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Articles by Ruben Peralta-Arias in JoVE
Görüntüleme Ca Teknikleri 2 + İnsan Sperm sinyal
Katherine Nash1, Linda Lefievre2, Ruben Peralta-Arias1, Jennifer Morris1, Aduen Morales-Garcia1, Tom Connolly2, Sarah Costello1, Jackson C. Kirkman-Brown3, Stephen J. Publicover1
1School of Biosciences, University of Birmingham, 2School of Medicine, University of Birmingham, 3Centre for Human Reproductive Science, Birmingham Women’s Hospital
Other articles by Ruben Peralta-Arias on PubMed
Zeitschrift Für Naturforschung. C, Journal of Biosciences. Sep-Oct, 2009 | Pubmed ID: 19957446
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.