Biochemical identification of dynein-ATPase activity in human sperm.
Dynein-ATPase is the intracellular motor for sperm motility. In the present work we assayed the dynein-ATPase activity in an axoneme-containing fraction of human sperm, free of plasma membranes, in normozoospermic and asthenozoospermic donors. Axoneme-containing fractions were isolated from semen samples obtained from healthy donors with either normozoospermia or asthenozoospermia, as indicated by a sperm motility lower than 50% (WHO grade a + b). The dynein-ATPase activity was assayed and partially characterized. The dynein-ATPase activity in the axoneme-containing fractions was identified as Mg2+-dependent ATPase activity inhibited by 10 microM vanadate. This inhibition was not seen when the assay was done in the presence of 1 mM norepinephrine. The dynein-ATPase activity is Mg2+-dependent, Li+-sensitive, and insensitive to 2 mM ouabain, 1 microM oligomycin, and 1 microM thapsigargin. The dynein-ATPase activity was significantly lower (p < 0.001) for asthenozoospermic donors as compared to normozoospermic donors. This is a straightforward dynein-ATPase assay that can be used to obtain data of functional interest in clinical or experimental settings.
Vívenes
Carmen Y
CY
Laboratorio de Bioenergética Celular, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Venezuela.
Peralta-Arias
Rubén D
RD
Camejo
María Isabel
MI
Guerrero
Kenia
K
Fernández
Víctor H
VH
Piñero
Sandy
S
Proverbio
Teresa
T
Proverbio
Fulgencio
F
Marín
Reinaldo
R
D053627:Asthenozoospermia / Q000201:enzymology; D016022:Case-Control Studies; D004398:Dyneins / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D013081:Sperm Motility; D013094:Spermatozoa / Q000201:enzymology*
D004398:Dyneins