The partial head decondensation test is a new, quick method to assess acrosome status in human spermatozoa.
OBJECTIVE
To develop a fast method for assessing acrosome status in human spermatozoa.


METHODS
Development of a new in vitro test to assess acrosome reaction in human spermatozoa.


METHODS
Academic medical institution.


METHODS
Normozoospermic subjects.


METHODS
Spermatozoa were isolated from fresh semen samples, capacitated, and stimulated or not with P or ionomycin. Acrosome reactions were evaluated by phase-contrast microscopy after a brief sperm incubation in a decondensing solution. The results were compared with those obtained by scanning electron microscopy and fluoresceinated lectin staining.


METHODS
Percentage of intact acrosomes.


RESULTS
The new procedure allowed intact acrosomes to be easily identified and quantified by phase-contrast microscopy. In unstimulated and ionomycin-treated spermatozoa, a very good agreement was found among the new test, scanning electron microscopy, and fluoresceinated lectin staining. In P-treated spermatozoa, the proposed method allowed a significantly higher percentage of reacted acrosomes to be resolved, likely due to its ability to detect the very initial stages of the acrosome reaction.


CONCLUSIONS
The new test allows acrosome-intact and acrosome-reacted spermatozoa to be unambiguously singled out and quantified. The method is rapid, reliable, sensitive, and easy to perform, which makes it of profitable use in both basic research and diagnostic practice.
Silvestroni
Leopoldo
L
University of Rome La Sapienza, and National Institute of Health, Rome, Italy. leopoldo.silvestroni@uniroma1.it
Mantovani
Alberto
A
Palleschi
Simonetta
S
D020101:Acrosome Reaction; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D008855:Microscopy, Electron, Scanning; D008858:Microscopy, Phase-Contrast; D037121:Plant Lectins; D011374:Progesterone / Q000031:analogs & derivatives / Q000494:pharmacology; D012016:Reference Values; D012680:Sensitivity and Specificity; D013077:Sperm Head / Q000502:physiology* / Q000648:ultrastructure*; D013094:Spermatozoa / Q000187:drug effects
C485330:4-pregnene-3,20-dione; D005456:Fluorescent Dyes; D007476:Ionophores; D037121:Plant Lectins; C036292:pea lectin; D011374:Progesterone; D015759:Ionomycin; D016650:Fluorescein-5-isothiocyanate