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Fig. 2. NO mobilises stored Ca2+ in sperm. (A) Spermine
NONOate causes a slowly developing rise in [Ca2+]i in
human sperm. Responses of four separate cells are shown. Red trace shows
example of a cell generating [Ca2+]i oscillations.
(B) In low-Ca2+ medium ([Ca2+]
5 µM), the
response to NONOate was similar, but oscillations were rarely seen. Responses
of seven cells are shown. (C) Pseudocolour image series showing
NONOate-induced rise in [Ca2+]i in the sperm
neck/midpiece. Numbers show minutes since application of 100 µM spermine
NONOate. (D) Mean normalised increase in fluorescence 10 minutes after
application of 100 µM spermine NONOate to cells bathed in sEBSS (271 cells;
three experiments) and low-Ca2+ sEBSS (214 cells; three
experiments). (E) A rapid decrease in [Ca2+]i
followed washout of NONOate, followed by slow recovery. Upon re-introduction
of NONOate, many cells generated oscillations in the neck/midpiece region.
Responses of five individual cells shown. Lower panel shows pseudocolour
images series of a single [Ca2+]i oscillation (numbers
show time in seconds).
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