Measurement of intracellular calcium levels of human spermatozoa acrosome reacted by electropermeabilization.
Human spermatozoa readily undergo in vitro capacitation and the acrosome reaction when exposed to a single, brief, high-voltage electric pulse in a medium containing calcium. Using the fluorescent calcium indicator, Fura-2, the changes in the intracellular calcium concentration of human sperm following exposure to voltages in the range of 200-2000 V cm-1 were measured. The induction of the acrosome reaction by the electropermeabilization pulse was also monitored. In comparison, changes in calcium uptake during a standard capacitation procedure in which sperm samples were incubated for 7 h were analyzed. At hourly intervals, acrosome reaction induction and intracellular calcium uptake were measured. The results indicate that electropermeabilization rapidly and efficiently produces high populations of acrosome-reacted sperm and a corresponding and associated increase in the intracellular calcium concentration.
Wall
M
M
Department of Microbiology, University College, Galway, Ireland.
O'Flaherty
V
V
Ginty
F
F
Houghton
J A
JA
D000177:Acrosome / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002463:Cell Membrane Permeability / Q000502:physiology*; D004558:Electric Stimulation; D004951:Esterification; D005456:Fluorescent Dyes; D016257:Fura-2 / Q000031:analogs & derivatives; D006801:Humans; D008297:Male
D005456:Fluorescent Dyes; C049925:fura-2-am; D002118:Calcium; D016257:Fura-2