The Experts below are selected from a list of 31467 Experts worldwide ranked by ideXlab platform
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, 2003Co-Authors: Haruo Katayose, Kaoru Yanagida, Shinako Hashimoto, Hidekazu Yamada, Akira SatoAbstract: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, 1992Co-Authors: Nechama S. Kosower, Haruo Katayose, Ryuzo YanagimachiAbstract: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, 1992Co-Authors: Nechama S. Kosower, Haruo Katayose, Ryuzo YanagimachiAbstract: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, 2003Co-Authors: Haruo Katayose, Kaoru Yanagida, Shinako Hashimoto, Hidekazu Yamada, Akira SatoAbstract: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.
Raymond Julien - One of the best experts on this subject based on the ideXlab platform.
-
10N-nonyl Acridine Orange interacts with cardiolipin and allows the quantification of this phospholipid in isolated mitochondria.
European journal of biochemistry, 1992Co-Authors: Jean-michel Petit, Abderrahman Maftah, Marie-hélène Ratinaud, Raymond JulienAbstract:The Acridine Orange derivative, 10N-nonyl Acridine Orange, is an appropriate marker of the inner mitochondrial membrane in whole cells. We use membrane model systems to demonstrate that 10N-nonyl Acridine Orange binds to negatively charged phospholipids (cardiolipin, phosphatidylinositol and phosphatidylserine). The stoichiometry has been found to be 2 mol 10N-nonyl Acridine Orange/mol cardiolipin and 1 mol dye/mol phosphatidylserine or phosphatidylinositol, while, with zwitterionic phospholipids, significant binding could not be detected. The affinity constants were 2 x 10(6) M-1 for cardiolipin-10N-nonyl-Acridine-Orange association and only 7 x 10(4) M-1 for that of phosphatidylserine and phosphatidylinositol association. The high affinity of the dye for cardiolipin may be explained by two essential interactions; firstly an electrostatic interaction between the quaternary ammonium of nonyl Acridine Orange and the ionized phosphate residues of cardiolipin and secondly, hydrophobic interactions between adjacent chromophores. A linear relationship was demonstrated between the cardiolipin content of model membranes and the incorporated dye. Consequently, a convenient and rapid method for cardiolipin quantification in membranes was established and applied to the cardiolipin-containing organelle, the mitochondrion.
Luz Hernández-esquivel - One of the best experts on this subject based on the ideXlab platform.
-
Titration of cardiolipin by either 10-N-nonyl Acridine Orange or Acridine Orange sensitizes the adenine nucleotide carrier to permeability transition
Journal of Bioenergetics and Biomembranes, 2008Co-Authors: Edmundo Chávez, Noemi Garcia, Eduardo Martínez-abundis, Cecilia Zazueta, N Pavon, Luz Hernández-esquivelAbstract:Under the action of carboxyatractyloside or fatty acids, adenine nucleotide translocase switches its function from nucleotide carrier to modulator of the opening of a non-specific pore. In addition to the effect of these agents, this modification in activity is, in some way, dependent on the influence of the lipid milieu of the membrane. Cardiolipin is, among other membrane phospholipids, the one that interacts the most with the translocase. This work shows that 10- N -nonyl Acridine Orange and Acridine Orange, probes for this phospholipid, modify the sensitivity of the translocase to carboxyatractyloside, oleate, and palmitate to induce permeability transition. The results also show that these probes stimulate the release of mitochondrial cytochrome c , and increase labeling of the carrier by eosin 5-maleimide. Based on the aforementioned it is proposed that the increase in sensitivity is due to a conformational change in the translocase, induced by the binding of the probe to cardiolipin.
