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Fig. 3. Kinetics of induced and non-induced AR, and its relationship to cell death and motility. (Ai-iv) Single-sperm response graphs showing the timing of A23187 (10 µM)-induced AR (uptake of fluorescent anti-CD46; green lines) and cell death (uptake of PI; red lines). Each graph represents a different cell. (B) Image series of sperm stimulated with 10 µM A23187. (Ci-iv) Single-sperm response graphs show the timings of AR and cell death occurring spontaneously; (D) corresponding image series showing a single sperm undergoing spontaneous AR. (E) Single-sperm response graphs showing the timings of progesterone (3 µM)-induced AR and cell death; (F) corresponding image series showing a single sperm undergoing progesterone (3 µM)-induced AR. On all graphs, the x-axis is time relative to initiation of AR for that individual cell. Traces are normalised to 100% increase in fluorescence for each probe. Arrows on the single-cell graphs show the time at which that cell became immotile. In all image series, uptake of green fluorescence represents sperm undergoing AR (uptake of fluorescent anti-CD46) and uptake of red fluorescence represents sperm undergoing cell death (uptake of PI). Numbers on the image series indicate time in minutes. Scale bars: 5 µm.
