Hyperactivation is an important phenomenon exhibited by mammalian sperm during the process of acquiring fertilization capacity. The majority of studies have focused on incubation-induced hyperactivation in non-human species, which typically differ in size, shape, and are more homogeneous than human sperm. We develop an alternative approach via drug-induction, using high-speed imaging and analysis of same-cell changes in the flagellar movement of adhered cells. Following stimulation with 4-aminopyridine, approximately two-thirds (21 of 34) of the cells analysed exhibited a waveform with a single characteristic frequency; in all cases, the frequency was lower than before stimulation. The remaining cells (13 of 34) exhibited a more complex motility with multiple-frequency modes. The lowest mode in all cases was lower than the frequency prior to stimulation. Flagellar bending increased in all cells following stimulation and was significantly greater in the multiple-frequency responders. Despite the increased bending, time-averaged hydrodynamic power dissipation decreased significantly when assessed across all cells, the effect being significantly greater in the multiple-frequency responders than single frequency. These results reveal the heterogeneity of responses of human sperm to a hyperactivating stimulus, the methodology being potentially useful for assessing dynamic responses to stimuli in human sperm, and physiological selection of cells for assisted reproduction.
- High-speed imaging
- Sperm motility