Use of diamide-acridine orange fluorescence staining to detect aberrant protamination of human-ejaculated sperm nuclei.
OBJECTIVE
To investigate the influence of human sperm nuclear chromatin on fertilization.


METHODS
Prospective study.


METHODS
Assisted reproductive technology unit at a university teaching hospital.


METHODS
Fifty men starting an IVF-ET program.


METHODS
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.


METHODS
Usefulness of diamide-acridine orange in analysis of human sperm nuclear chromatin and fertilization ability.


RESULTS
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.


CONCLUSIONS
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.
Katayose
Haruo
H
Department of Obstetrics and Gynecology, School of Medicine, Fukushima Medical University, Fukushima, Japan. katayose@fmu.ac.jp
Yanagida
Kaoru
K
Hashimoto
Shinako
S
Yamada
Hidekazu
H
Sato
Akira
A
D000165:Acridine Orange / Q000737:chemistry*; D002843:Chromatin / Q000737:chemistry; D003958:Diamide / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004624:Embryo Transfer; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D007248:Infertility, Male / Q000473:pathology*; D008297:Male; D008856:Microscopy, Fluorescence; D009698:Nucleoproteins / Q000378:metabolism; D011247:Pregnancy; D011446:Prospective Studies; D011479:Protamines / Q000737:chemistry; D012044:Regression Analysis; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000002:abnormalities / Q000166:cytology* / Q000502:physiology; D013194:Staining and Labeling / Q000379:methods
D002843:Chromatin; D005456:Fluorescent Dyes; D009698:Nucleoproteins; D011479:Protamines; D003958:Diamide; D000165:Acridine Orange