Generation of ROS

It is very important first to verify the triggering effects of exogenously added ROS and inhibition due to ROS scavengers and then to make sure that more than one capacitation agent is tested (see above) to ascertain that the involvement of ROS is a general phenomenon and not related to only one condition [46]. As stated above, any of the ROS mentioned induces capacitation when added exogenously and this seems to suggest a nonspecific effect of ROS. On the other hand, specific scavengers (SOD, catalase) or NOS inhibitors all block capacitation, which rather indicates that each of the ROS acts on defined and specific targets and this is much more in line with the known high level of organization cells possess. This led us to the hypothesis that capacitating spermatozoa generate ROS at extremely low levels and in the close proximity to their targets so to achieve a defined effect before these ROS spontaneously degrade or react with other cellular components. Therefore, the next challenge was the measurement of ROS in capacitating spermatozoa.

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