine Orange fluorescence of testicular, epididymal, and ejaculated spermatozoa in several mammalian species was investigated. Spermatozoa were fixed with acetic alcohol, stained with Acridine Orange, and examined with a fluorescence microscope. The majority of the nuclei of testicular spermatozoa of the hamster, mouse, and rabbit exhibited red Acridine Orange fluorescence. The proportion of sperm nuclei with red Acridine Orange fluorescence decreased as the spermatozoa descended the epididymis. Red Acridine Orange fluorescence was replaced by green Acridine Orange fluorescence. The site in the epididymis where 100% of the nuclei exhibited green fluorescence was the distal caput in the mouse, the corpus in the rabbit, and the proximal cauda in the hamster. In semen samples from men with proven fertility, normal semen parameters, or both, about 60% to 90% of the nuclei exhibited green Acridine Orange fluorescence. The proportion of sperm nuclei exhibiting green Acridine Orange fluorescence was higher in the spermatozoa pellet (containing highly motile spermatozoa) obtained by centrifugation through a Percoll gradient. From experiments using disulfide-reducing, thiol-oxidizing and thiol-detecting agents, we concluded that sperm nuclei fluoresce red when they are treated with acid while their DNA-associated protamines are poor in disulfides. Under such conditions, DNA is vulnerable to denaturation. Acridine Orange binds to denatured (single-stranded) DNA as aggregates and emits red fluorescence. In contrast, when sperm nuclei are treated with acid while their DNA-associated protamines are rich in disulfides, DNA is resistant to denaturation. Acridine Orange binds to native (double-stranded) DNA as a monomer and emits green fluorescence.(ABSTRACT TRUNCATED AT 250 WORDS)

Akira Sato – One of the best experts on this subject based on the ideXlab platform.

  • Use of diamide-Acridine Orange fluorescence staining to detect aberrant protamination of human-ejaculated sperm nuclei.
    Fertility and Sterility, 2003
    Co-Authors: Haruo Katayose, Kaoru Yanagida, Shinako Hashimoto, Hidekazu Yamada, Akira Sato

    Abstract:

    Abstract Objective To investigate the influence of human sperm nuclear chromatin on fertilization. Design Prospective study. Setting Assisted reproductive technology unit at a university teaching hospital. Patient(s) Fifty men starting an IVF-ET program. Intervention(s) Epifluorescent microscopic observation of human-ejaculated sperm nuclei stained with diamide–Acridine Orange. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) analysis of extracted sperm nucleoproteins. Main outcome measure(s) Usefulness of diamide–Acridine Orange in analysis of human sperm nuclear chromatin and fertilization ability. Result(s) There was no correlation between the semen parameters and the diamide–Acridine Orange observation. A positive correlation was observed between the fertilization rate after conventional IVF and the green-type increase ratio (percentage of green-pattern sperm after diamide–Acridine Orange staining/percentage of green-pattern sperm after Acridine Orange staining). Furthermore, it was suggested by SDS-PAGE that structural differences were noticed between the fertile men and the men with sperm immaturity diagnosed after diamide–Acridine Orange staining. Conclusion(s) Diamide–Acridine Orange staining was a more precise method for detecting chromatin abnormalities in human-ejaculated sperm and evaluating fertilization ability than Acridine Orange staining alone. This method can be used as a diagnostic tool to assess the fertilization ability of human-ejaculated spermatozoa before IVF procedures.