Potato aspartic proteases (StAPs) exert cytotoxic activity on bovine and human spermatozoa.
OBJECTIVE
To evaluate the in vitro spermicidal activity of Solanum tuberosum aspartic proteinases (StAPs) on bovine and human sperm.


METHODS
Controlled laboratory study.


METHODS
Three research laboratories at a university of biologic science. ANIMAL(S) AND DONOR(S): Frozen semen from five Aberdeen Angus bulls and six proven fertile men volunteers.


METHODS
The effect of StAPs on sperm motility was studied in vitro by incubation of different concentrations of StAPs with sperm suspensions, and motility was assessed by direct microscopic observation. Membrane integrity was analyzed by SYTOX Green uptake after incubation with different StAP concentrations. The effect of StAPs was evaluated by human erythrocyte lysis, as a control in somatic cells. The StAPs binding was monitored by fluorescence.


METHODS
Total and progressive sperm motility; hypoosmotic swelling test and SYTOX Green uptake as a measure of membrane damage; fluorescein isothiocyanate-labeled StAP binding by an optical microscopy.


RESULTS
The StAPs reduced sperm motility in a dose-dependent manner, and 25 microM of StAP1 and 35 microM of StAP3 completely abolished the progressive motility. The StAPs were able to bind in the postacrosomal and midpiece region only in bovine sperm. Also, StAPs caused spermatozoa agglutination. In vitro cell toxicity was observed by a dose-dependent increase in hypoosmotic swelling negative sperm and SYTOX Green uptake in both human and bovine spermatozoa; however, no toxic effect was observed on erythrocytes.


CONCLUSIONS
The spermicidal effect of StAPs involves plasma membrane permeabilization.
Cesari
Andreina
A
Laboratory of Biochemistry and Molecular Biology of Microorganism and Spermatozoa, Instituto de Investigaciones Biológicas, Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Mar del Plata, Buenos Aires, Argentina. acesari@mdp.edu.ar
Falcinelli
Andrea L
AL
Mendieta
Julieta R
JR
Pagano
Mariana R
MR
Mucci
Nicolas
N
Daleo
Gustavo R
GR
Guevara
María G
MG
D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000302:isolation & purification / Q000633:toxicity*; D002417:Cattle; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D003603:Cytotoxins / Q000302:isolation & purification / Q000633:toxicity*; D006801:Humans; D008297:Male; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity; D011198:Solanum tuberosum / Q000201:enzymology*; D013081:Sperm Motility / Q000187:drug effects / Q000502:physiology; D013089:Spermatocidal Agents / Q000302:isolation & purification / Q000633:toxicity; D013094:Spermatozoa / Q000187:drug effects* / Q000502:physiology
D003603:Cytotoxins; D010936:Plant Extracts; D013089:Spermatocidal Agents; D016282:Aspartic Acid Endopeptidases