Destabilization of acrosome and elastase influence mediate the release of secretory phospholipase A2 from human spermatozoa.
OBJECTIVE
To determine the cellular distribution of secretory phospholipase A(2) (sPLA(2)) in dependence on the acrosomal state and under the action of elastase released under inflammatory processes from leukocytes.


METHODS
Acrosome reaction of spermatozoa was triggered by calcimycin. Human leukocyte elastase was used to simulate inflammatory conditions. To visualize the distribution of sPLA(2) and to determine the acrosomal state, immunofluorescence techniques and lectin binding combined with confocal laser scanning fluorescence microscopy and flow cytometry were used.


RESULTS
Although sPLA(2) was detected at the acrosome and tail regions in intact spermatozoa, it disappeared from the head region after triggering the acrosome reaction. This release of sPLA(2) was associated with enhanced binding of annexin V-fluoroscein isothiocyanate (FITC) to spermatozoa surfaces, intercalation of ethidium-homodimer I, and binding of FITC-labelled concanavalin A at the acrosomal region. Spermatozoa from healthy subjects treated with elastase were characterized by release of sPLA(2), disturbance of acrosome structure, and loss of vitality.


CONCLUSIONS
The ability of spermatozoa to release secretory phospholipase A(2) is related to the acrosomal state. Premature destabilization of the acrosome and loss of sPLA(2) can occur during silent inflammations in the male genital tract. The distribution pattern of sPLA(2) in intact spermatozoa might be an additional parameter for evaluating sperm quality.
Lessig
Jacqueline
J
University of Leipzig, Medical Faculty, Institute for Medical Physics and Biophysics, Haertelstrasse 16-18, Leipzig D-04107, Germany. jacqueline.lessig@medizin.uni-leipzig.de
Reibetanz
Uta
U
Arnhold
Jurgen
J
Glander
Hans-Jurgen
HJ
D000177:Acrosome / Q000187:drug effects / Q000502:physiology*; D020101:Acrosome Reaction / Q000187:drug effects; D017304:Annexin A5 / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D004996:Ethidium; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D018613:Microscopy, Confocal; D010196:Pancreatic Elastase / Q000378:metabolism*; D010718:Phosphatidylserines / Q000378:metabolism; D054497:Phospholipases A2, Secretory / Q000378:metabolism*; D012661:Semen / Q000166:cytology / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology*
D017304:Annexin A5; D000900:Anti-Bacterial Agents; D005456:Fluorescent Dyes; D010718:Phosphatidylserines; D000001:Calcimycin; D054497:Phospholipases A2, Secretory; D010196:Pancreatic Elastase; D004996:Ethidium